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Pancreatic tumor cells express cannabinoid receptors. A, total RNA was isolated from the corresponding cell lines and reverse transcription-PCR (RT-PCR) using selective primers for human CB 1 , CB 2 , or GAPDH was done. Representative of two experiments. P, Panc1; M, MiaPaCa2; B, BxPc3; C, Capan2. B, cell lysates were obtained from the corresponding cell lines and CB 1 , CB 2 , and a -tubulin protein levels were determined by Western blot. Spleen ( Sp ) was used as a positive control for CB 1 and CB 2 expression. Representative of two experiments. C, representative (63 Â ) images of CB receptor–stained 

Pancreatic tumor cells express cannabinoid receptors. A, total RNA was isolated from the corresponding cell lines and reverse transcription-PCR (RT-PCR) using selective primers for human CB 1 , CB 2 , or GAPDH was done. Representative of two experiments. P, Panc1; M, MiaPaCa2; B, BxPc3; C, Capan2. B, cell lysates were obtained from the corresponding cell lines and CB 1 , CB 2 , and a -tubulin protein levels were determined by Western blot. Spleen ( Sp ) was used as a positive control for CB 1 and CB 2 expression. Representative of two experiments. C, representative (63 Â ) images of CB receptor–stained 

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Pancreatic adenocarcinomas are among the most malignant forms of cancer and, therefore, it is of especial interest to set new strategies aimed at improving the prognostic of this deadly disease. The present study was undertaken to investigate the action of cannabinoids, a new family of potential antitumoral agents, in pancreatic cancer. We show tha...

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... of pancreatic tumor cells in vitro. We investigated the effect of cannabinoids on pancreatic tumor cells. First, we determined the expression of cannabinoid receptors in four different human pancreatic tumor cell lines as well as in biopsies of human pancreatic tumors. CB 1 and CB 2 cannabinoid receptor mRNA ( Fig. 1 A ) and protein (Fig. 1 B and C ) were expressed in Panc1, MiaPaCa2, Capan2, and BxPc3 cell lines. In addition, mRNA for cannabinoid receptors was expressed in several human pancreatic tumor biopsies, whereas in samples obtained from normal pancreatic tissue, mRNA levels for these receptors were very low or could not be detected (Fig. 1 D ). This difference between tumor and nontransformed pancreatic tissue was further con- firmed by immunofluorescence analysis of CB receptors both in human biopsies from pancreatic cancer (Fig. 1 E ) and in pancreatic tumors generated in mice (Supplementary Fig. S1; see below). In both cases, expression of cannabinoid receptors was detected clearly in the tumor nodules but hardly in the surrounding pancreatic tissue. Next, we tested the effect of THC on cell viability. Incubation with THC led to a dose-dependent decrease in cell viability in the four lines tested (Fig. 2 A ). Because cells exhibited a different sensitivity to THC treatment, we chose MiaPaCa2 as the most sensitive line and Panc1 as a less sensitive line to confirm the involvement of cannabinoid receptors in THC antiproliferative action. Incubation with the CB 2 -selective antagonist SR144528, but not with the CB 1 -selective antagonist SR141716, prevented THC- induced loss of cell viability in both lines (Fig. 2 B and C ). Likewise, in MiaPaca2 and Panc1 cells, THC led to caspase-3 activation, a characteristic of apoptotic cell death, and preincubation with SR144528 abrogated this effect (Fig. 2 D and E ). Because de novo synthesized ceramide has been implicated in CB receptor–mediated apoptosis of glioma cells (19, 20), we tested the involvement of this pathway in our model. As shown in Supplementary Fig. S2 A , incubation with THC led to ceramide accumulation in MiaPaca2 and Panc1 cell lines, and preincubation with SR144528 or ISP-1, a selective inhibitor of serine palmitoyltransferase (the enzyme that catalyzes the rate-limiting step of ceramide biosynthesis; ref. 21), prevented this accumulation. In addition, ISP-1 prevented the THC-induced ( a ) decrease of cell viability (Fig. 2 B and C ) and ( b ) activation of caspase 3 (Supplementary Fig. S2 B and C ), indicating that de novo synthesized ceramide is involved in THC-induced apoptosis of pancreatic tumor cells. apoptosis of pancreatic tumor cells. p8 (also designated as candidate of metastasis 1) is a stress-regulated protein related to the architectural factor HMG-I/Y (22). This protein has been implicated in a number of functions including the induction of apoptosis of pancreatic tumor cells (23). In addition, it has been shown that ceramide treatment leads to p8 up-regulation (24) and we have very recently identified this protein as an essential mediator of cannabinoid antitumoral action in gliomas (25). We therefore tested the involvement of this protein in the antiproliferative effect of THC in our cells. p8 mRNA levels increased after THC treatment of MiaPaca2 cells, and incubation with SR144528 (Fig. 3 A ) or ISP-1 (Fig. 3 B ) prevented this effect. Moreover, knockdown of p8 mRNA using a selective siRNA prevented THC-induced apoptosis of MiaPaCa2 cells (Fig. 3 C ), confirming the implication of this gene in the response to THC in our model. Our next step was to identify genes downstream of p8 that could participate in the antitumoral effect of THC. By comparing the mRNA expression profiles of p8-deficient fibroblasts and fibroblasts with enforced p8 expression, several p8-dependent genes have been implicated in apoptotic signaling (25). Among these genes, we selected the endoplasmic reticulum (ER) stress–related proteins ATF-4 (26, 27) and TRB3 (28) as potential mediators of p8- dependent effects in our cells. Incubation with THC led to a parallel increase in p8, ATF-4, and TRB3 mRNA levels, which was prevented by incubation with ISP-1 (Fig. 3 D ). In addition, knockdown of p8 mRNA prevented ATF-4 and TRB3 up-regulation (Fig. 3 E ). Moreover, knockdown of ATF-4 or TRB3 mRNA also prevented THC-induced apoptosis (Fig. 3 F ). Taken together, these observations support that challenge with THC triggers a ceramide- and p8-controlled apoptotic response in pancreatic tumor cells, which involves up-regulation of these genes. models in vivo . To evaluate the antiproliferative effect of cannabinoids on pancreatic tumors in vivo , we first generated tumor xenografts by s.c. injection of MiaPaCa2 cells in immunodeficient mice. As shown in Fig. 4, peritumoral treatment with THC or the CB 2 -selective (and therefore psychoactivity devoid; ref. 19) cannabinoid agonist JWH-133 reduced notably the growth of the established pancreatic tumors. Next, we generated tumors by intrapancreatic injection of MiaPaCa2 cells to investigate the antitumoral effect of cannabinoids in a model that resembles more closely the niche of pancreatic cancer spreading. The synthetic cannabinoid agonist WIN 55,212-2 was selected for i.p. administration owing to its better bioavailability than THC and other classic cannabinoids. Experiments were done to confirm that this compound, which exhibits affinity for both cannabinoid receptor types (7), induces apoptosis of pancreatic cancer cells via the same endoplasmic reticulum stress–related proapoptotic pathway as THC (Supplementary Fig. S3). Administration of ...
