[Show abstract][Hide abstract] ABSTRACT: Transforming growth factor β (TGF- β ) is a multifunctional cytokine recognized as an important regulator of inflammatory responses. The effect of inositol hexaphosphate (IP6), a naturally occurring phytochemical, on the mRNA expression of TGF- β 1, TGF- β 2, TGF- β 3 and T β RI, T β RII, and T β RIII receptors stimulated with bacterial lipopolysaccharides (Escherichia coli and Salmonella typhimurium) and IL-1 β in intestinal cells Caco-2 for 3 and 12 h was investigated. Real-time qRT-PCR was used to validate mRNAs level of examined genes. Bacterial endotoxin promoted differential expression of TGF- β s and their receptors in a time-dependent manner. IL-1 β upregulated mRNA levels of all TGF- β s and receptors at both 3 h and 12 h. IP6 elicited the opposed to LPS effect by increasing downregulated transcription of the examined genes and suppressing the expression of TGF- β 1 at 12 h. IP6 counteracted the stimulatory effect of IL-1 β on TGF- β 1 and receptors expression by decreasing their mRNA levels. IP6 enhanced LPS- and IL-1 β -stimulated mRNA expression of TGF- β 2 and - β 3. Based on these studies it may be concluded that IP6 present in the intestinal milieu may exert immunoregulatory effects and chemopreventive activity on colonic epithelium under inflammatory conditions or during microbe-induced infection/inflammation by modulating the expression of genes encoding TGF- β s and their receptors at transcriptional level.
Mediators of Inflammation 01/2013; 2013:436894. · 2.42 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Matrix metalloproteinases (MMPs) have repeatedly been shown to play a very active role in extracellular matrix degradation associated with tumor invasion and metastasis. Tissue inhibitors of MMPs (TIMPs) are well-known for their ability to inhibit MMP activity thereby inhibiting malignant progression. Inositol hexaphosphate (IP6 phytic acid) has been recognized to have both preventive and therapeutic effects against various cancers including that of colon. In in vitro studies, IP6 has been demonstrated to inhibit cancer cell adhesion and migration. In the present study, the effect of IP6 on the expression of MMP and TIMP genes was evaluated in unstimulated and IL-1β-stimulated colon cancer cell line Caco-2.
Real-time QRT-PCR was used to validate the transcription level of selected MMP and TIMP genes in Caco-2 cells after treatment with 1 ng/ml of IL-1β, 2.5 mM of IP6, and both for 6, 12, and 24 h.
Stimulation of cells with IL-1β only resulted in an overexpression of MMP and their TIMP mRNAs. A significant decrease in MMP-13, MMP-3, MMP-2, and TIMP-1 basal expression was achieved by IP6. IP6 was also an efficient downregulator of MMP-1, MMP-9, and TIMP-2 genes transcription stimulated by IL-1β in 6 h lasting culture. After 12 h, IL-1β-induced MMP-2 mRNA expression was significantly reduced by IP6.
Proinflammatory cytokine IL-1β upregulates MMP and TIMP mRNAs expression in colon cancer epithelial cells Caco-2. IP6 (2.5 mM) influences constitutive expression of both MMP and TIMP genes and downregulates IL-1β stimulated transcription of some of these genes. IP6 exerts its anti-metastatic activity through modulation of MMP and TIMP genes expression to prevent cancer cell migration and invasion.
International Journal of Colorectal Disease 03/2012; 27(11):1419-28. · 2.24 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Quantification of p65, p50 and IκBα mRNAs was performed by real time QRT-PCR in Caco-2 cells treated with 10, 50, and 100 μg/mL of Desulfovibrio desulfuricans LPS for 1, 6, 12, and 24 h. A strong increase in expression of p65 and IλBα genes was induced by 10 and 100 μg/mL of LPS at 1 h; after 6 h higher transcript amounts of both genes were observed at 100 μg/mL LPS. The p65 expression level was significantly increased by 50 and 100 μg/mL at 12 h and lowered by all LPS doses at 24 h. No significant differences between IκBα mRNA quantity in cells exposed to LPS at 12 and 24 h were observed. No changes in expression of p50 mRNA were induced by LPS. The expression of p65 gene positively correlated with IκBα gene expression. D. desulfuricans LPS is capable of modulating transcriptional activity of p65 and IκBα genes in intestinal epithelial cells.
