Xiao Liang

Publications of Xiao Liang

• Antitumor activity of Chidamide in hepatocellular carcinoma cell lines.

  Authors: Haijuan Wang, Yi Guo, Ming Fu, Xiao Liang, Xueyan Zhang, Renzhi Wang, Chen Lin, Haili Qian

  Molecular medicine reports. 04/2012;

  Chidamide, the structural analog of MS-275, is a novel and promising histone deacetylase (HDAC) inhibitor for use in cancer therapy. To investigate its effects on cancer cell growth, MTT assay wasChidamide, the structural analog of MS-275, is a novel and promising histone deacetylase (HDAC) inhibitor for use in cancer therapy. To investigate its effects on cancer cell growth, MTT assay was performed in 10 human cancer cell lines. The data showed that the IC50 of Chidamide ranged from 1 to 13 µM, which was comparable to that of MS-275 in half of the tested cell lines. Furthermore, the growth curve indicated that cell growth was gradually inhibited with an increase in Chidamide dosage, and the inhibition was reversed after drug removal in two hepatocelluclar carcinoma cell lines, BEL-7402 and HCC-9204. To determine cell cycle and apoptosis, FACS was carried out in the BEL-7402 and HCC-9204 cells treated with Chidamide. A decrease in the cell population at S phase and an increase in the cell population at G1 phase occurred in a dose-dependent manner. In addition, Chidamide induced apoptosis and up-regulated p21 mRNA expression. These results suggest that Chidamide may arrest the cell cycle and inhibit the growth of hepatocellular carcinoma cells through up-regulation of p21. Further studies are required to clarify the antitumor activity of Chidamide in vivo and its mechanism in anticancer therapy.

• [Relationship between MTA1 expression and invasive and metastatic ability of cervical cancer cell].

  Authors: Xiao-yan Han, Hai-li Qian, Jun-jun Yang, Xue-yan Zhang, Ming Fu, Xiao Liang, Chen Lin, Yang Xiang

  Zhonghua fu chan ke za zhi. 09/2011; 46(9):678-83.

  To investigate the relationship between metastasis-associated gene 1 (MTA1) expression and invasive and metastatic ability of cervical cancer cell. Three kinds of plasmids pcDNA3 (control group),To investigate the relationship between metastasis-associated gene 1 (MTA1) expression and invasive and metastatic ability of cervical cancer cell. Three kinds of plasmids pcDNA3 (control group), pcDNA3-MTA1 (MTA1 group) and pSilencer3.1-MTA1-siRNA (MTA1-siRNA group) were transfected into human cervical cancer cell line CaSki cells. Reverse transcription (RT)-PCR and western blot were used to detected MTA1 mRNA and protein expressions. The effects of MTA1 expression on CaSki cell growth and proliferation, cell migration, adhesion and invasion, and cell cycles were tested by methyl thiazolyl tetrazolium (MTT), clone formation experiment, wound-healing assay, transwell assay, adhesion assay and flow cytometry, respectively. In animal experiment, three groups of cells were inoculated to BALB/c nude mouse subcutaneously to observe tumor formation ability. Compared with control group, MTA1 mRNA and protein were significantly overexpressed in MTA1 group, while MTA1-siRNA group showed lower MTA1 expression. Compared with control group, MTA1 group showed significantly accelerated cell growth; while MTA1-siRNA group showed decreased cell growth since the second day (P < 0.05). Clone formation number in control, MTA1 and MTA1-siRNA group were 133 ± 6, 169 ± 10 and 57 ± 5, respectively. MTA1 group showed accelerated cell formation, while MTA1-siRNA group showed the reverse effect compared with that in control group (P < 0.05). At 24, 48 and 72 hours after wounding, the healing ability of MTA1-siRNA group significantly lagged behind that in the control group, while MTA1 group showed accelerated cell healing ability. The adhesion rate of control, MTA1 and MTA1-siRNA group were (69.3 ± 3.6)%, (80.4 ± 5.6)% and (39.2 ± 7.4)% separately at 90 minutes after cell seeding. In contrast with control group, MTA1 group promoted the adhesion of CaSki cell to matrigel matrix, while MTA1-siRNA group inhibited the adhesion process (P < 0.05). In the migration assay, the number of cells migrated to the bottom side of the membrane in control, MTA1 and MTA1-siRNA group were 153 ± 17, 247 ± 38 and 82 ± 10, respectively. The number of cells in the invasion assay were 231 ± 19, 354 ± 36 and 76 ± 7, respectively. Compared with the control group, MTA1 group significantly increased the migration and invasion ability, while MTA1-siRNA group showed lower cell migration and invasion ability (P < 0.05). In cell cycle experiment, no significant differences of cell proportions including G(1), S and G(2) stage were found among three groups (P > 0.05). In animal experiment, compared with control group, MTA1 group showed accelerated tumor formation and growth, while the MTA1-siRNA group showed the reverse effect (P < 0.05). MTA1 may play its roles to promote cervical cancer cell invasion, migration, adhesion, as well as cell growth and colony formation, while RNA interference against MTA1 may decrease the malignant phenotypes. This study shows that it will be an effective beginning to explore metastasis mechanisms and cancer gene therapy strategy targeting MTA1 in cervical cancer.

• [Effect of adenovirus delivered tissue inhibitor of metalloproteinases 3 on the irradiation sensitivity of human papillomavirus positive cervical cancer cells].