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... of pancreatic tumor cells in vitro. We investigated the effect of cannabinoids on pancreatic tumor cells. First, we determined the expression of cannabinoid receptors in four different human pancreatic tumor cell lines as well as in biopsies of human pancreatic tumors. CB 1 and CB 2 cannabinoid receptor mRNA ( Fig. 1 A ) and protein (Fig. 1 B and C ) were expressed in Panc1, MiaPaCa2, Capan2, and BxPc3 cell lines. In addition, mRNA for cannabinoid receptors was expressed in several human pancreatic tumor biopsies, whereas in samples obtained from normal pancreatic tissue, mRNA levels for these receptors were very low or could not be detected (Fig. 1 D ). This difference between tumor and nontransformed pancreatic tissue was further con- firmed by immunofluorescence analysis of CB receptors both in human biopsies from pancreatic cancer (Fig. 1 E ) and in pancreatic tumors generated in mice (Supplementary Fig. S1; see below). In both cases, expression of cannabinoid receptors was detected clearly in the tumor nodules but hardly in the surrounding pancreatic tissue. Next, we tested the effect of THC on cell viability. Incubation with THC led to a dose-dependent decrease in cell viability in the four lines tested (Fig. 2 A ). Because cells exhibited a different sensitivity to THC treatment, we chose MiaPaCa2 as the most sensitive line and Panc1 as a less sensitive line to confirm the involvement of cannabinoid receptors in THC antiproliferative action. Incubation with the CB 2 -selective antagonist SR144528, but not with the CB 1 -selective antagonist SR141716, prevented THC- induced loss of cell viability in both lines (Fig. 2 B and C ). Likewise, in MiaPaca2 and Panc1 cells, THC led to caspase-3 activation, a characteristic of apoptotic cell death, and preincubation with SR144528 abrogated this effect (Fig. 2 D and E ). Because de novo synthesized ceramide has been implicated in CB receptor–mediated apoptosis of glioma cells (19, 20), we tested the involvement of this pathway in our model. As shown in Supplementary Fig. S2 A , incubation with THC led to ceramide accumulation in MiaPaca2 and Panc1 cell lines, and preincubation with SR144528 or ISP-1, a selective inhibitor of serine palmitoyltransferase (the enzyme that catalyzes the rate-limiting step of ceramide biosynthesis; ref. 21), prevented this accumulation. In addition, ISP-1 prevented the THC-induced ( a ) decrease of cell viability (Fig. 2 B and C ) and ( b ) activation of caspase 3 (Supplementary Fig. S2 B and C ), indicating that de novo synthesized ceramide is involved in THC-induced apoptosis of pancreatic tumor cells. apoptosis of pancreatic tumor cells. p8 (also designated as candidate of metastasis 1) is a stress-regulated protein related to the architectural factor HMG-I/Y (22). This protein has been implicated in a number of functions including the induction of apoptosis of pancreatic tumor cells (23). In addition, it has been shown that ceramide treatment leads to p8 up-regulation (24) and we have very recently identified this protein as an essential mediator of cannabinoid antitumoral action in gliomas (25). We therefore tested the involvement of this protein in the antiproliferative effect of THC in our cells. p8 mRNA levels increased after THC treatment of MiaPaca2 cells, and incubation with SR144528 (Fig. 3 A ) or ISP-1 (Fig. 3 B ) prevented this effect. Moreover, knockdown of p8 mRNA using a selective siRNA prevented THC-induced apoptosis of MiaPaCa2 cells (Fig. 3 C ), confirming the implication of this gene in the response to THC in our model. Our next step was to identify genes downstream of p8 that could participate in the antitumoral effect of THC. By comparing the mRNA expression profiles of p8-deficient fibroblasts and fibroblasts with enforced p8 expression, several p8-dependent genes have been implicated in apoptotic signaling (25). Among these genes, we selected the endoplasmic reticulum (ER) stress–related proteins ATF-4 (26, 27) and TRB3 (28) as potential mediators of p8- dependent effects in our cells. Incubation with THC led to a parallel increase in p8, ATF-4, and TRB3 mRNA levels, which was prevented by incubation with ISP-1 (Fig. 3 D ). In addition, knockdown of p8 mRNA prevented ATF-4 and TRB3 up-regulation (Fig. 3 E ). Moreover, knockdown of ATF-4 or TRB3 mRNA also prevented THC-induced apoptosis (Fig. 3 F ). Taken together, these observations support that challenge with THC triggers a ceramide- and p8-controlled apoptotic response in pancreatic tumor cells, which involves up-regulation of these genes. models in vivo . To evaluate the antiproliferative effect of cannabinoids on pancreatic tumors in vivo , we first generated tumor xenografts by s.c. injection of MiaPaCa2 cells in immunodeficient mice. As shown in Fig. 4, peritumoral treatment with THC or the CB 2 -selective (and therefore psychoactivity devoid; ref. 19) cannabinoid agonist JWH-133 reduced notably the growth of the established pancreatic tumors. Next, we generated tumors by intrapancreatic injection of MiaPaCa2 cells to investigate the antitumoral effect of cannabinoids in a model that resembles more closely the niche of pancreatic cancer spreading. The synthetic cannabinoid agonist WIN 55,212-2 was selected for i.p. administration owing to its better ...