[Show abstract][Hide abstract] ABSTRACT: Matrix metalloproteinases 2 (MMP-2) and 9 (MMP-9) belong to a zinc dependent family of enzymes that degrade components of extracellular matrix. One postulated mechanism by which inositol hexaphosphate (phytic acid, IP6), an ubiquitous plant component, prevents the activation of MMPs may be due to its ability to chelate minerals. The aim of the study was to evaluate the expression profile of MMP-2, MMP-9 and their tissue inhibitors TIMP-1 and TIMP-2 at the mRNA level in human colorectal cancer cell line Caco-2 treated with IP6. A kinetic study of MMP-2, MMP-9 and TIMP-1, TIMP-2 mRNAs was performed after cells treatment with 1; 2.5; 5 mM IP6 for 1, 6, 12 and 24 h. Quantification of genes expression was carried out using real time QRT-PCR technique. The gene encoding MMP-9 was neither constitutively expressed nor induced by IP6 in Caco-2 cells. IP6 at the concentration of 1 mM evoked increase in MMP-2 transcript level, however, its higher doses (2.5; 5 mM) caused a decrease in this gene expression at 1 h incubation. In 24 h lasting culture along with increasing IP6 concentration, the cells expressed lower and lower MMP-2 mRNA level. In response to 1 and 2.5 mM at 6 h, the cells demonstrated an increased transcriptional activity of the TIMP-2 gene which was accompanied by a decrease in TIMP-1 gene transcription. Treatment of cells with 2.5 mM IP6 at 12 h resulted in a strong increase in both TIMP-1 and TIMP-2 expression. The results of this study show that IP6 modulates MMP-2, TIMP-1 and TIMP-2 genes expression in colon cancer cells at the transcriptional level in a way dependent on its concentration and time of interaction.
Acta poloniae pharmaceutica 01/2010; 67(6):625-9. · 0.69 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Familial hypertrophic cardiomyopathy (FHCM) is characterized by an autosomal dominant transmission, left ventricular hypertrophy and myocardial disorganization. So far, 13 genetic loci and more than 130 mutations in ten different genes have been identified. Recent study suggested impaired force production associated with inefficient use of ATP as the main disease mechanism. We performed haplotype analysis with the use of microsatellite markers linked with beta-myosin heavy chain, troponin T, alpha-tropomyosin and cardiac myosin protein C genes in three Polish families with hypertrophic cardiomyopathy (23 individuals). This method is based on the analysis of distribution of the disease in the family and the alleles of chosen microsatellite markers. In two families, the disease was associated with beta-myosin heavy chain gene. We also found a genetic carrier of the mutated gene among children of the patients. In one family the connection of the disease with the mutation in alpha-tropomyosin gene was confirmed, no sudden cardiac deaths were recorded and the degree of myocardial hypertrophy was small.
Acta poloniae pharmaceutica 01/2010; 67(6):669-72. · 0.69 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The pro-inflammatory effects of kinins are mediated by two bradykinin receptors: BR1 and BR2. The aim of this study was to evaluate the expression profile of kinin receptor genes by an estimation of mRNA levels in human nasal polyps (NP) and normal mucosa (NM).
BR1 and BR2-dependent genes differentially transcribed in NP were investigated using oligonucleotide microarray technology. The mRNA copy number of BR1, BR2 and TIMP1 genes was assessed by QRT-PCR. Thirty six eosinophilic (ENP), 17 neutrophilic nasal polyps (NNP) and 28 NM samples were included into the study.
Among 92 genes encoding proteins involved in signal transduction via B1 and B2 kinin receptors TIMP1 was found to be 2,63-fold higher in the NP than in NM. Increased TIMP1 gene expression was proved by QRT-PCR (p=0,003). Moreover two genes: FOS and PTGS1 presented higher (3,82- and 4,27-fold, respectively) expression in NM compared to NP tissues. In QRT-PCR analysis insignificantly higher expression of gene encoding BR1 in ENP [2564 mRNA copies/microg RNA (22-32863)] compared with NM [1426 copies mRNA (15-27995)] was found. mRNA expression for the BR2 in ENP [9872 copies mRNA (19-244832)] was insignificantly higher than in NM [5753 copies (46-199658)]. BR2 mRNA was the predominant transcript in most NP and NM samples followed by BR1 mRNA (p<0,01). There was a positive correlation between the expression of BR1 and BR2 in the ENP (r=0,91; p<0,01) and NNP (r=0,6; p<0,01).
We did not document any changes in the expression profile of kinin receptors in the analyzed groups, which may suggest that kinin receptors do not make an important contribution in the etiology of NP.
