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目的:探讨RNA干扰沉默人端粒酶反转录酶(hTERT)的表达后对卵巢癌SKOV3细胞增殖及化疗药物敏感性的影响。方法:化学合成hTERT siRNA及Control siRNA,用EntransterTM-R转染试剂将hTERT siRNA转染入卵巢癌SKOV3细胞。采用Real time-PCR法及Western blot法检测SKOV3细胞中hTERT mRNA及对应蛋白的表达;流式细胞术(FCS)检测细胞凋亡率;并分别用顺铂、阿霉素作用于SKOV3细胞,WST-1法检测细胞存活率及两种化疗药物的IC50。结果:与对照组相比,转染hTERT siRNA后,hTERT mRNA及蛋白表达量明显降低(P<0.05),细胞凋亡率明显增加,差异均有统计学意义(P<0.05),同时该组细胞的顺铂、阿霉素IC50显著下降。结论:hTERT siRNA不仅可以显著抑制卵巢癌SKOV3细胞中hTERT基因的表达,同时提高细胞对化疗药物的敏感性。
Objective: To investigate the effect of silencing human telomerase reverse transcriptase (hTERT) expression on the proliferation and chemosensitivity of ovarian cancer SKOV3 cells. METHODS: hTERT siRNA and Control siRNA were chemically synthesized and transfected into SKOV3 ovarian cancer cells using EntransterTM-R transfection reagent. The expression of hTERT mRNA and its corresponding protein in SKOV3 cells was detected by Real time-PCR and Western blot. The apoptosis rate was detected by flow cytometry (FCS). The cells were treated with cisplatin and doxorubicin respectively. -1 method to detect cell survival and IC50 of two chemotherapeutic drugs. Results: Compared with the control group, hTERT siRNA transfected hTERT siRNA significantly decreased the mRNA and protein expression of hTERT (P <0.05), and significantly increased the apoptosis rate (all P <0.05) Cell cisplatin, doxorubicin IC50 significantly decreased. Conclusion: hTERT siRNA can not only significantly inhibit the expression of hTERT gene in ovarian cancer SKOV3 cells, but also increase the sensitivity of cells to chemotherapeutic drugs.