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... of pancreatic tumor cells in vitro. We investigated the effect of cannabinoids on pancreatic tumor cells. First, we determined the expression of cannabinoid receptors in four different human pancreatic tumor cell lines as well as in biopsies of human pancreatic tumors. CB 1 and CB 2 cannabinoid receptor mRNA ( Fig. 1 A ) and protein (Fig. 1 B and C ) were expressed in Panc1, MiaPaCa2, Capan2, and BxPc3 cell lines. In addition, mRNA for cannabinoid receptors was expressed in several human pancreatic tumor biopsies, whereas in samples obtained from normal pancreatic tissue, mRNA levels for these receptors were very low or could not be detected (Fig. 1 D ). This difference between tumor and nontransformed pancreatic tissue was further con- firmed by immunofluorescence analysis of CB receptors both in human biopsies from pancreatic cancer (Fig. 1 E ) and in pancreatic tumors generated in mice (Supplementary Fig. S1; see below). In both cases, expression of cannabinoid receptors was detected clearly in the tumor nodules but hardly in the surrounding pancreatic tissue. Next, we tested the effect of THC on cell viability. Incubation with THC led to a dose-dependent decrease in cell viability in the four lines tested (Fig. 2 A ). Because cells exhibited a different sensitivity to THC treatment, we chose MiaPaCa2 as the most sensitive line and Panc1 as a less sensitive line to confirm the involvement of cannabinoid receptors in THC antiproliferative action. Incubation with the CB 2 -selective antagonist SR144528, but not with the CB 1 -selective antagonist SR141716, prevented THC- induced loss of cell viability in both lines (Fig. 2 B and C ). Likewise, in MiaPaca2 and Panc1 cells, THC led to caspase-3 activation, a characteristic of apoptotic cell death, and preincubation with SR144528 abrogated this effect (Fig. 2 D and E ). Because de novo synthesized ceramide has been implicated in CB receptor–mediated apoptosis of glioma cells (19, 20), we tested the involvement of this pathway in our model. As shown in Supplementary Fig. S2 A , incubation with THC led to ceramide accumulation in MiaPaca2 and Panc1 cell lines, and preincubation with SR144528 or ISP-1, a selective inhibitor of serine palmitoyltransferase (the enzyme that catalyzes the rate-limiting step of ceramide biosynthesis; ref. 21), prevented this accumulation. In addition, ISP-1 prevented the THC-induced ( a ) decrease of cell viability (Fig. 2 B and C ) and ( b ) activation of caspase 3 (Supplementary Fig. S2 B and C ), indicating that de novo synthesized ceramide is involved in THC-induced apoptosis of pancreatic tumor cells. apoptosis of pancreatic tumor cells. p8 (also designated as candidate of metastasis 1) is a stress-regulated protein related to the architectural factor HMG-I/Y (22). This protein has been implicated in a number of functions including the induction of apoptosis of pancreatic tumor cells (23). In addition, it has been shown that ceramide treatment leads to p8 up-regulation (24) and we have very recently identified this protein as an essential mediator of cannabinoid antitumoral action in gliomas (25). We therefore tested the involvement of this protein in the antiproliferative effect of THC in our cells. p8 mRNA levels increased after THC treatment of MiaPaca2 cells, and incubation with SR144528 (Fig. 3 A ) or ISP-1 (Fig. 3 B ) prevented this effect. Moreover, knockdown of p8 mRNA using a selective siRNA prevented THC-induced apoptosis of MiaPaCa2 cells (Fig. 3 C ), confirming the implication of this gene in the response to THC in our model. Our next step was to identify genes downstream of p8 that could participate in the antitumoral effect of THC. By comparing the mRNA expression profiles of p8-deficient fibroblasts and fibroblasts with enforced p8 expression, several p8-dependent genes have been implicated in apoptotic signaling (25). Among these genes, we selected the endoplasmic reticulum (ER) stress–related proteins ATF-4 (26, 27) and TRB3 (28) as potential mediators of p8- dependent effects in our cells. Incubation with THC led to a parallel increase in p8, ATF-4, and TRB3 mRNA levels, which was prevented by incubation with ISP-1 (Fig. 3 D ). In addition, knockdown of p8 mRNA prevented ATF-4 and TRB3 up-regulation (Fig. 3 E ). Moreover, knockdown of ATF-4 or TRB3 mRNA also prevented THC-induced apoptosis (Fig. 3 F ). Taken together, these observations support that challenge with THC triggers a ceramide- and p8-controlled apoptotic response in pancreatic tumor cells, which involves up-regulation of these genes. models in vivo . To evaluate the antiproliferative effect of cannabinoids on pancreatic tumors in vivo , we first generated tumor xenografts by s.c. injection of MiaPaCa2 cells in immunodeficient mice. As shown in Fig. 4, peritumoral treatment with THC or the CB 2 -selective (and therefore psychoactivity devoid; ref. 19) cannabinoid agonist JWH-133 reduced notably the growth of the established pancreatic tumors. Next, we generated tumors by intrapancreatic injection of MiaPaCa2 cells to investigate the antitumoral effect of cannabinoids in a model that resembles more closely the niche of pancreatic cancer spreading. The synthetic cannabinoid agonist WIN 55,212-2 was selected for i.p. administration owing to its better bioavailability than THC ...
