Objective： The aim of our study was to observe the anti-tumor effect of silencing the expression of HIF-1α on cervical cancer in nude mice and to explore its mechanism of action. Methods： Human cervical cancer cell...Objective： The aim of our study was to observe the anti-tumor effect of silencing the expression of HIF-1α on cervical cancer in nude mice and to explore its mechanism of action. Methods： Human cervical cancer cell line Siha cells were divided into 3 groups： mock control group, control group transfected with scrambled sequence plasmid, and experimental group transfected with pU-HIF-la-shRNA eukaryotic expression plasmid. Cultured cells of the three groups were inoculated in nude mice to establish cervical cancer-bearing nude mice. HIF-la RNAi assay was performed to evaluate the tumor-suppressive effect of HIF-1α silencing on cervical cancer-bearing nude mice. Immunohistochemistry and Western blot were used to observe the distribution and protein expression of HIF-1α and GLUT1, while RT-PCR was adopted to detect the gene expression of HIF-1α, GLUTI and HK I1. The product of glycolysis （tactic acid） and apoptosis in tumor cells were detected by colorimetry and semi-quantitative TUNEL staining, respectively. Results： The tumor growth in experimental group was significantly slower than that in the two control groups （P 〈 0.05）. In the 50th day after transplantation, the tumor weight in the experimental group was （1.90 ± 0.28） g, significantly lower than （2.95 ± 0.77） g in the control group and （2.54 ± 0.56） g in the mock group （P 〈 0.01）. In the experimental group, the gene and protein levels of HIF-1α were 0.45 ± 0.04 and 1.25 ± 0.92, and the levels of GLUT1 were 0.32 ± 0.02 and 1.25 ± 0.48, respectively. Both indicators in HIF-la and GLUT1 were lower than that in the two control groups （P 〈 0.05）. The expression levels of HK Ⅱ gene and lactic acid in the experimental group were lower than that in the two control groups （P 〈 0.05）, but the apoptotic cells were much more numerous in the experimental group than that in matched control groups （P 〈 0.01）. Conclusion： The 9ene therapy by siRNAtargeted silencing of HIF-1α may down-regulate its downstream genes GLUT1 and HK Ⅱ expression, therefore, to reduce the tumor glycolysis activity and promote tumor cell apoptosis, and exert a tumor-suppressing effect in vivo.展开更多
Objective： The aim of our study was to investigate the effects of the anti-tumor composition of the acetoacetate extract of Vitex Negundo Seed （EVn-50） on the growth of human cervical carcinoma HeLa cells xenograft...Objective： The aim of our study was to investigate the effects of the anti-tumor composition of the acetoacetate extract of Vitex Negundo Seed （EVn-50） on the growth of human cervical carcinoma HeLa cells xenografts in nude mica and its possible molecular mechanism. Methods： Models of human cervical cancer HeLa cells xenografts transplanted subcuta- neously in nude mice were established and randomly divided into 7 groups （each group including 5 nude mice）： saline group, Taxol group, EVn-50 group, comp-6 group, comp-7 group, comp-8 group and comp-10 group. The volume and weight of Xe- nograts were observed and compared. The alteration of the weight of nude mice, and the change of serum levels ofLDH, ALT, Cr and WBC counts were examined and compared. The apoptotic rate of human cervical carcinoma HeLa cells xenografts was analyzed by FCM. The expressions of P53 and Bcl-2 proteins of HeLa cells xenografts were determined by Western blot- ting. Results： EVn-50 and its fractionated extracts could significantly suppress the increasing volume and weight of human cervical carcinoma HeLa cells xenografts in nude mice models in time-dependent manner, yet had no significant effect on the weight of nude mice, the serum levels of LDH, ALT, Cr and WBC were counted. When the xenografts were treated with EVn-50 and its fractionated extracts for 16 days, the apoptotic rate of xenografts cells were significantly increased, and the expression of P53 protein was up-regulated and protein level of Bcl-2 was decreased. Conclusion： EVn-50 and its fractionated extracts could suppress the growth of human cervical carcinoma HeLa cells xenografts in nude mice, which may be related to its pro- motion on xenografts cells apoptosis through down-regulation of Bcl-2 expressionPand activation of P53 expression.展开更多