Advances in Medical Sciences 01/2009; 54(2):211-20. · 0.80 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The object of the study was to assess the expression of the genes encoding TNFalpha and its receptors (TNF-R1 and TNF-R2) in patients with nasal polyps (NP).
The number of the mRNA copies was assessed by QRT-PCR in RNA extracts from 16 eosinophilic (ENP) and 5 neutrophilic nasal polyps (NNP), and 9 normal mucosa (NM) samples. The expression of corresponding proteins was demonstrated using immunohistochemistry.
The mean level of mRNA copies for TNFalpha in ENP (82229c/microg) was not significantly higher when compared with controls (74869c/microg). NNP demonstrated significantly lower mean TNFalpha gene expression (7021c/microg) than the controls (p<0.05). A statistically higher mRNA TNFalpha copy number in ENP than in NNP was also revealed (p<0.01). A noticeably lower mRNA expression of TNF-R1 in ENP and NNP was seen as compared to the control group (10198c/microg vs. 30749c/microg, p<0.05 and 3440c/microg vs. 30749c/microg; p<0.05 respectively). In ENP the mean TNF-R2 mRNA copy number was markedly higher than in NNP (185c/microg vs. 7.6c/microg, p<0.05). TNF-R2 mRNA level did not differ significantly between ENP and the control group (185c/microg vs. 469c/microg). TNF-R1 expression was significantly higher than TNF-R2 at the mRNA (p<0.01) and protein (p<0.05) level both in ENP and NNP. No significant correlations in proteins expression were detected between ENP and NNP.
TNF-R1 has been identified to be a prevalent form of the TNFalpha receptor in nasal polyps which may reflect the apparent dominance of this form in TNFalpha signalling. The findings raise the possibility that the eosinophils from NP may influence biological responses through TNFalpha-dependent mechanisms. The differences between ENP and NNP relating to TNFalpha and the expression of its receptors may reflect the distinct character of those diseases.
Advances in Medical Sciences 02/2008; 53(2):263-9. · 0.96 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: TGF-beta is an important mediator of cell growth, differentiation, and proliferation and plays a significant role in both normal and pathological corneal tissue. However, the quantitative relations between TGF-beta1, -beta2 and -beta3 isoforms in human cornea still remain unclear. Therefore, the aim of this study was to determine the gene expression profile of TGF-betas in order to evaluate quantitative relations between the examined transcripts in human corneal epithelium. Transcriptional activity of TGF-beta1, 2, 3, GAPDH and beta-actin genes was estimated on the basis of mRNA copy number per 1 microg of total RNA using the real-time QRT-PCR technique with the SYBR Green I chemistry. Specificity of RT-PCR reaction was confirmed by determination of the characteristic melting temperature for each amplimer. Additionally, the RT-PCR products were separated on 6% polyacrylamide gels and visualized with silver salts. Expression of all TGF-beta genes for the corneal epithelium was determined. Comparable analysis of mRNA copies/1 mug of total RNA for each TGF-beta isoform showed that: TGF-beta1 > TGF-beta2; TGF-beta3 > TGF-beta2; TGF-beta1 = TGF-beta3 (ANOVA test P < 0.0001; post-hoc Tukey's test: TGF-beta1 and TGF-beta2, P = 0.0306; TGF-beta3 and TGF-beta2, P = 0.0045; TGF-beta1 and TGF-beta3 NS). We found different expression of the TGF-beta1, -2 and -3 isoforms in the human corneal epithelium. Such differential expression of TGF-betas suggests that each of them may play a specific role in corneal tissue.
[Show abstract][Hide abstract] ABSTRACT: Nasal polyps, according to many authors, generate as a result of chronic inflammation process with activation of cytokines, immunological reaction mediators that regulate proliferation, differentiation and cell apoptosis. Clarifying molecular mechanisms present in those disturbances may have diagnostic and prognostic value in evaluation of recurrence, dynamics and differentiation of nasal polyps as well as in their therapy.
The aim of the work was an analysis of nasal polyps on the basis of molecular, histopathological and clinical picture as well as comparing differentiated genes transcription in nasal polyps and proper nasal mucosa.
Oligonucleotide array with HGU 133A - Affymetrix were used to analyze the expression of 22,283 genes in nasal polyp tissues from 17 patients. The control group consisted of 8 tissue samples from patients after nasal septoplasty surgery.