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... of pancreatic tumor cells in vitro. We investigated the effect of cannabinoids on pancreatic tumor cells. First, we determined the expression of cannabinoid receptors in four different human pancreatic tumor cell lines as well as in biopsies of human pancreatic tumors. CB 1 and CB 2 cannabinoid receptor mRNA ( Fig. 1 A ) and protein (Fig. 1 B and C ) were expressed in Panc1, MiaPaCa2, Capan2, and BxPc3 cell lines. In addition, mRNA for cannabinoid receptors was expressed in several human pancreatic tumor biopsies, whereas in samples obtained from normal pancreatic tissue, mRNA levels for these receptors were very low or could not be detected (Fig. 1 D ). This difference between tumor and nontransformed pancreatic tissue was further con- firmed by immunofluorescence analysis of CB receptors both in human biopsies from pancreatic cancer (Fig. 1 E ) and in pancreatic tumors generated in mice (Supplementary Fig. S1; see below). In both cases, expression of cannabinoid receptors was detected clearly in the tumor nodules but hardly in the surrounding pancreatic tissue. Next, we tested the effect of THC on cell viability. Incubation with THC led to a dose-dependent decrease in cell viability in the four lines tested (Fig. 2 A ). Because cells exhibited a different sensitivity to THC treatment, we chose MiaPaCa2 as the most sensitive line and Panc1 as a less sensitive line to confirm the involvement of cannabinoid receptors in THC antiproliferative action. Incubation with the CB 2 -selective antagonist SR144528, but not with the CB 1 -selective antagonist SR141716, prevented THC- induced loss of cell viability in both lines (Fig. 2 B and C ). Likewise, in MiaPaca2 and Panc1 cells, THC led to caspase-3 activation, a characteristic of apoptotic cell death, and preincubation with SR144528 abrogated this effect (Fig. 2 D and E ). Because de novo synthesized ceramide has been implicated in CB receptor–mediated apoptosis of glioma cells (19, 20), we tested the involvement of this pathway in our model. As shown in Supplementary Fig. S2 A , incubation with THC led to ceramide accumulation in MiaPaca2 and Panc1 cell lines, and preincubation with SR144528 or ISP-1, a selective inhibitor of serine palmitoyltransferase (the enzyme that catalyzes the rate-limiting step of ceramide biosynthesis; ref. 21), prevented this accumulation. In addition, ISP-1 prevented the THC-induced ( a ) decrease of cell viability (Fig. 2 B and C ) and ( b ) activation of caspase 3 (Supplementary Fig. S2 B and C ), indicating that de novo synthesized ceramide is involved in THC-induced apoptosis of pancreatic tumor cells. apoptosis of pancreatic tumor cells. p8 (also designated as candidate of metastasis 1) is a stress-regulated protein related to the architectural factor HMG-I/Y (22). This protein has been implicated in a number of functions including the induction of apoptosis of pancreatic tumor cells (23). In addition, it has been shown that ceramide treatment leads to p8 up-regulation (24) and we have very recently identified this protein as an essential mediator of cannabinoid antitumoral action in gliomas (25). We therefore tested the involvement of this protein in the antiproliferative effect of THC in our cells. p8 mRNA levels increased after THC treatment of MiaPaca2 cells, and incubation with SR144528 (Fig. 3 A ) or ISP-1 (Fig. 3 B ) prevented this effect. Moreover, knockdown of p8 mRNA using a selective siRNA prevented THC-induced apoptosis of MiaPaCa2 cells (Fig. 3 C ), confirming the implication of this gene in the response to THC in our model. Our next step was to identify genes downstream of p8 that could participate in the antitumoral effect of THC. By comparing the mRNA expression profiles of p8-deficient fibroblasts and fibroblasts with enforced p8 expression, several p8-dependent genes have been implicated in apoptotic signaling (25). Among these genes, we selected the endoplasmic reticulum (ER) stress–related proteins ATF-4 (26, 27) and TRB3 (28) as potential mediators of p8- dependent effects in our cells. Incubation with THC led to a parallel increase in p8, ATF-4, and TRB3 mRNA levels, which was prevented by incubation with ISP-1 (Fig. 3 D ). In addition, knockdown of p8 mRNA prevented ATF-4 and TRB3 up-regulation (Fig. 3 E ). Moreover, knockdown of ATF-4 or TRB3 mRNA also prevented THC-induced apoptosis (Fig. 3 F ). Taken together, these observations support that challenge with THC triggers a ceramide- and p8-controlled apoptotic response in pancreatic tumor cells, which involves up-regulation of these genes. models in vivo . To evaluate the antiproliferative effect of cannabinoids on pancreatic tumors in vivo , we first generated tumor xenografts by s.c. injection of MiaPaCa2 cells in immunodeficient mice. As shown in Fig. 4, peritumoral treatment with THC or the CB 2 -selective (and therefore psychoactivity devoid; ref. 19) cannabinoid agonist JWH-133 reduced notably the growth of the established pancreatic tumors. Next, we generated tumors by intrapancreatic injection of MiaPaCa2 cells to investigate the antitumoral effect of cannabinoids in a model that resembles more closely the niche of pancreatic cancer spreading. The synthetic cannabinoid agonist WIN 55,212-2 was selected for i.p. administration owing to its better bioavailability than THC and other classic cannabinoids. Experiments were done to confirm that this compound, which exhibits affinity for both cannabinoid receptor types (7), induces apoptosis of pancreatic cancer cells via the same endoplasmic reticulum stress–related proapoptotic pathway as THC (Supplementary Fig. S3). Administration of WIN 55,212-2 not only remarkably reduced the growth of intrapancreatic tumors (Fig. 5 A ) but also significantly decreased the extension of the tumor cells to proximal (Fig. 5 B ) and distal (Fig. ...

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... Over 100 phytocannabinoids have been identified (Mehmedic et al. 2010), but Δ 9 -tetrahydrocannabinol (THC) and cannabidiol (CBD) are the most common cannabinoids produced in the Cannabis plant (de Meijer et al. 2003;Mechoulam 2005). Interestingly, studies have found that multiple compounds from cannabis inhibit tumor cell growth and induce apoptosis in various cancer cells (Blázquez et al. , 2008Guzmán et al. 2006;Carracedo et al. 2006;Javid et al. 2016;Blasco-Benito et al. 2018;Tomko et al. 2019), but little is known about their effects in bladder cancer. Recently, a study suggested that cannabis-derived cannabichromene (CBC) and ∆ 9 -tetrahydrocannabinol displayed some synergy when used together in a model of urothelial cell carcinoma (Anis et al. 2021). ...