  Authors: Hua-li Wei, Ying Zhang, Jun-jun Yang, Xue-yan Zhang, Xi-ting Meng, Ming Fu, Xiao Liang, Hua Duan, Chen Lin, Yang Xiang

  Zhongguo yi xue ke xue yuan xue bao. Acta Academiae Medicinae Sinicae. 10/2010; 32(5):521-5.

  To explore the effects of adenovirus-delivered tissue inhibitor of metalloprotein- ases-3 (Ad-TIMP-3) on the irradiation sensitivity of human papillomavirus (HPV)-positive cervical cancer cells. AnTo explore the effects of adenovirus-delivered tissue inhibitor of metalloprotein- ases-3 (Ad-TIMP-3) on the irradiation sensitivity of human papillomavirus (HPV)-positive cervical cancer cells. An adenovirus expressing TIMP-3 (Ad-TIMP-3), alone or in combination with irradiation,was used to treat HPV-positive cervical cancer cells HeLa-Luc and CaSki. The effects of Ad-TIMP-3 on the proliferation of HeLa-Luc and CaSki cells were detected with MTT assay. The effect of the combination of Ad-TIMP-3 and X-ray on the proliferation of cells were determined by clone formation assay. Twenty nude mice were equally randomly divided into four groups: normal control group,Ad-TIMP-3 group,X-ray group,and combination group. The size of tumor was measured separately,and tumor growth curves were drawn. Ad-TIMP-3 significantly inhibited the proliferation of HPV-positive cervical cancer cells in a dose-dependent manner. Combination of Ad-TIMP-3 and X-ray significantly decreased the clones of HeLa-Luc and CaSki than Ad-TIMP-3 or X-ray alone (P<0.05). The tumor weights were (0.216±0.098), (0.276±0.073), and (0.044±0.043) g, respectively, in Ad-TIMP-3 group, X-ray group,and combination group, which were all significantly lower than that in normal control group [(0.534±0.218) g] (all P<0.05). In addition,the tumor weight in the combination group was significantly lower than that in Ad-TIMP-3 group and X-ray group (both P<0.05). The tumor inhibition rate was 59.60%, 48.30%, and 91.80% in X-ray group, Ad-TIMP-3 group and combination group, respectively. Ad-TIMP-3 can effectively inhibit the proliferation of cervical cancer cells. When combined with X-ray,it can remarkably increase the irradiation sensitivity of HPV-positive cervical cancer cells,and thus suppress the tumorigenesis capability of these cells in vivo.

• [Relationship between the level of RRM1 expression and the sensitivity to gemcitabine in the esophageal squamous cell carcinoma cell lines.]

  Authors: Yang Luo, Chen Lin, Xue-Yan Zhang, Xiao Liang, Ming Fu, Feng-Yi Feng

  Zhonghua zhong liu za zhi [Chinese journal of oncology]. 09/2009; 31(9):660-3.

  OBJECTIVE: Ribonucleotide reductase subunit M1 (RRM1) is the intracellular target of gemcitabine (GEM). The aim of this study is to explore the relationship between the level of RRM1 expression andOBJECTIVE: Ribonucleotide reductase subunit M1 (RRM1) is the intracellular target of gemcitabine (GEM). The aim of this study is to explore the relationship between the level of RRM1 expression and the sensitivity to GEM in the esophageal squamous cell carcinoma cell lines. METHODS: Four esophageal squamous cell carcinoma cell lines (Kyse-150, Kyse-450, 9706 and Eca-109) were cultured in vitro. In the same period, RRM1 expression level was measured by RT-PCR and Western blot, and cell sensitivity to GEM was determined by CCK-8 assay. The relation between cell sensitivity and RRM1 expression was further analyzed. Kyse-450 cells were continuously cultured in the medium containing 50 nM GEM. RRM1 expression was measured at different time points to monitor the dynamic changes in the surviving cells. Inhibition of RRM1 expression by RNAi method was applied and the effect on GEM-sensitivity was further examined. RESULTS: The IC(50) of Eca-109, Kyse-150, Kyse-450 and 9706 cells were (0.92 +/- 0.17), (0.48 +/- 0.11), (0.29 +/- 0.06) and (0.02 +/- 0.01) mmol/L, respectively. The expressions of RRM1 protein and mRNA of Eca-109 cell line were the highest detected by Western blot and RT-PCR, followed by Kyse-150 and Kyse-450, and the lowest one was 9706 cell line. When Kyse-450 cells were continuously treated with 50 nmol/L GEM, the level of RRM1 protein was increasing in the surviving cells. RRM1 siRNA could effectively knock down the expression of RRM1 and significantly increase the cell sensitivity to GEM (P = 0.035). CONCLUSION: The level of RRM1 expression corelates with the cell sensitivity to gemcitabine. The cells with a lower level of RRM1 expression are more sensitive to gemcitabine.

• BH3-based Fusion Artificial Peptide Induces Apoptosis and Targets Human Colon Cancer.