文摘Objective： The aim of our study was to observe the anti-tumor effect of silencing the expression of HIF-1α on cervical cancer in nude mice and to explore its mechanism of action. Methods： Human cervical cancer cell line Siha cells were divided into 3 groups： mock control group, control group transfected with scrambled sequence plasmid, and experimental group transfected with pU-HIF-la-shRNA eukaryotic expression plasmid. Cultured cells of the three groups were inoculated in nude mice to establish cervical cancer-bearing nude mice. HIF-la RNAi assay was performed to evaluate the tumor-suppressive effect of HIF-1α silencing on cervical cancer-bearing nude mice. Immunohistochemistry and Western blot were used to observe the distribution and protein expression of HIF-1α and GLUT1, while RT-PCR was adopted to detect the gene expression of HIF-1α, GLUTI and HK I1. The product of glycolysis （tactic acid） and apoptosis in tumor cells were detected by colorimetry and semi-quantitative TUNEL staining, respectively. Results： The tumor growth in experimental group was significantly slower than that in the two control groups （P 〈 0.05）. In the 50th day after transplantation, the tumor weight in the experimental group was （1.90 ± 0.28） g, significantly lower than （2.95 ± 0.77） g in the control group and （2.54 ± 0.56） g in the mock group （P 〈 0.01）. In the experimental group, the gene and protein levels of HIF-1α were 0.45 ± 0.04 and 1.25 ± 0.92, and the levels of GLUT1 were 0.32 ± 0.02 and 1.25 ± 0.48, respectively. Both indicators in HIF-la and GLUT1 were lower than that in the two control groups （P 〈 0.05）. The expression levels of HK Ⅱ gene and lactic acid in the experimental group were lower than that in the two control groups （P 〈 0.05）, but the apoptotic cells were much more numerous in the experimental group than that in matched control groups （P 〈 0.01）. Conclusion： The 9ene therapy by siRNAtargeted silencing of HIF-1α may down-regulate its downstream genes GLUT1 and HK Ⅱ expression, therefore, to reduce the tumor glycolysis activity and promote tumor cell apoptosis, and exert a tumor-suppressing effect in vivo.
基金Supported by a grant from the Hengyang Municipal Science and Technology Programme(No.2011KJ36)
文摘Objective： The aim of our study was to investigate the effects of the anti-tumor composition of the acetoacetate extract of Vitex Negundo Seed （EVn-50） on the growth of human cervical carcinoma HeLa cells xenografts in nude mica and its possible molecular mechanism. Methods： Models of human cervical cancer HeLa cells xenografts transplanted subcuta- neously in nude mice were established and randomly divided into 7 groups （each group including 5 nude mice）： saline group, Taxol group, EVn-50 group, comp-6 group, comp-7 group, comp-8 group and comp-10 group. The volume and weight of Xe- nograts were observed and compared. The alteration of the weight of nude mice, and the change of serum levels ofLDH, ALT, Cr and WBC counts were examined and compared. The apoptotic rate of human cervical carcinoma HeLa cells xenografts was analyzed by FCM. The expressions of P53 and Bcl-2 proteins of HeLa cells xenografts were determined by Western blot- ting. Results： EVn-50 and its fractionated extracts could significantly suppress the increasing volume and weight of human cervical carcinoma HeLa cells xenografts in nude mice models in time-dependent manner, yet had no significant effect on the weight of nude mice, the serum levels of LDH, ALT, Cr and WBC were counted. When the xenografts were treated with EVn-50 and its fractionated extracts for 16 days, the apoptotic rate of xenografts cells were significantly increased, and the expression of P53 protein was up-regulated and protein level of Bcl-2 was decreased. Conclusion： EVn-50 and its fractionated extracts could suppress the growth of human cervical carcinoma HeLa cells xenografts in nude mice, which may be related to its pro- motion on xenografts cells apoptosis through down-regulation of Bcl-2 expressionPand activation of P53 expression.