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Introduction Several studies have shown anti-tumor effects of components present in cannabis in different models. Unfortunately, little is known about the potential anti-tumoral effects of most compounds present in cannabis in bladder cancer and how these compounds could potentially positively or negatively impact the actions of chemotherapeutic agents. Our study aims to evaluate the effects of a compound found in Cannabis sativa that has not been extensively studied to date, cannflavin A, in bladder cancer cell lines. We aimed to identify whether cannflavin A co-treatment with agents commonly used to treat bladder cancer, such as gemcitabine and cisplatin, is able to produce synergistic effects. We also evaluated whether co-treatment of cannflavin A with various cannabinoids could produce synergistic effects. Methods Two transitional cell carcinoma cell lines were used to assess the cytotoxic effects of the flavonoid cannflavin A up to 100 μM. We tested the potential synergistic cytotoxic effects of cannflavin A with gemcitabine (up to 100 nM), cisplatin (up to 100 μM), and cannabinoids (up to 10 μM). We also evaluated the activation of the apoptotic cascade using annexin V and whether cannflavin A has the ability to reduce invasion using a Matrigel assay. Results Cell viability of bladder cancer cell lines was affected in a concentration-dependent fashion in response to cannflavin A, and its combination with gemcitabine or cisplatin induced differential responses—from antagonistic to additive—and synergism was also observed in some instances, depending on the concentrations and drugs used. Cannflavin A also activated apoptosis via caspase 3 cleavage and was able to reduce invasion by 50%. Interestingly, cannflavin A displayed synergistic properties with other cannabinoids like Δ ⁹ -tetrahydrocannabinol, cannabidiol, cannabichromene, and cannabivarin in the bladder cancer cell lines. Discussion Our results indicate that compounds from Cannabis sativa other than cannabinoids, like the flavonoid cannflavin A, can be cytotoxic to human bladder transitional carcinoma cells and that this compound can exert synergistic effects when combined with other agents. In vivo studies will be needed to confirm the activity of cannflavin A as a potential agent for bladder cancer treatment.
... More extensive research is needed to determine the full potential of synthetic cannabinoids in cancer. The anti-cancer effects of cannabinoids have been reported in the case of pancreatic cancer (31)(32)(33)(34). It has been demonstrated that cannabidiol and tetrahydrocannabinol can suppress pancreatic cancer growth, and that this may be partially through inhibition of p-21 activated kinase 1 (PAK1) (34). ...
... Parallel to our study, it has been shown that both CB1 and CB2 receptor agonists act through a widely common mechanism that involves cell growth regulation and apoptosis in pancreatic adenocarcinoma (35). Again, it has been shown that cannabinoids induce apoptosis on pancreatic cancer cells by the activation of the p8-ATF-4-TRB3 proapoptotic pathway (31). Dando et al stated that in pancreatic adenocarcinoma cells, autophagy induction dependent to cannabinoids was related to ROS-dependent increase of the AMP/ATP ratio (32). ...
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Objective: Cancer ranks first among the causes of morbidity and mortality all over the world, and it is expected to continue to be the main cause of death in the coming years. Therefore, new molecular targets and therapeutic strategies are urgently needed. In many cases, some reports show increased levels of endocannabinoids and their receptors in cancer, a condition often associated with tumour aggressiveness. Recent studies have suggested that cannabinoid-1/2 receptors contribute to tumour growth in a variety of cancers, including pancreatic, colon, prostate, and breast cancer. Understanding how cannabinoids can regulate key cellular processes involved in tumorigenesis, such as: cell proliferation and cell death, is crucial to improving existing and new therapeutic approaches for the cancer patients. The present study was aimed to characterize the in-vitro effect of L-759633 (a selective CB2 receptor agonist), ACPA (a selective CB1 receptor agonist) and ACEA (a selective CB1 receptor agonist) on the cell proliferation, clonogenicity, and apoptosis in pancreatic (PANC1) and breast (MDA-MB-231) cancer cells. Methods: The viability and/or proliferation of cells were detected by MTS assay. A clonogenic survival assay was used to detect the ability of a single cell to grow into a colony. Apoptosis was determined with Annexin V staining (Annexin V-FITC/PI test) and by analyzing the expression of Bcl-2-associated X protein (Bax) and B-cell lymphoma 2 (Bcl-2). Results: We found that selective CB1/2 agonists suppressed cell proliferation, clonogenicity and induced proapoptotic function in human PANC1 pancreatic and MDA-MB-231 breast cancer cells. Based on our findings, these agonists led to the inhibition of both cell viability and clonogenic growth in a dose dependent manner. CB1/2 agonists were observed to induce intrinsic apoptotic pathway by upregulating Bax, while downregulating Bcl-2 expression levels. Conclusion: Our data suggests that CB1/2 agonists have the therapeutic potential through the inhibition of survival of human PANC1 pancreatic and MDA-MB-231 breast cancer cells and also might be linked with further cellular mechanisms for the prevention (Fig. 5, Ref. 49).
... The mechanism by which accumulation of the sphingolipid ceramide leads to apoptosis has been reported to be mediated by the stress-regulated protein p8, which is upregulated by ceramide accumulation. p8 upregulation leads to the upregulation of the activating transcription factor 4 (ATF-4) and the C/EBP-homologous protein and through which induce apoptosis [56]. ...
... Cannabinoids also cause cell cycle arrest in cancer cells of prostate carcinoma [56], thyroid epithelioma [57], breast carcinoma [44], lung carcinoma [51], and gastric carcinoma [58]. It has been suggested that activation of cannabinoid receptors lead to the inhibition of adenylyl cyclase and the cAMP/protein kinase A (PKA) pathway. ...