All the samples could be classified to nasal polyps group or proper mucosa group, it reflected significant differences in genes profile expression in both groups. The evaluation of 22,283 genes transcriptions showed that in most cases nasal polyps tissue reflect classification connected with dominant inflammation cells infiltration. The data obtained let distinguish subgroups connected with clinical condition of the patients. The subgroup with massive nasal and sinus polyposis, eosinophilia and differentiated lower respiratory airways hyperactivity and the subgroup without eosinophilia infiltration may be distinguished. The data obtained suggest that molecular mechanisms may influence on the promotion and kind of inflammation process as well as the clinical course of nasal polyps.
Otolaryngologia polska. The Polish otolaryngology 01/2008; 62(3):261-6.
[Show abstract][Hide abstract] ABSTRACT: The most dangerous environmental factor for our skin condition is ultraviolet light radiation. Chronic exposition to ultraviolet light can induce epidermal atrophy, keratosis, depigmentation and dysplasia. In the dermis, UV light causes dramatic up-regulation of extracellular matrix-degrading enzymes. Matrix metalloproteinases (MMPs) are engaged in collagen, elastin and other extracellular matrix components degradation. In addition, to increase level of destructive enzymes, UV light has been shown to decrease collagen production. As a consequence of UV impact on skin, it shows signs of aging including loss of tone and elasticity, increased skin fragility, blood vessels weakness and wrinkles. The most dangerous effect of UV on skin is an increased risk of melanoma and other skin cancers. Retinoids are well known antiaging agents. For many years this vitamin has been used for the prevention and treatment of photoaging. Retinoids abolish cellular atypia, increase compacting of the stratum corneum and reduce skin hyperpigmentation caused by sun light. Recent evidence suggests that retinoids also play a role in the prevention of aging, because of its inhibitory effects on metalloproteinases expression. The aim of this study was to examine if all-trans-retinoic acid (ATRA) effects MMP-1, MMP-2, MMP-3 and MMP-14 gene expression in fibroblasts cultured in vitro.
Acta poloniae pharmaceutica 01/2008; 65(1):85-91. · 0.69 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Over the last years phytic acid, a hexaphosphorylated inositol (IP6) has attracted particular attention due to its anti-cancer activity, however, the molecular mechanisms of its action have not been elucidated, as yet. The aim of this study was to evaluate the influence of phytic acid on the expression of genes encoding p65 and p50 subunits of NF-kappaB and of its inhibitor IkappaBalpha in human colorectal cancer cell line Caco-2. A kinetic study of p65 and p50 subunits and IkappaBalpha mRNAs expression was performed on Caco-2 cells after treatment with 1, 2.5 and 5 mM IP6 for 1, 6, 12 and 24 h. Quantification of the genes expression was carried out using real time QRT-PCR technique. Treatment of cells with 5 mM IP6 resulted in a strong increase in IkappaBalpha expression at 6 h, 12 h and 24 h. The level of p65 transcript after 1 h was lower in the cells exposed to 1, 2.5, and 5 mM IP6 than in the control cells. The increase in transcriptional activity of p65 gene in response to 5 mM IP6 after 6 h and 12 h was observed. Cells treated for 24 h with 2.5 mM and 5 mM IP6 showed a significant decrease in expression of p65 gene. There were no quantitative changes in the p50 gene expression in the cells treated with IP6 compared to the control cells. High positive correlation between the expression of IkappaBalpha and p65 was detected. The results of this study suggest that IP6 primarily influences p65 and IkappaBalpha genes expression in colon cancer cells. Changes in transcriptional activities of IkappaBalpha and p65 depend on IP6 concentration and time of its action.
Acta poloniae pharmaceutica 01/2008; 65(6):697-702. · 0.69 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Recurrent polyposis in the same patient resulting in the necessity of repeated surgeries forced to search for new pharmacological therapeutic methods. At present, locally acting glycocorticosteroids have the greatest value in the treatment of nasal polyposis. Polyps grow is connected with inflammation process and proliferation of fibroblasts.
An evaluation of calcitriol and tacalcitol influence on proliferation of fibroblasts extracted from nasal polyps.
consisted of 9 tissue samples coming from nasal polyps sampled during polypectomies. The testing was performed on the polyps fibroblasts after the sixth passage after the primary culture was established. Three days after the culture was started the cells were poured with nutrient medium without serum added and after further 24 hours was replaced by nutrient medium with takalcitol and calcitriol in the defined concentrations. The expression of the genes coding histone H3 was evaluated with the use of RT-PCR technique.
Tacalcitiol and calcitriol in vitro decrease proliferation of fibroblasts sampled from nasal polyps. Inhibition is most effective for the concentration of 10-4M. Tacalcitiol and calcitriol also inhibit level of histone H3 gene expression.
