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 this study was to study the inhibiting effect of survivin mRNA on transplanted XWLC-05 tumor on nude mice.Methods:We established XWLC-05 transplanted nude mice model.44 mice would be divided rando...Objective:The aim of this study was to study the inhibiting effect of survivin mRNA on transplanted XWLC-05 tumor on nude mice.Methods:We established XWLC-05 transplanted nude mice model.44 mice would be divided randomly into 4 groups:control group (blank),Lip group (simple liposome),survivin SODN group (transfected by sense oligonucleotide) and survivin ASODN group (transfected by antisense oligonucleotide).We would study general activities of nude mice in these 4 groups,measure the size of tumor and calculate the tumor inhabiting rate also.Pathological methods were applied in the analysis of the effect of different treatment on heart,kidney and liver of nude mice in these 4 groups.Results:Tumor grew slowly and size,weight of tumor was lower in survivin ASODN group when compared with that of others.Nude mice of survivin ASODN group showed lower growth index and tumor inhabiting rate was significantly higher than that of other groups (P < 0.05).Transplanted tumor on nude mice in control group (blank),Lip group,and survivin SODN group grew bigger as time passed and there was no significance among them (P > 0.05).We found a great deal of tumor cell necrosis in survivin ASODN group.No death of nude mice was observed in all 4 groups and we did not found obvious lesion in vital organs.Conclusion:Survivin ASDON could be used for the inhibitation of subcutaneously transplanted tumor in nude mice without obvious lesion in vital organs.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 grants from the Yunnan Provincial Scientific Office,Scientific Development Plan,Basic Research of Social Development (No.2009ZC120M)Key project of Social Development of Yunnan Provincial (No.2010CA015)United Fund of Science and Technology Bureau of Yunnan Province (No.2010CD185)
文摘Objective:The aim of this study was to study the inhibiting effect of survivin mRNA on transplanted XWLC-05 tumor on nude mice.Methods:We established XWLC-05 transplanted nude mice model.44 mice would be divided randomly into 4 groups:control group (blank),Lip group (simple liposome),survivin SODN group (transfected by sense oligonucleotide) and survivin ASODN group (transfected by antisense oligonucleotide).We would study general activities of nude mice in these 4 groups,measure the size of tumor and calculate the tumor inhabiting rate also.Pathological methods were applied in the analysis of the effect of different treatment on heart,kidney and liver of nude mice in these 4 groups.Results:Tumor grew slowly and size,weight of tumor was lower in survivin ASODN group when compared with that of others.Nude mice of survivin ASODN group showed lower growth index and tumor inhabiting rate was significantly higher than that of other groups (P < 0.05).Transplanted tumor on nude mice in control group (blank),Lip group,and survivin SODN group grew bigger as time passed and there was no significance among them (P > 0.05).We found a great deal of tumor cell necrosis in survivin ASODN group.No death of nude mice was observed in all 4 groups and we did not found obvious lesion in vital organs.Conclusion:Survivin ASDON could be used for the inhibitation of subcutaneously transplanted tumor in nude mice without obvious lesion in vital organs.