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Triple-negative breast cancer (TNBC) is a subtype of breast cancer characterized by the lack of estrogen receptors, progesterone receptors, and HER-2 receptors. Thus, TNBC tumors do not benefit from the current therapies targeting estrogen receptor or HER-2. Therefore, there is an urgent need to develop novel treatments for this subtype of breast cancer. Marijuana is a common name given to Cannabis plants, a group of plants in the Cannabis genus of the Cannabaceae family. Cannabis plants are among the oldest cultivated crops, traced back at least 12,000 years and are well known for their multipurpose usage, including medicinal purposes. The main active compounds extracted from Cannabis plants are 21-carbon-containing terpenophenolics, which are referred to as phytocannabinoids. Of these, the tetrahydrocannabinol (THC) group contains highly potent cannabinoids, including delta-9-tetrahydrocannabinol (9-THC) and delta-8-tetrahydrocannabinol(8-THC), which are the most abundant THCs and are primarily responsible for the psychological and physiological effects of marijuana. The use of Cannabis plants for medicinal purposes was first recorded in2337 BC in China, where Cannabis plants were used to treat pains, rheumatism, and gout. Recently, several cannabinoids have been approved for several treatments, one of which is the treatment of nausea and vomiting caused by chemotherapy in cancer patients. Furthermore, increasing evidence shows that cannabinoids not only attenuate side effects due to cancer treatment but might also potentially possess direct antitumor effects in several cancer types, including breast cancer. However, the antitumor activity of cannabinoids has been variable in different studies and even promoted tumor growth in some cases. In addition, mechanisms of cannabinoids actions in cancer remain unclear. This review summarizes evidence about the mixed actions and mechanisms of cannabinoids in cancer in general and TNBC in particular.
... To date, many studies using immunohistochemical staining, Western blotting, qRT-PCR, or a combined method have demonstrated overexpression or expression of CB1R and/or CB2R in human cancers, including glioma [75][76][77][78], lymphoma [82,83], leukemia [84,85], breast [64][65][66][67], lung [60,61], ovarian [86,87], pancreatic [88], prostate [89][90][91], skin [52, 92,93] and thyroid cancers [94], endometrial [95], esophageal [96], head and neck [97], hepatocellular [98][99][100], renal [101,102], and mobile tongue carcinomas [39,103]. In addition, CB1 and CB2 receptors were highly expressed in non-Hodgkin's lymphoma, and CB1 in mantle cell lymphoma compared to reactive lymph nodes [83,104]. ...
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Citation: Cherkasova, V.; Wang, B.; Gerasymchuk, M.; Fiselier, A.; Kovalchuk, O.; Kovalchuk,
... To date, many studies using immunohistochemical staining, Western blotting, qRT-PCR, or a combined method have demonstrated overexpression or expression of CB1R and/or CB2R in human cancers, including glioma [75][76][77][78], lymphoma [82,83], leukemia [84,85], breast [64][65][66][67], lung [60,61], ovarian [86,87], pancreatic [88], prostate [89][90][91], skin [52, 92,93] and thyroid cancers [94], endometrial [95], esophageal [96], head and neck [97], hepatocellular [98][99][100], renal [101,102], and mobile tongue carcinomas [39,103]. In addition, CB1 and CB2 receptors were highly expressed in non-Hodgkin's lymphoma, and CB1 in mantle cell lymphoma compared to reactive lymph nodes [83,104]. ...
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The endocannabinoid system (ECS) is an ancient homeostasis mechanism operating from embryonic stages to adulthood. It controls the growth and development of many cells and cell lineages. Dysregulation of the components of the ECS may result in uncontrolled proliferation, adhesion, invasion, inhibition of apoptosis and increased vascularization, leading to the development of various malignancies. Cancer is the disease of uncontrolled cell division. In this review, we will discuss whether the changes to the ECS are a cause or a consequence of malignization and whether different tissues react differently to changes in the ECS. We will discuss the potential use of cannabinoids for treatment of cancer, focusing on primary outcome/care—tumor shrinkage and eradication, as well as secondary outcome/palliative care—improvement of life quality, including pain, appetite, sleep, and many more factors. Finally, we will complete this review with the chapter on sex- and gender-specific differences in ECS and response to cannabinoids, and equality of the access to treatments with cannabinoids.
... The administration of CBN, Δ9-THC and Δ8-THC from Cannabis plant both in vitro and in vivo demonstrated the inhibition of Lewis lung adenocarcinoma cell growth in mice. Since then, the antimetastatic, anti-angiogenic, pro-apoptotic and antiproliferative effects of cannabinoids proved their effectiveness in various types of cancer including glioma, lung, skin, lymphoma, thyroid, uterus, ovary, neuroblastoma, pancreas, prostate, colorectal, liver, and breast carcinoma with in vitro and in vivo models (Galve-Roperh et al. 2000;Maccrrone et al. 2000;Melk et al. 2000;Sánchez et al. 2001;Casanova et al. 2003;Sarfaraz et al. 2005;Blázquez et al. 2006;Caffarel et al. 2006;Carracedo et al. 2006;Carracedo et al. 2006;Cianchi et al. 2008;Preet et al. 2008;Caffarel et al. 2010;Appendino et al. 2011;Piñeiro et al. 2011;Vara et al. 2011;Aviello et al. 2012;Tariq et al. 2012;De Petrocellis et al. 2013;Borrelli et al. 2014;Milion et al. 2020 and references therein). During the literature search, no published data was available on the anticancer actions of Cannabis from Pakistan. ...