Experimental data suggest tacalcitiol to be more effective in the same concentration. Present studies may indicate the direction of further investigation in the potential pharmacological treatment on nasal polyps.
Otolaryngologia polska. The Polish otolaryngology 02/2007; 61(5):661-7.
[Show abstract][Hide abstract] ABSTRACT: The formation of nasal polyps is connected with a chronic inflammatory process with the activation of different cytokines. TGF-ss induces fibrosis and acts as a chemoattractant and proliferation factor for fibroblasts. The aim of the study was to evaluate the expression profiles of the genes coding TGF-ss isoforms in nasal polyps with predominately eosinophilic and neutrophilic infiltration and in healthy mucosa and to assess their mutual correlation with the levels of gene transcription.
The study group consisted of 24 patients with nasal polyposis. On the basis of the histopathological evaluation there were 16 eosinophilic and 8 neutrophilic polyps. The control group constituted 9 healthy patients. The expression profiles of the genes coding the TGF-ss isoforms were detected using real-time RT-QPCR.
TGF-beta1 and TGF-beta2 mRNAs were revealed in 10 patients with eosinophilic polyps. TGF-beta1 transcriptional activity was accompanied by TGF-beta2 transcriptional activity in nasal polyps. TGF-beta2 gene expression in tissues without mRNA for TGF-beta1 was silenced. There was positive correlation between the expressions of the TGF-beta1 and TGF-beta2 isoforms in nasal polyps. TGF-beta1 mRNA was present at higher levels in all control samples than in eosinophilic polyps. An increased TGF-beta1 mRNA expression was accompanied by an increased TGF-beta2 mRNA expression in healthy mucosa. TGF-beta3 showed the most intensive transcriptional activity among the TGF-ss isoforms in both nasal polyps and control tissues. There was no correlation between TGF-beta3 and TGF-beta1 nor between TGF-beta3 and TGF-beta2 transcriptional activity in nasal polyps and normal tissue.
Postępy Higieny i Medycyny Doświadczalnej (Advances in Hygiene and Experimental Medicine) 02/2007; 61:702-7.
[Show abstract][Hide abstract] ABSTRACT: The objectives of the study were to estimate human cytomegalovirus (HCMV) DNA copy number in broncho-alveolar lavage cells, blood leukocytes, and serum of patients with idiopathic pulmonary fibrosis (IPF). The study groups consisted of 16 patients, newly diagnosed with IPF and never treated, (mean age 40.9 +/-11.0 yr; F/M-7/9) and in 16 adult healthy volunteers (mean age 36.8 +/-6.4 yr; F/M-4/12) used as controls. The HCMV DNA copy number was calculated by a Q-PCR method using TaqMan ABI PRISM 7700. We found that the prevalence of the HCMV DNA positive subjects in the patient group (75%) did not differ significantly from that in the control group (69%). We also found that in both patient and control groups the mean HCMV DNA copy number in BAL cells was significantly higher than that in blood leukocytes (log10=2.7 vs. 1.2 for patients and 2.8 vs. 0.9 for controls, respectively). However, a higher HCMV DNA copy number in blood serum was observed in IPF patients than in controls (log10=3.2 vs. 2.0, respectively). We conclude that the lungs play an important role in the human pathobiology of cytomegalovirus sustenance.
Journal of physiology and pharmacology: an official journal of the Polish Physiological Society 10/2004; 55 Suppl 3:67-75. · 2.48 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The aim of the study is to estimate human cytomegalovirus DNA copy number in different compartments (BAL cells, blood leukocytes, serum) in patients with idiopathic pulmonary fibrosis (IPF). There were 16 patients (mean age 40.87 +/-10.97; 9 males, 7 females) with newly diagnosed and so far not treated IPF included in the study. The diagnosis of IPF was confirmed by typical HRCT findings and/or lung biopsy histopathological examination (result of the biopsy: "usual interstitial pneumonia"). There were also 16 adult volunteers (mean age 36.75 +/- 6.43; 12 males, 4 females) included in the study as a control group. Using the real-time quantitative polymerase chain reaction the HCMV DNA copy number was estimated. The prevalence of HCMV DNA positive subjects in the IPF group (75%) was higher, but did not differ significantly from the control group (69%). The HCMV DNA copy number in 1 million BAL cells was significantly higher comparing to blood leukocytes, both in IPF and control group (log10 = 2.7 vs. 1.2 for IPF and 2.8 vs. 0.9 for control, respectively). Higher mean HCMV DNA copy number was observed in IPF patients comparing to control group (log10 = 3.2 for IPF and 2.0 for control) in 1 mL of blood serum. We concluded that the lungs play an important role in human cytomegalovirus latency and infection reactivation and thus could be a cofactor modulating the course of IPF in humans. The contribution of HCMV infection in IPF patients should be taken into account during immunosuppression therapy planning.
