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
To investigate the effect of Yikun Neiyi Wan (益坤内异丸YKNYW) and gestrinone on the expression of aromatase P450 (P450arom), cyclo-oxygenase-2 (COX-2) and estrogen receptor (ER) in isolated ectopic and normal...To investigate the effect of Yikun Neiyi Wan (益坤内异丸YKNYW) and gestrinone on the expression of aromatase P450 (P450arom), cyclo-oxygenase-2 (COX-2) and estrogen receptor (ER) in isolated ectopic and normal endometriat stroma cells in vitro. Methods: Digestion and serial filtration were used to isolate and culture the ectopic and eutopic endometrial cells from patients with chocolate cyst in virto Transformation of the cell morphology was observed in a inverted microscope. The effect of YKNYW on the expression of aromatase P450, cyclo-oxygenase-2, estrogen receptor in cultured endometriosis cells were detected by immunohistochemical method. Results: The expression levels of P450arom, COX-2 in glandular epithelium cells in vitro were decreased significantly by YKNYW compared with gestrinone (P〈0.05). ER expression in mesenchymal cells of endometriosis was increased by YKNYW in the large and medium dosage groups compared with gestrinone. Conclusion: The mechanism by which YKNYW alleviates endometriosis pain is possibly related to the decrease in ectopic endometrial P450 arom and COX-2 expression in glandular epithelium, contrary to gestrinone, and the increase in ER expression in mesenchymalis, consistent with gestrione in patients with endometriosis.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.
文摘To investigate the effect of Yikun Neiyi Wan (益坤内异丸YKNYW) and gestrinone on the expression of aromatase P450 (P450arom), cyclo-oxygenase-2 (COX-2) and estrogen receptor (ER) in isolated ectopic and normal endometriat stroma cells in vitro. Methods: Digestion and serial filtration were used to isolate and culture the ectopic and eutopic endometrial cells from patients with chocolate cyst in virto Transformation of the cell morphology was observed in a inverted microscope. The effect of YKNYW on the expression of aromatase P450, cyclo-oxygenase-2, estrogen receptor in cultured endometriosis cells were detected by immunohistochemical method. Results: The expression levels of P450arom, COX-2 in glandular epithelium cells in vitro were decreased significantly by YKNYW compared with gestrinone (P〈0.05). ER expression in mesenchymal cells of endometriosis was increased by YKNYW in the large and medium dosage groups compared with gestrinone. Conclusion: The mechanism by which YKNYW alleviates endometriosis pain is possibly related to the decrease in ectopic endometrial P450 arom and COX-2 expression in glandular epithelium, contrary to gestrinone, and the increase in ER expression in mesenchymalis, consistent with gestrione in patients with endometriosis.