Aim To investigate the influences of melatonin (MT) on the growth of HeLa cells in vitro. Methods Theantiprolfferation activities of MT were evaluated in HeLa cells by means of trypan blue dye exclusion and MTT vital ...Aim To investigate the influences of melatonin (MT) on the growth of HeLa cells in vitro. Methods Theantiprolfferation activities of MT were evaluated in HeLa cells by means of trypan blue dye exclusion and MTT vital staining.The morphological changes of HeLa cells induced by MT were observed under transmission electronic microscope. Cell divisioncycle influenced by MT was assessed by a flow cytometry. Results MT produced a certain inhibition of HeLa cells at the con-centration of 2 mmol@ L-1 and prolonged the TD. The fraction of cells inhibited was 61.0%. The IC. so of HeLa cells exposed toMT for 96 h was 2.039 mmol@ L- 1. The flow cytometric analyses showed that exposure to MT for 72 h reduced the number ofHeLa cells in phase S. Under electronic microscope, the HeLa cells exposed to MT for 72 h displayed morphological changesof necrosis, apoptosis, more hetero-chromosome and less somatic chromosome. Conclusion MT showed certain influences onthe growth of HeLa cells. Its mechanism may probably be attributable to reduction of the number of cells in phase S.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
Objective： To study the radiosensitivity of SiHa cells with Ku80 inhibition by RNAi. Methods： pKu80-siRNA and pNeg-siRNA were constructed and then transfected into SiHa cells. Western blot and RT-PCR were applied to...Objective： To study the radiosensitivity of SiHa cells with Ku80 inhibition by RNAi. Methods： pKu80-siRNA and pNeg-siRNA were constructed and then transfected into SiHa cells. Western blot and RT-PCR were applied to measure the expression of Ku80. After 10 Gy irradiation with 6 MV X-ray, cells were harvested at 20 h, 28 h and 48 h, and analyzed by flow cytometry for apoptosis rate. The SF2 and α, β values of cells were acquired by clone formation array. Results： Two stable transfection cell clones SiHa/Ku80-siRNA and SiHa/Neg-siRNA were screened from the pKu80-siRNA and pNeg-siRNA transfected cells. Western blot and RT-PCR indicated that the expression of Ku80 was suppressed by the Ku80-siRNA, The apoptosis rates of SiHa/Ku80-siRNA were higher than control cells at 28 h and 48 h after X-ray irradiation （P 〈 0.05）. The SF2 value of SiHa/Ku80-siRNA （0.422） was lower than that of SiHa/Neg-siRNA cells （0.587） or untransfected cells （0.583）, and the a value of SiHa/Ku80-siRNA was 0.295 ± 0.016, higher than that of SiHa/Neg-siRNA （0.176 ± 0.010） or control cells （0.188 ± 0.011）. Conclusion： Inhibition of Ku80 could enhance the radiosensitivity of SiHa, which showed that Ku80 could be a good target for molecular therapy.展开更多
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
文摘Aim To investigate the influences of melatonin (MT) on the growth of HeLa cells in vitro. Methods Theantiprolfferation activities of MT were evaluated in HeLa cells by means of trypan blue dye exclusion and MTT vital staining.The morphological changes of HeLa cells induced by MT were observed under transmission electronic microscope. Cell divisioncycle influenced by MT was assessed by a flow cytometry. Results MT produced a certain inhibition of HeLa cells at the con-centration of 2 mmol@ L-1 and prolonged the TD. The fraction of cells inhibited was 61.0%. The IC. so of HeLa cells exposed toMT for 96 h was 2.039 mmol@ L- 1. The flow cytometric analyses showed that exposure to MT for 72 h reduced the number ofHeLa cells in phase S. Under electronic microscope, the HeLa cells exposed to MT for 72 h displayed morphological changesof necrosis, apoptosis, more hetero-chromosome and less somatic chromosome. Conclusion MT showed certain influences onthe growth of HeLa cells. Its mechanism may probably be attributable to reduction of the number of cells in phase S.
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.
文摘Objective： To study the radiosensitivity of SiHa cells with Ku80 inhibition by RNAi. Methods： pKu80-siRNA and pNeg-siRNA were constructed and then transfected into SiHa cells. Western blot and RT-PCR were applied to measure the expression of Ku80. After 10 Gy irradiation with 6 MV X-ray, cells were harvested at 20 h, 28 h and 48 h, and analyzed by flow cytometry for apoptosis rate. The SF2 and α, β values of cells were acquired by clone formation array. Results： Two stable transfection cell clones SiHa/Ku80-siRNA and SiHa/Neg-siRNA were screened from the pKu80-siRNA and pNeg-siRNA transfected cells. Western blot and RT-PCR indicated that the expression of Ku80 was suppressed by the Ku80-siRNA, The apoptosis rates of SiHa/Ku80-siRNA were higher than control cells at 28 h and 48 h after X-ray irradiation （P 〈 0.05）. The SF2 value of SiHa/Ku80-siRNA （0.422） was lower than that of SiHa/Neg-siRNA cells （0.587） or untransfected cells （0.583）, and the a value of SiHa/Ku80-siRNA was 0.295 ± 0.016, higher than that of SiHa/Neg-siRNA （0.176 ± 0.010） or control cells （0.188 ± 0.011）. Conclusion： Inhibition of Ku80 could enhance the radiosensitivity of SiHa, which showed that Ku80 could be a good target for molecular therapy.
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.