  Authors: Yongjun Liu, Yunfeng Li, Haijuan Wang, Jing Yu, Hongwei Lin, Dongkui Xu, Yang Wang, Ailing Liang, Xiao Liang, Xueyan Zhang, Ming Fu, Haili Qian, Chen Lin

  Molecular therapy : the journal of the American Society of Gene Therapy. 05/2009;

  Dysregulation of apoptosis is a pilot event before cancer development and plays important roles for cancer to develop resistance to chemical therapeutics. So exploring strategies to recovery theDysregulation of apoptosis is a pilot event before cancer development and plays important roles for cancer to develop resistance to chemical therapeutics. So exploring strategies to recovery the apoptosis balance is a charming and long-endeavored aim in the attempts to conquer cancers. The present study shows an exciting potency of a fusion peptide to inhibit and target to cancer cells, which is composed of BH3 (Bcl-2 Homology 3) effector domain from PUMA (p53 upregulated modulator of apoptosis) and targeting domain of trans-activator of transcription (TAT) and DV3. The in vitro results demonstrated cancer growth inhibition by the fusion peptide in colon cancer cells, as well as in lung adenocarcinoma cell line and breast carcinoma cell line of human origin. But the viability of HEK293, a noncancerous cell line, was not affected, indicating the cancer specificity of the fusion peptide. Apoptosis activation was induced by the peptide through the mitochondria pathway. In vivo studies displayed its tumor inhibiting ability by intratumoral injection. When the fusion peptide was administered systematically by tail vein, the peptide targeted the established tumors in nude mice. No other organs were significantly involved. The fusion peptide is an artificially designed molecule worthy of further evaluation and development.Molecular Therapy (2009); doi:10.1038/mt.2009.43.

• Adenoviral-mediated gene transfer of Gadd45a results in suppression by inducing apoptosis and cell cycle arrest in pancreatic cancer cell.

  Authors: Yunfeng Li, Haili Qian, Xiao Li, Haijuan Wang, Jing Yu, Yongjun Liu, Xueyan Zhang, Xiao Liang, Ming Fu, Qimin Zhan, Chen Lin

  The journal of gene medicine. 11/2008;

  BACKGROUND: The extremely poor prognosis of patients with pancreatic ductal adenocarcinoma indicates the need for novel therapeutic approaches. The growth arrest and DNA damage-inducible (Gadd) geneBACKGROUND: The extremely poor prognosis of patients with pancreatic ductal adenocarcinoma indicates the need for novel therapeutic approaches. The growth arrest and DNA damage-inducible (Gadd) gene Gadd45a is a member of a group of genes that are induced by DNA damaging agents and growth arrest signals. METHODS: We evaluated the biological activity of Gadd45a in pancreatic ductal adenocarcinoma cancer-derived cell lines and assessed the efficacy of a combined treatment with adenoviral-mediated expression of Gadd45a (Ad-G45a) and anticancer drug (Etoposide, cisplatin, 5-fluorouracil, respectively) for the PANC1 cell line. RESULTS: Gadd45a is variously expressed in cell lines derived from pancreatic ductal adenocarcinoma cancer and adenoviral-mediated expression of Gadd45a (Ad-G45a) in these cells results in apoptosis via caspase activation and cell-cycle arrest in the G2/M phase. Gadd45a significantly increased the chemosensitivity of PANC1, which may be due to abundant apoptosis induction and cell cycle arrest. By combinational treatment of Ad-G45a infection and chemotherapeutics, Gadd45a expression was elevated to a higher extent in cancer cells with wild-type p53 than in that with knocked-out p53 status, indicating a higher chemosensitivity to cancer chemotherapy. CONCLUSIONS: Gadd45a may be a promising candidate for use in cancer gene therapy in combination with chemotherapeutic agents. Copyright (c) 2008 John Wiley & Sons, Ltd.

• Ad-PUMA sensitizes drug-resistant choriocarcinoma cells to chemotherapeutic agents.

  Authors: Yan Chen, Haili Qian, Haijuan Wang, Xueyan Zhang, Ming Fu, Xiao Liang, Yan Ma, Qimin Zhan, Chen Lin, Yang Xiang

  Gynecologic oncology. 01/2008; 107(3):505-12.

  PURPOSE/OBJECTIVE: To investigate whether exogenous PUMA expression suppresses the growth of drug-resistant choriocarcinoma cells and sensitizes them to chemotherapeutic agents. METHODS: AnPURPOSE/OBJECTIVE: To investigate whether exogenous PUMA expression suppresses the growth of drug-resistant choriocarcinoma cells and sensitizes them to chemotherapeutic agents. METHODS: An adenovirus expressing PUMA (Ad-PUMA), alone or in combination with chemotherapeutic agents (5-FU, vp16, MTX), was used to treat drug-resistant choriocarcinoma cells jeg-3/vp16 and parental jeg-3. The growth inhibitory and proapoptotic effects of Ad-PUMA both in vitro and in vivo were examined. The mechanisms of PUMA-mediated growth suppression and apoptosis were investigated by an analysis of caspase 3 activation and the change of mitochondrial membrane potential. The levels of PUMA, p53 and caspase 3 were detected by Western blotting. RESULT: PUMA was expressed lower in jeg-3/vp16 than in jeg-3. jeg-3/vp16 responded much less sensitively to 5-FU and vp16 treatment than jeg-3, though PUMA was up-regulated in both cells. Exogenous PUMA expression resulted in potent growth suppression of jeg-3/vp16 and jeg-3 through induction of apoptosis. Ad-PUMA sensitized jeg-3 and jeg-3/vp16 to chemotherapeutic agents. When Ad-PUMA 10MOI and 5-FU, vp16 or MTX were combined respectively, IC50 of drugs decreased by 8.66-, 18.66- and 13.06-fold compared with those treated by anticancer drugs alone in jeg-3/vp16, while in jeg-3, IC50 decreased only by 1.80-, 1.78- and 2.76-fold. Ad-PUMA restored the sensitivity of choriocarcinoma cells to chemotherapeutic agents by enhancing apoptosis induced by anticancer drugs. Similar results could be observed in vivo. Xenograft tumors were inhibited by Ad-PUMA or vp16. In the drug-resistant group, the inhibitory rate increased from 14.57% to 78.93% in vp16 and vp16 combined with Ad-PUMA subgroups. While in the parental group, the inhibitory rate increased but slightly, from 66.39% to 71.56%. CONCLUSION: PUMA is an important player in the therapeutic responses to chemotherapeutic agents of choriocarcinoma cells. In addition to its role in inhibiting tumor growth, low dose of Ad-PUMA significantly restored the sensitivity of choriocarcinoma cells to chemotherapeutic agents in vitro and in vivo. Exogenous PUMA is potentially useful as a sensitizer in treating drug-resistant choriocarcinoma.