... The administration of CBN, Δ9-THC and Δ8-THC from Cannabis plant both in vitro and in vivo demonstrated the inhibition of Lewis lung adenocarcinoma cell growth in mice. Since then, the antimetastatic, anti-angiogenic, pro-apoptotic and antiproliferative effects of cannabinoids proved their effectiveness in various types of cancer including glioma, lung, skin, lymphoma, thyroid, uterus, ovary, neuroblastoma, pancreas, prostate, colorectal, liver, and breast carcinoma with in vitro and in vivo models (Galve-Roperh et al. 2000;Maccrrone et al. 2000;Melk et al. 2000;Sánchez et al. 2001;Casanova et al. 2003;Sarfaraz et al. 2005;Blázquez et al. 2006;Caffarel et al. 2006;Carracedo et al. 2006;Carracedo et al. 2006;Cianchi et al. 2008;Preet et al. 2008;Caffarel et al. 2010;Appendino et al. 2011;Piñeiro et al. 2011;Vara et al. 2011;Aviello et al. 2012;Tariq et al. 2012;De Petrocellis et al. 2013;Borrelli et al. 2014;Milion et al. 2020 and references therein). During the literature search, no published data was available on the anticancer actions of Cannabis from Pakistan. ...
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Background: Cannabis has a very extensive history of its uses as a medicinal plant that likely dates back more than two millennia. This review was envisioned to provide a brief summary on ethnobotany, phytochemistry, medicinal uses and some biological activities of Cannabis (hemp) with emphasis on its legalization and regulation in Pakistan. Methods: The data on Cannabis was assembled from International scientific databases like Google Scholar, PubMed/Medline, Researchgate, SciELO, Scopus, Science Direct, Taylor and Francis, Web of Science, books, government reports, Master’s and Ph.D. dissertations using specific keywords. Results: In more than 33 different regions of Pakistan, the folk medicinal uses of Cannabis against ~60 ailments are still continuing. Phytochemistry data showed that more than 70 different compounds were reported in Cannabis from Pakistan with potential antioxidant activity. Overall, the antimicrobial activity reviewed here showed that Cannabis extracts against ~19 bacterial and 8 fungal strains possess potential inhibitory effects. Data on anticancer activity of Cannabis worldwide showed remarkable outcomes against more than 12 different cancer types and no data was found on the anticancer activity from Pakistan. Conclusions: Conclusively, essential compounds isolated from Cannabis may exhibit different pharmacological actions and therefore support the utilization of species infusions and/or decoctions as folk traditional medicine in Pakistan. Through the legalization, revenue could be increased by exporting Cannabis based products or by exporting the raw material however, it should be complemented with extensive approaches to publicize the medicinal importance of Cannabis and appropriate policies should be developed for industrial and medicinal use.
... Inhibitory effect of cannabinoids was established in the pancreatic tumor growth post-treatment. Cannabinoid agonist WIN 55-212,2 also decreases the cell growth in pancreatic cancer cells through possibly the activation of TRB3, a proapoptotic protein (downstream protein of p8 and ATF-4) responsible for the apoptosis induced by ER stress (Carracedo et al. 2006). Thus, induction of apoptosis by CB2 agonist is hypothesized to slow the progression of pancreatic cancer in patients. ...
Chapter
The geriatric population is escalating globally, and the need for treating infectious and non-infectious diseases in elderly patients is also correspondingly increasing worldwide. In clinical trials and under doctor’s office settings, the drug dosages are generally computed on mg/kg body wt. basis in young adults and middle-aged men and women (<40 years). It is well recognized that in comparison with the younger age counterparts, the geriatric subjects are more susceptible to drug-mediated adverse reactions due to the reduced activity in cytochrome P450 coenzymes and glucuronidation/sulfation mechanisms. Since the body mass in elderly patients, especially frail elders, is markedly reduced due to sarcopenia, progressive loss in body fat, and osteoporosis, hence, drug doses based on mg/kg body wt. usually cannot be applied in this group of patients as is done in relatively young adults. Most of the physiological functions, including drug metabolizing and excretory capacity declines in the elderly, consequently cause significant alterations in the metabolic disposition as well as changes in the pharmacokinetic (PK) and pharmacodynamic (PD) parameters of administered drugs in elderly subjects as opposed to the younger individuals. Innumerable studies have shown wide differences in xenobiotic responses due to inter-individual variation, demographics, age, gender, ethnicity, and race. These differences are attributed to a wide array of factors such as pharmacogenetics variations, gastrointestinal and microbial metabolism of drugs, bioavailability, and first-step metabolism in the liver and renal excretion. Elderly individuals are one of the most vulnerable age groups to adverse drug reactions (ADRs) due to multiple comorbidities, co-medications, and declining functions of the gastrointestinal-hepatic-renal systems. Age-related debilitating conditions tend to enhance the incidence to ADRs and hospital readmissions due to cognitive impairment, inappropriate drug use, and drug-drug, drug-diet, and drug-herbal interactions. Collectively, all these situations make it highly challenging for the physicians, nurses, pharmacists, and surgeons to make drug dose adjustment decisions for the geriatric patients. Healthcare providers should always ask their patients about herbal and dietary supplements’ use and discourage concomitant ingestion of botanical products and fruit juices, especially grape fruit juice, with prescription drugs. Systematic research by various scientific groups and pharmaceutical companies has helped in the computation of drug dose adjustments and decision-making easier for drug administration in elderly and frail patients. Appropriate guidelines, equations, and formulas are available for calculating drug dosages for frail elderly patients based on serum creatinine or cystatin-C clearance or some other biomarkers. It is important that elderly patients should be enrolled in clinical trials for understanding the pharmacometabolomics and assessment of safety, efficacy, and optimal dose schedules of new drugs. The focus of this review is to address the age-related physiological, pharmacological, and toxicological changes in elderly humans as well as age-related alterations in the absorption, distribution, metabolism, and excretion (ADME) of drugs administered orally or by other routes. We will also describe the characteristics of drug molecules that influence the bioavailability, PK, PD, and potential interactions of prescription drugs or over-the-counter medications taken simultaneously with fruit juices and herbal remedies. In this review, we have selected examples of potential risks associated with the psychotherapeutic class of drugs because the antidepressant, antianxiety, and insomnia-treating medications are some of the most frequently used categories of drugs by elderly men and women.
