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目的探讨下调泛素结合酶E2-EPF对宫颈癌细胞株Caski细胞增殖、细胞周期及对放、化疗效果等方面的影响。方法采用RNA干扰技术下调Caski株E2-EPF的表达后,采用流式细胞术、MTT细胞增殖实验方法研究E2-EPF低表达对Caski株细胞生长速率的影响;通过不同剂量X线照射及常用化疗药物作用后,检测细胞凋亡及细胞增殖情况,了解下调E2-EPF对放、化疗的影响。结果通过瞬时转染E2-EPF-siRNA质粒载体,Caski株细胞内E2-EPF的mRNA水平下降至对照组的5%左右,其相应蛋白水平低至难以检出;与对照组相比,E2-EPF低表达Caski株细胞生长速率明显减低(P<0.05),对拓扑替康和依托泊苷药物的敏感性明显增强(P<0.05),但对放疗效果无明显影响(P>0.05)。结论 E2-EPF参与宫颈癌细胞株Caski的生长调控,瞬时下调E2-EPF可以抑制肿瘤细胞生长、增强细胞对拓扑异构酶I抑制剂和拓扑异构酶II抑制剂类化疗药物的敏感性。
Objective To investigate the effects of down-regulation of ubiquitin-conjugating enzyme E2-EPF on proliferation, cell cycle, radiotherapy and chemotherapeutic effect of cervical cancer cell line Caski. Methods The expression of E2-EPF in Caski strain was down-regulated by RNAi technique. The effects of low expression of E2-EPF on the cell growth rate of Caski strain were studied by flow cytometry and MTT cell proliferation assay. The effects of different doses of X-ray and common chemotherapy After the drug effect, the apoptosis and cell proliferation were detected to understand the influence of E2-EPF on radiotherapy and chemotherapy. Results The mRNA level of E2-EPF in Caski cells was decreased to about 5% of the control group by transient transfection of the E2-EPF-siRNA plasmid vector, and the corresponding protein level was low and difficult to detect. Compared with the control group, E2- (P <0.05). The sensitivity to topotecan and etoposide was significantly increased (P <0.05), but had no significant effect on radiotherapy (P> 0.05). Conclusions E2-EPF is involved in the growth and regulation of cervical cancer cell line Caski. Transient down-regulation of E2-EPF can inhibit the growth of tumor cells and enhance the sensitivity of cells to topoisomerase I and topoisomerase II inhibitors.