• Adenovirus carrying TIMP-3: a potential tool for cervical cancer treatment.

  Authors: Ying Zhang, Haili Qian, Chen Lin, Jinghe Lang, Yang Xiang, Ming Fu, Xueyan Zhang, Xiao Liang

  Gynecologic oncology. 01/2008; 108(1):234-40.

  OBJECTIVE: Previous studies have demonstrated the pivotal roles of matrix metalloproteinases (MMPs) in cervical cancer progression. Tissue inhibitor of metalloproteinases-3 (TIMP-3) can inhibit allOBJECTIVE: Previous studies have demonstrated the pivotal roles of matrix metalloproteinases (MMPs) in cervical cancer progression. Tissue inhibitor of metalloproteinases-3 (TIMP-3) can inhibit all of MMPs, and its overexpression can retard many kinds of tumor growth. Especially, a potent bystander effect would make it to be more advisable for treating cervical cancer whose primary growth pattern is to invade adjacent structures. These characters prompted the authors to explore its further applicability in human cervical cancer. METHODS: We constructed a replication deficient adenoviral vector carrying TIMP-3 (Ad-TIMP-3). It was transferred into different cervical cancer cell lines in vitro and was injected into the tumor xenografts of nude mice in vivo. RESULTS: Overexpression of TIMP-3 by adenovirus vector arrested cervical cancer cells in G2/M phase. It also promoted growth inhibition of cancer cells by bystander effects. Ad-TIMP-3 infection produced a more potent cell killing effect than Ad-p53 (P<0.05). A synergic effect was observed when Ad-TIMP-3 and cisplatin (cDDP) were used in combination (D<1). In vivo, Ad-TIMP-3 could inhibit cervical cancer xenografts growth. Compared with Ad-TIMP-3 and cDDP groups, tumor growth in Ad-TIMP-3 combined with cDDP group was inhibited more significantly (P<0.05). CONCLUSION: These findings indicated that Ad-TIMP-3 bear a therapeutic potential for cervical cancer.

• RNA interference of metastasis-associated gene 1 inhibits metastasis of B16F10 melanoma cells in a C57BL/6 mouse model.

  Authors: Haili Qian, Jing Yu, Yunfeng Li, Haijuan Wang, Chongwen Song, Xueyan Zhang, Xiao Liang, Ming Fu, Chen Lin

  Biology of the cell / under the auspices of the European Cell Biology Organization. 11/2007; 99(10):573-81.

  BACKGROUND INFORMATION: MTA1 (metastasis-associated gene 1) has been reported to be overexpressed in cancers with high potential to metastasize. Studies of the molecular mechanisms revealed that MTA1BACKGROUND INFORMATION: MTA1 (metastasis-associated gene 1) has been reported to be overexpressed in cancers with high potential to metastasize. Studies of the molecular mechanisms revealed that MTA1 plays an important role in the process of metastasis of many types of cancer. However, the role of MTA1 in melanoma development is unclear. RESULTS: We have investigated the therapeutic value of MTA1 in the B16F10 melanoma cell line with the C57BL/6 mouse model. Studies in vitro showed that MTA1 promoted the metastatic ability of B16F10 cancer cells. MTA1 down-regulation by RNA interference greatly reversed the malignant phenotypes of cancer cells. Immunohistochemical staining of MTA1 in human melanoma samples confirmed the up-regulation of MTA1 in the process of carcinogenesis. Studies in vivo confirmed down-regulation of MTA1 suppressed the growth and experimental metastasis of B16F10 melanoma cells. CONCLUSIONS: MTA1 plays an important role in melanoma development and metastasis. It has a promising potential as a target for in cancer gene therapy or chemotherapy.

• Arsenic trioxide (As2O3) reduces the invasive and metastatic properties of cervical cancer cells in vitro and in vivo.

  Authors: Jing Yu, Haili Qian, Yunfeng Li, Yang Wang, Xueyan Zhang, Xiao Liang, Ming Fu, Chen Lin

  Gynecologic oncology. 09/2007; 106(2):400-6.