... Inhibitory effect of cannabinoids was established in the pancreatic tumor growth post-treatment. Cannabinoid agonist WIN 55-212,2 also decreases the cell growth in pancreatic cancer cells through possibly the activation of TRB3, a proapoptotic protein (downstream protein of p8 and ATF-4) responsible for the apoptosis induced by ER stress (Carracedo et al. 2006). Thus, induction of apoptosis by CB2 agonist is hypothesized to slow the progression of pancreatic cancer in patients. ...
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Besides synthetic drugs, a wide variety of medicinal plants have been used for the prevention and management of various liver disorders. Generally, plant therapies are well tolerated due to their lesser side effects. The aims and objectives of this review are to describe the drug therapies used for treating liver disorders as well as the most commonly used hepatoprotective plant-derived bioactive ingredients and their formulations employed for treating various liver pathologies. The extensive literature review was conducted using different databases such as ScienceDirect, SciFinder Scholar, Wiley Online Library, PubMed, ResearchGate, Google Scholar and Chemical Abstracts (until March 2021). Our literature searches showed that a wide array of plant products or plant extracts have been used in the folklore and traditional remedies for the prevention and management of liver disorders. The complex chemical structures of many isolated plant ingredients such as flavonoids, polyphenols, and steroid-type compounds have been determined using sophisticated analytical techniques. While the pharmacological and toxicological activities of many plant products have been tested in animal models, their underlying mechanism of action remains unknown. In this review, we will describe the hepatoprotective actions of the following plants and their bioactive components: Allium sativum, Allium hirtifolium, Andrographis paniculata, Apium graveolens, Asparagus racemosus, Berberis vulgaris, Curcuma longa, Emblica officinalis, Glycyrrhiza glabra, Marrubium vulgare, Nigella sativa, Phyllanthus niruri, Picrorhiza kurroa, Solanum nigrum, Swertia chirayita, Taraxacum officinale, oleanolic acid, Cliv-92, ursolic acid, berberine, proanthocyanidins, naringenin, silymarin, andrographolide, glycyrrhizin, curcumin, rhein, geniposide and resveratrol. There are challenges and opportunities for understanding the mechanism of action of the phytotherapies used for curing liver diseases as well as discovering new drug molecules useful for targeting different liver ailments. Combinations of traditional herbal remedies found to be safe and effective are also suggested as a possible cost-effective therapeutic tool to be tested in future researches aiming to unveil novel options to treat liver disorders in humans. However, well-designed, randomized, placebo-controlled, multicentre trials are needed to establish the long-term safety and efficacy as well the optimal dose schedules required for treating different liver disorders in humans.
... ER stress induces the unfolded protein response, causing the well-regulated activation of intracellular signaling responses designed to restore protein homeostasis. ER stress-mediated apoptosis occurs via the PERK/elF2a/ATF4/CHOP axis and suppresses tumor development in several types of cancer (Carracedo et al., 2006;Guha et al., 2017;Hwang et al., 2020;Park et al., 2007;2010;Rozpedek et al., 2016;Zhang et al., 2018). Here, we identified that EHMT1 directly regulates CHOP expression by controlling H3K9 methylation in the CHOP promoter region (Fig. 2). ...
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Colorectal cancer (CRC) has a high mortality rate among cancers worldwide. To reduce this mortality rate, chemotherapy (5-fluorouracil, oxaliplatin, and irinotecan) or targeted therapy (bevacizumab, cetuximab, and panitumumab) has been used to treat CRC. However, due to various side effects and poor responses to CRC treatment, novel therapeutic targets for drug development are needed. In this study, we identified the overexpression of EHMT1 in CRC using RNA sequencing (RNA-seq) data derived from TCGA, and we observed that knocking down EHMT1 expression suppressed cell growth by inducing cell apoptosis in CRC cell lines. In Gene Ontology (GO) term analysis using RNA-seq data, apoptosis-related terms were enriched after EHMT1 knockdown. Moreover, we identified the CHOP gene as a direct target of EHMT1 using a ChIP (chromatin immunoprecipitation) assay with an anti-histone 3 lysine 9 dimethylation (H3K9me2) antibody. Finally, after cotransfection with siEHMT1 and siCHOP, we again confirmed that CHOP-mediated cell apoptosis was induced by EHMT1 knockdown. Our findings reveal that EHMT1 plays a key role in regulating CRC cell apoptosis, suggesting that EHMT1 may be a therapeutic target for the development of cancer inhibitors.
... THC was reported to act by few mechanisms such as through Tumor Necrosis factor alpha converting enzyme (TACE/ADAM 17), by induction of candidate of metastasis 1-Activating Transcriptor factor 4-Tribbles homologue 3 (P8-ATF4-TRIB3) pro apoptotic pathway [66,63]. In combination with CBD, THC was reported to act through induction of ROS, modulation of cell cycle, caspase activities, modulation of ERK and induction of apoptosis [70]. ...
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Schedule E1 is an important part of Drugs and Cosmetics Act (Government of India) that comprises the list of poisonous drugs from plant, animal and mineral origins to be consumed under medical supervision. Ayurveda, the world's oldest medicinal system has a list of drugs represented in schedule E1 that are used since thousands of years. This review reports the anti-cancer activities of fifteen toxic ayurvedic drugs from plant origin represented in Drugs and Cosmetics Act, 1940. The information was collected from the various authentic sources, compiled and summarised. The plant extracts, formulations, phytoconstituents and other preparations of these drugs have shown effective activities against mammary carcinoma, neuroblastoma, non-small cell lung carcinoma, lymphocytic leukaemia, colorectal adenocarcinoma, Ehrlich ascites carcinoma, prostate adenocarcinoma, glioblastoma asterocytoma and other malignancies. They have various mechanisms of action including Bax upregulation, Bcl2 downregulation, induction of cell cycle arrest at S phase, G2/M phase, inhibition of vascular endothelial growth factors, inhibition of Akt/mTOR signalling etc. Certain traditional ayurvedic preparations containing these plants are reported beneficial and the possibilities of these drugs as the alternative and adjuvant therapeutic agents in the current cancer care have been discussed. The studies suggest that these drugs could be utilised in future for the critical care of malignancies.