摘要
Abstract
Objective: To investigate the expression of UHRF1, the changes in cell proliferation and apoptosis after small interfering RNA (siRNA) for UHRF1 gene were transfected into the human ovarian cancer cell line OVCAR-3 , and to provide the basis for establishing UHRF1 gene as a new target for the treatment of ovarian cancer. Methods: Using the siRNA gene silencing technique, chemical synthesis small interfering RNA establishing for UHRF1 gene sequence were transfected into the human ovarian cancer cell line OVCAR-3 (transfection group). At the same time the negative control siRNA sequence was also transfected into the OVCAR-3 cell as the negative control group. Non-transfected cell was considered as the blank group. Real-time quantitative PCR and Western blot assay separately were used to compare the mRNA and protein level of UHRF1 before and after siRNA transfection. CCK-8 assay and flow cytometry were used to investigate the cell proliferation and cell apoptosis after UHRF1 gene silencing. Results: Real-time quantitative PCR results showed the expression of UHRF1 was significantly decreased in OVCAR-3 cells after the transfection of siRNA ( P<0. 01 ) . The Western blot results showed the relative expression between the UHRF1 protein and the (3-actin protein in transfection group, negative control group and blank group respectively were 0. 16+0. 04, 0. 33±0. 03 ,0. 35±0. 07. The UHRF1 protein level in transfection group was lower than negative control group and blank group,the difference was statistically significant (P<0.05). However, the comparison between the negative control group and the blank group was not statistically significant (P>0. 05). The experiment of CCK-8 and the flow cytometric analysis showed that UHRF1 gene silencing can effectively inhibit OVCAR-3 cell proliferation. 48 ,72,96 hours after transfection, the proliferation rates of transfected cells were lower than those of the negative control group and blank group, the difference was statistically significant (P<0, 01). At the same time, inhibition of UHRF1 gene expression can promote apoptosis. The apoptosis rate of transfected group,the negative control group were (15. 83±2. 22)% ,(7.46+2.93)% respectively, the difference was statistically significant (P<0. 01). Conclusion: Using the siRNA gene silencing techniques we can efficiently inhibit the expressions of UHRF1 in the OVCAR-3 cells, which results in decreased capacities of proliferation, and it can greatly promote their apoptosis. UHRF1 maybe become a new potential target for ovarian carcinoma treatment.关键词
UHRF1基因/卵巢肿瘤/OVCAR-3细胞株/RNA干扰/细胞增殖/凋亡Key words
UHRF1 gene/Ovarian neoplasms/OVCAR-3 cell line/RNA interference/Cell proliferation/ Apoptosis分类
医药卫生