  OBJECTIVES: Arsenic trioxide (As(2)O(3)) was found to induce apoptosis in certain types of cancer cells including acute promyelocytic leukemia, and recently in solid tumors. We have previouslyOBJECTIVES: Arsenic trioxide (As(2)O(3)) was found to induce apoptosis in certain types of cancer cells including acute promyelocytic leukemia, and recently in solid tumors. We have previously demonstrated that As(2)O(3) has a therapeutic effect on cervical cancer by apoptosis promotion in vitro and in vivo. Here we further our study on the role of arsenic trioxide in regulating invasive activity of cervical cancer cells in vitro and in vivo. METHODS: The effects of As(2)O(3) on human cervical cancer cell lines (HeLa, SiHa, Caski) adhesion, migration and invasion were observed by means of cell adhesion test, cell migration test and cell invasion test. The effects of As(2)O(3) on p-IkappaB, MMP-2, E-cadherin, caveolin-1 and beta-catenin protein expressions of tumor cells were determined by Western blot. In addition, the effects of As(2)O(3) on NF-kappaB activity of tumor cells were analyzed by immunoblot in whole lysates, cytosol and nucleus, respectively. In animal experiments, cervical cancer cells TC-1 were injected into tail veins of C57BL/6 mice and then the mice were treated by intraperitoneal injection of different doses As(2)O(3). Lung weights and the foci on the surface of lungs were measured. RESULTS: As(2)O(3) inhibited attachment of tumor cells to Fibronectin and Matrigel, reduced cell motility and inhibited tumor invasion potential. As(2)O(3) treatment also resulted in a positive regulation of caveolin-1, upregulation of E-cadherin and decreased activity of beta-catenin, NF-kappaB and NF-kappaB-regulated gene MMP-2. In animal experiments, lung weights in PBS group (0.31+/-0.07 g) were significantly elevated compared with those in As(2)O(3)-treated groups (0.21+/-0.03 g and 0.17+/-0.03 g) also As(2)O(3) reduced number of metastatic lesions of lungs (15.4+/-3.5 vs. 8.3+/-2.0 and 6.3+/-2.3) in a dose-dependent manner. CONCLUSIONS: This study is the first to report the effectiveness of As(2)O(3) as an inhibitor of cervical cancer invasion both in vitro and in vivo, suggesting a potential clinical application of As(2)O(3) in cervical cancer therapies combining apoptosis induction and metastasis inhibition.

• RNA interference against metastasis-associated gene 1 inhibited metastasis of B16F10 melanoma cell in C57BL/6 model.

  Authors: Haili Qian, Jing Yu, Yunfeng Li, Haijuan Wang, Chongwen Song, Xueyan Zhang, Xiao Liang, Ming Fu, Chen Lin

  Biology of the cell / under the auspices of the European Cell Biology Organization. 06/2007;

  Background information. The metastasis-associated gene 1 (MTA1) has been reported to be overexpressed in cancers with high potential to metastasis. Studies on molecular mechanisms revealed thatBackground information. The metastasis-associated gene 1 (MTA1) has been reported to be overexpressed in cancers with high potential to metastasis. Studies on molecular mechanisms revealed that metastasis-associated gene 1 plays an important role in the process of metastasis of many cancer types. But the role of metastasis-associated gene 1 in melanoma development is unclear. Results. Here we investigated the therapeutic value of metastasis-associated gene 1 in B16F10 cell with C57BL-6 model. Biological studies in vitro showed that metastasis-associated gene 1 promoted the metastatic ability of B16F10 cancer cells. metastasis-associated gene 1 down-regulation by RNA interference (RNAi) greatly reversed the malignant phenotypes of cancer cells. Immunohistochemical staining of metastasis-associated gene 1 in human melanoma samples confirmed the up-regulation of metastasis-associated gene 1 in the process of carcinogenesis. In vivo studies confirmed down-regulation of metastasis-associated gene 1 suppressed the growth and experimental metastasis of B16F10 melanoma cells. Conclusions. Metastasis-associated gene 1 play an important role in melanoma development and metastasis. It bears a promising potential in the application targeting metastasis-associated gene 1 in cancer gene therapy or chemotherapy.

• Therapeutic effect of arsenic trioxide (As2O3) on cervical cancer in vitro and in vivo through apoptosis induction.

  Authors: Jing Yu, Haili Qian, Yunfeng Li, Yang Wang, Xueyan Zhang, Xiao Liang, Ming Fu, Chen Lin

  Cancer biology & therapy. 05/2007; 6(4):580-6.

  Arsenic trioxide (As2O3) induces apoptosis in certain types of cancer cells. But the detailed mechanisms of As2O3 efficacy are not completely known. Here we demonstrate that As2O3 has a therapeuticArsenic trioxide (As2O3) induces apoptosis in certain types of cancer cells. But the detailed mechanisms of As2O3 efficacy are not completely known. Here we demonstrate that As2O3 has a therapeutic effect on cervical cancer in vitro and in vivo. We investigated the As2O3-induced apoptosis in various cervical cancer cells. The apoptosis was triggered by mitochondrial pathway and associated with dissociation of Bcl-2 from Bax and VDAC, then the release of cytochrome c from Bax and VDAC channel, resulting in the activation of caspase-9 and caspase-3. The overexpression of Bcl-2 counteracted the As2O3-mediated apoptosis. The As2O3 treatment also resulted in an increased M phase cell cycle distribution by inducing microtubule polymerization. Two independent death-signaling pathways in cervical cancer cells were activated, one dominated by JNK/p38/GADD45 and one by p53 signals. Further investigation involving assessment of As2O3 on tumor cell growth in mice indicated that As203 also inhibited in vivo tumor growth. As2O3 as an inhibitor of cervical cancer proliferation both in vitro and in vivo suggests a potential clinical application in cervical cancer therapies.

• [Adenovirus-delivered tissue inhibitor of metalloproteinases-3 transfection increases the sensitivity of cervical cancer cells to cisplatin]

  Authors: Ying Zhang, Chen Lin, Hai-Li Qian, Jing-he Lang, Ming Fu, Xue-Yan Zhang, Xiao Liang, Hua Duan, Yang Xiang

  Zhonghua zhong liu za zhi [Chinese journal of oncology]. 02/2007; 29(1):25-9.