Pneumonologia i alergologia polska: organ Polskiego Towarzystwa Ftyzjopneumonologicznego, Polskiego Towarzystwa Alergologicznego, i Instytutu Gruzlicy i Chorob Pluc 02/2003; 71(11-12):512-20.
[Show abstract][Hide abstract] ABSTRACT: Intestinal epithelial cells play an important role in the mucosal immune and inflammatory reactions via the expression and secretion of proinflammatory cytokines such as interleukin-6 (IL-6) and interleukin-8 (IL-8). The expression of both interleukins is regulated by nuclear factor KB (NF-kappaB). Phytic acid (IP6) is an essential component of high fiber diet. It is physiologically present in the human large gut at concentrations reaching 4 mM. It exhibits pleiotropic health beneficial effects including anti-oxidant and anti-tumor activities. Recent studies showed that IP6 can modulate immune functions of intestinal epithelium through regulation of proinflammatory cytokines secretion. The aim of this study was to analyze the effect of IP6 on the expression of IL-6 and IL-8 as well as p50 and p65 subunits of NF-kappaB and its inhibitor IkappaBalpha in Caco-2 cells stimulated with IL-1beta. A kinetic study of mRNAs expression in cells was performed after their treatment with 1 and 2.5 mM IP6 for 3, 6 and 12 h. Quantification of the genes expression was carried out using real time QRT-PCR technique. IP6 at all used concentrations had no influence on transcription of p65 gene and modulated expression of p50 and IkappaBalpha genes in Caco-2 cells. Treatment of cells with IP6 resulted in a marked decrease in both IL-6 (at 3 and 6 h) and IL-8 expression (3 h). The results of these studies suggest that IP6 may exert immunoregulatory effects on intestinal epithelium by influencing transcriptional activity of genes encoding p50 subunit of NF-kappaB, its inhibitor IkappaBalpha and proinflammatory cytokines IL-6 and IL-8.
Acta poloniae pharmaceutica 69(6):1313-9. · 0.69 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Inositol hexaphosphate (IP6) is a naturally occurring phytochemical, found in abundance in cereals, legumes and other high-fiber-content diets. IP6 has shown promising efficacy against a wide range of cancers. Its anti-cancer activity involves anti-proliferative, pro-apoptotic and anti-metastatic effects. Both matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs), are implicated in tumor growth, metastasis, and angiogenesis. Phorbol-12-myristate 13-acetate (PMA) is a well-known inflammatory stimulator and tumor promoter that activates PKC and increases the invasiveness of various types of cancer cells by activating MMPs. The aim of the present study was to examine the influence of IP6 on the expression of selected MMPs, i.e., MMP-1, -2, -3, -9, 10, -13 and their TIMP-1 and -2 in unstimulated and PMA-stimulated colon cancer cell line Caco-2. Quantification of genes expression in Caco-2 cells treated with 100 ng/mL of PMA, 2.5 mM of IP6 and both for 6 and 12 h was carried out using real time QRT-PCR technique. Stimulation of cells with PMA resulted in an up-expression of MMP-2, MMP-3, MMP-9, MMP-10, MMP-13 and TIMP-1 mRNAs and decrease in MMP-1 gene expression. The quantity of TIMP-2 transcript was reduced by PMA. A significant decrease in MMP-2, MMP-3, MMP-10, MMP-13, and TIMP-1 expression in response to IP6 was observed. IP6 down-regulated MMP-9 transcription induced by PMA and decreased the level of both MMP-2 and MMP-3 mRNAs in PMA-stimulated cells. Caco-2 treated with both PMA and IP6 showed a significant decrease in MMP-1 expression in comparison to PMA-stimulated cells. The results of this study show that PMA can modulate MMP and TIMP genes transcription in colon cancer cells Caco-2. IP6 exerts an influence of basal mRNA expression of some MMPs and their tissue inhibitors and down-regulates MMP-1, MMP-2, MMP-3 and MMP-9 in cells treated with PMA. IP6 could be an effective anti-metastatic agent that suppresses expression of MMP genes at transcription level.
Acta poloniae pharmaceutica 69(6):1307-12. · 0.69 Impact Factor