  OBJECTIVE: To investigate the effects of adenovirus-delivered tissue inhibitor of metalloproteinases-3 ( Ad-TIMP-3) on sensitivity of cervical cancer cells to cisplatin and evaluate the potentialOBJECTIVE: To investigate the effects of adenovirus-delivered tissue inhibitor of metalloproteinases-3 ( Ad-TIMP-3) on sensitivity of cervical cancer cells to cisplatin and evaluate the potential application of this combined scheme in cervical cancer treatment. METHODS: Cells of cervical cancer CaSKi cell line were infected with Ad-TIMP-3 in vitro. Apoptotic effect, cell cycle changes and p53 protein expression were detected. After combined treatment of those cells with cisplatin, colony formation test was performed and cytotoxicity was detected by MTT. The growth curve and tumor growth inhibition in vivo were evaluated. RESULTS: The expressions of TIMP-3 mRNA and protein were significantly upregulated after transfection. As a result, massive apoptosis was induced and the cells were arrested at G2/M phase. Exogenous overexpression of TIMP-3 increased p53 protein level markedly in spite of the backgrounds of p53 gene in cells. Combined with cisplatin treatment, the cloning efficiency was decreased. A synergism was observed by isobolic method ( D < 1 ) in vitro and tumor growth was significantly inhibited in vivo. CONCLUSION: Ad-TIMP-3 is a powerful proapoptotic agent. It increases sensitivity of the cells to cisplatin and the Ad-TIMP-3 gene therapy in combination with cisplatin could be a promising alternative in cervical cancer treatment.

• Induction of protective and therapeutic antitumour immunity using a novel tumour-associated antigen-specific DNA vaccine.

  Authors: Wenxin Sun, Haili Qian, Xueyan Zhang, Chunxia Zhou, Xiao Liang, Dongmei Wang, Ming Fu, Wenbo Ma, Shuren Zhang, Chen Lin

  Immunology and cell biology. 11/2006; 84(5):440-7.

  DNA vaccination has become an attractive immunization strategy against cancer. However, a major problem of DNA vaccination is its limited potency to be taken up by the antigen-presenting cells. InDNA vaccination has become an attractive immunization strategy against cancer. However, a major problem of DNA vaccination is its limited potency to be taken up by the antigen-presenting cells. In contrast, loss of immunogenic epitopes of tumour cells has urged the development of vaccines against multiple epitopes. In this study, we developed a novel strategy for the APC to efficiently cross-present a fusion tumour antigen, which contains both MHC class I-restricted and class II-restricted T-cell epitopes from Her-2/neu and p53 in a cognate manner. The N-terminus of the fusion Her-2/neu, p53 protein was linked to the sequence encoding for human secondary lymphoid-tissue chemokine for secretion and chemokinesis, and the C-terminus of the fusion protein was linked to a cell-binding domain of IgG (Fc portion, the cell-binding domain of IgG) for receptor-mediated internalization. Here, we show that the introduction of fused-gene DNA vaccine by gene gun reduced the size of established tumours and prolonged the lifespan of tumour-bearing mice. Results show that this DNA vaccination strategy can broadly enhance the antigen-specific cellular and humoral immune responses. This vaccine is capable of inducing adaptive immunity and may provide a novel, generic design for the development of therapeutic and preventive DNA vaccines.

• Enhanced antitumor effect against human telomerase reverse transcriptase (hTERT) by vaccination with chemotactic-hTERT gene-modified tumor cell and the combination with anti-4-1BB monoclonal antibodies.

  Authors: Xiaoyan Lin, Chunxia Zhou, Shengdian Wang, Dongmei Wang, Wenbo Ma, Xiao Liang, Chen Lin, Zheng Wang, Jie Li, Sujuan Guo, Youhui Zhang, Shuren Zhang

  International journal of cancer. Journal international du cancer. 11/2006; 119(8):1886-96.

  Human telomerase reverse transcriptase (hTERT) represents an attractive target for cancer immunotherapy because hTERT is reactivated in most human tumors. In an attempt to develop an effectiveHuman telomerase reverse transcriptase (hTERT) represents an attractive target for cancer immunotherapy because hTERT is reactivated in most human tumors. In an attempt to develop an effective vaccine against most human cancers, we constructed chemotactic-hTERT vaccine. Two hTERT fragments encoding multiple cytotoxic T lymphocyte and T helper cell epitopes were fused as a tumor antigen (named Te). The plasmid based DNA vaccine (pCCL21-Te-Fc) was constructed by linking human CCL21 and IgG Fc gene sequences to each end of Te. In poorly immunogenic B16F10 mouse melanoma model, DNA (pCCL21-Te-Fc) vaccination significantly inhibited tumor growth and all of the mice were dead by day 52. The immunization with pCCL21-Te-Fc-modified tumor cells (B16/CCL21-Te-Fc) resulted in a higher antitumor effect than DNA vaccination and 25% of tumor-bearing mice achieved long-term survival (> 120 days). The combined therapy of B16/CCL21-Te-Fc plus anti-4-1BB MAbs further enhanced the immune response, resulting in 75% of tumor-bearing mice achieved long-term survival (> 120 days) in subcutaneous model and few lung nodules in pulmonary metastasis model. Rechallenge experiment showed that a persistent memory response was successfully induced by the combined therapy. In vivo depletion of lymphocytes indicated that CD8+ T cells were essential in the antitumor activity induced by B16/CCL21-Te-Fc plus anti-4-1BB MAbs, whereas NK cells and CD4+ T cells played substantial roles. The CTL activity induced by pCCL21-Te-Fc-transfected PBMCs specifically lysed a variety of human leukocyte antigen-matched and hTERT-positive human tumor cells, suggesting pCCL21-Te-Fc could serve as a vaccine against most human cancers.

• [The feasibility and mechanism of recombinant adenovirus Ad - p 14 ARF in gene therapy of liver cancer]

  Authors: Hai-Feng Song, Hai-Li Qian, Xue-Yan Zhang, Xiao Liang, Ming Fu, Chen Lin

  Zhonghua zhong liu za zhi [Chinese journal of oncology]. 10/2006; 28(9):641-5.

  OBJECTIVE: To explore the feasibility and mechanism of recombinant adenovirus Ad-pl4ARF in cancer gene therapy. METHODS: The proliferation of different liver cancer cells was assessed by morphologyOBJECTIVE: To explore the feasibility and mechanism of recombinant adenovirus Ad-pl4ARF in cancer gene therapy. METHODS: The proliferation of different liver cancer cells was assessed by morphology and trypan blue assay. Cell apoptosis was confirmed by detecting phosphatidylserine (PS) externalization with Annexin V/PI double staining. The expression of related proteins was analyzed by Western bloting. Nude mouse model bearing subcutaneous transplanted BEL7402 tumor was established to study the therapeutic ability of Ad-pl4ARF. RESULTS: Ad-pl4ARF suppressed cell growth and proliferation, and promoted cell apoptosis of cancer cell lines with different genetic background. Ad-pl4ARF inhibited growth of liver cancer cells ( HepG2, BEL7402) in a dose-dependent manner. Ad-pl4ARF lead to overexpression of Bax and p21, the downstream regulating genes of p53. In the experimental therapy on nude mice bearing subcutaneous transplanted BEL7402 tumor, Ad-pl4ARF suppressed tumor growth significantly. CONCLUSION: pl4ARF is a short gene and with powerful function, which are consistent with the requirements for tumor suppressor genes used in gene therapy. It may play an important role in gene therapy against malignancies in the future.

• Administration of PUMA adenovirus increases the sensitivity of esophageal cancer cells to anticancer drugs.

  Authors: Haijuan Wang, Haili Qian, Jian Yu, Xueyan Zhang, Lin Zhang, Ming Fu, Xiao Liang, Qimin Zhan, Chen Lin

  Cancer biology & therapy. 05/2006; 5(4):380-5.

  Esophageal cancer is one of the most lethal human tumors, characterized by relative chemoresistance and poor prognosis. Researchers have been seeking for multimodality to improve its outcome ofEsophageal cancer is one of the most lethal human tumors, characterized by relative chemoresistance and poor prognosis. Researchers have been seeking for multimodality to improve its outcome of therapy. PUMA (p53 upregulated modulator of apoptosis) is a potent proapoptotic molecule that is rapidly induced in cells following DNA damage and is required for p53-induced apoptosis. We evaluated the therapeutic potential of PUMA adenovirus against esophageal cancer cell lines (KYSE-150, KYSE-410, KYSE-510 and YES-2). Infection with Ad-PUMA (PUMA Adenovirus) resulted in the more powerful cytotoxicity in these cell lines compared with Ad-p53. Furthermore, we assessed the efficacy of a combined treatment with Ad-PUMA and anticancer drug (cisplatin, paclitaxel, 5-fluorouracil, respectively) for these cells and found PUMA significantly increased the chemosensitivity of esophageal cancer cells, which may result from more abundant apoptosis induction. Interestingly, Ad-PUMA was found to be more efficient than Ad-p53 in inhibiting cell growth and enhancing the chemosensitivity of esophageal cancer cell lines irrespective of the p53 status. These results suggest that Ad-PUMA is a potent cytotoxic agent and could be a promising alternative in the cancer gene therapy in combination with chemotherapeutic agents.

• Enhancement of DNA vaccine potency by sandwiching antigen-coding gene between secondary lymphoid tissue chemokine (SLC) and IgG Fc fragment genes.

  Authors: Rongzhi Liu, Chunxia Zhou, Dongmei Wang, Wenbo Ma, Chen Lin, Yongquan Wang, Xiao Liang, Jie Li, Sujuan Guo, Yihua Wang, Youhui Zhang, Shuren Zhang

  Cancer biology & therapy. 05/2006; 5(4):427-34.

  DNA vaccine has become an attractive approach for generating antigen-specific immunity. Targeting antigens to FcRs for IgG (FcgammaRs) on dendritic cells (DCs) has been demonstrated to enhanceDNA vaccine has become an attractive approach for generating antigen-specific immunity. Targeting antigens to FcRs for IgG (FcgammaRs) on dendritic cells (DCs) has been demonstrated to enhance antigen presentation. Secondary lymphoid tissue chemokine (SLC) has been shown to increase immune responses not only by promoting coclustering of T cells and DCs in the lymph nodes and spleen but also by regulating their immunogenic potential for the induction of T cell responses. In this study, using HPV 16 E7 as a model antigen, we constructed a chemotactic-antigen plasmid DNA vaccine (pSLC-E7-Fc) by linking SLC and Fc gene sequences to each end of E7 and evaluated its potency of eliciting specific immune response. We found that immunization with pSLC-E7-Fc generated much stronger E7-specific lymphocyte proliferative and cytotoxic T lymphocyte (CTL) responses than control DNA. All the mice receiving pSLC-E7-Fc prophylactic vaccination remained tumor free upon subcutaneous inoculation of TC-1 cells, while those given control DNA all developed tumors. These tumor-free mice were also protected against TC-1 rechallenge. Complete tumor regression with long-term survival occurred in 72% of mice given pSLC-E7-Fc as therapeutic vaccination. In experimental lung metastasis model wherein TC-1 cells were intravenously injected, therapeutic vaccination with pSLC-E7-Fc significantly reduced the number of tumor nodules in the lung. In vivo depletion with antibodies against CD4+or CD8+ T cells both resulted in complete abrogation of the pSLC-E7-Fc-induced immunotherapeutic effect. Our data indicate that the DNA vaccine constructed by the fusion of SLC and IgG Fc fragment genes to antigen-coding gene is an effective approach to induce potent anti-tumor immune response via both CD4+ and CD8+ T cells dependent pathways.

• Enhancement of antitumour immunity by a novel chemotactic antigen DNA vaccine encoding chemokines and multiepitopes of prostate-tumour-associated antigens.

  Authors: Hanjun Qin, Chunxia Zhou, Dongmei Wang, Wenbo Ma, Xiao Liang, Chen Lin, Youhui Zhang, Shuren Zhang

  Immunology. 04/2006; 117(3):419-30.

  DNA vaccines provide an attractive technology against cancer because of their safety record in humans and ease of construction, testing and manufacture. In this study, several DNA fragments encodingDNA vaccines provide an attractive technology against cancer because of their safety record in humans and ease of construction, testing and manufacture. In this study, several DNA fragments encoding multiple cytotoxic T lymphocyte (CTL) and T helper cell epitopes were selected from human prostate-specific membrane antigen (hPSM), mouse prostatic acid phosphatase (mPAP), and human prostate-specific antigen (hPSA). These DNA fragments were ligated together to form a novel fusion gene, termed the 3P gene. The secondary lymphoid tissue chemokine (SLC), 3P and human immunoglobulin G Fc genes were inserted into pcDNA3.1 to construct a DNA vaccine, designated pSLC-3P-Fc. After vaccination, the DNA is taken up by cells that produce and secrete the SLC-3P-Fc fusion proteins, termed chemotactic antigen (chemo-antigen). The secreted chemo-antigens, in addition to promoting the co-localization of naive, non-polarized memory T cells and dendritic cells, are efficiently captured and processed by dendritic cells via receptor-mediated endocytosis and then cross-presented to both major histocompatibility complex class I and class II in a cognate manner. The results of this study demonstrate that vaccination with pSLC-3P-Fc by gene gun inoculation induced a strong antitumour response in a mouse tumour model, which significantly inhibited tumour growth and prolonged the survival time of the tumour-bearing mice. In vitro, the secreted SLC-3P-Fc fusion protein can attract lymphocytes from human peripheral blood mononuclear cells (PBMC); when human lymphocytes were stimulated by pSLC-3P-Fc-transfected autologous PBMC, CTLs were induced which could specifically kill hPSM-, hPAP-, or hPSA-expressing tumour cells. These observations provide a new vaccine strategy for cancer therapy through promoting the co-localization of lymphocytes and the concomitant enhancement of antigen-specific CD4+ helper and CD8+ cytotoxic T-cell responses against tumour.

• [The mechanism of resistance to 2', 2'-difluorodeoxycytidine (gemcitabine) in a pancreatic cancer cell line]

  Authors: Jie Yao, Feng-Yi Feng, Chen Lin, Xue-Yan Zhang, Ming Fu, Xiao Liang, Ying Yang

  Zhonghua zhong liu za zhi [Chinese journal of oncology]. 01/2006; 27(12):721-6.

  OBJECTIVE: To study the mechanism of resistance and its reversal to 2', 2'-difluorodeoxycytidine (gemcitabine) in a pancreatic cancer cell line SW1990. METHODS: Immunohistochemistry and RT-PCROBJECTIVE: To study the mechanism of resistance and its reversal to 2', 2'-difluorodeoxycytidine (gemcitabine) in a pancreatic cancer cell line SW1990. METHODS: Immunohistochemistry and RT-PCR techniques were used to investigate the mechanism of drug resistance. Salvicine (SAL) was used to reverse the drug resistance in a gemcitabine-resistant pancreatic cancer cell line (SW1990-GEM). RT-PCR, flow cytometry and MTT assay were employed to evaluate the effect of reversing drug resistance by salvicine. RESULTS: SW1990-GEM cells showed weak expression of P-glycoprotein (P-gp) revealed by immunohistochemistry, while its parental SW1990 cells were negative. Both cell lines did not express multidrug-resistance-related protein (MRP). As compared to the parental SW1990 cells, the mRNA expression of deoxycytidine kinase (dCK) was decreased while that of ribonucleotide reductase (RR) and mdr-1 was increased. With a concentration of 4 nmol/L, at one hr and 24 hr after SAL treatment, there was no change in mdr-1 mRNA expression, and the IC(50) of gemcitabine was 121.36 micromol/L and 121.64 micromol/L, respectively. However, when the concentration of SAL was increased to 30, 60, and 90 nmol/L, there was a dose-dependent down-regulation of mdr-1 mRNA expression in SW1990-GEM cells. The accumulation of rhodamine 123 was concomitantly increased, and the IC(50) of gemcitabine was correspondingly decreased. CONCLUSION: The resistance to gemcitabine of a pancreatic cancer cell line is due to decreased expression of dCK and increased expression of RR and mdr-1. Salvicine, only in toxic concentrations, can reverse the drug resistance by down-regulating mdr-1 gene and P-gp expression.