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目的:研究γ-分泌酶抑制剂DAPT对宫颈癌细胞系以及HPV16亚基因转化的永生化人宫颈上皮细胞系的作用,探讨DAPT临床治疗宫颈癌的可行性。方法:体外培养宫颈癌Siha、HeLa细胞系和HPV16亚基因转化的永生化人宫颈上皮细胞系(CRL2614)。加入不同浓度的γ-分泌酶抑制剂DAPT,不同时间点终止细胞生长,采用四甲基偶氮唑蓝(MTT)比色方法检测细胞生长情况;使用流式细胞计数仪检测细胞增殖周期中增殖指数(PI)、S期细胞比例(SPF)和凋亡细胞百分比。结果:应用MTT法检测细胞生长情况,5μmol/L及10μmol/LDAPT作用宫颈癌细胞系72h,可显著抑制其生长(P均<0.05)。10μmol/LDAPT对CRL2614细胞系作用72h无抑制作用(P>0.05)。流式细胞计数仪检测细胞增殖周期及细胞凋亡百分比。用5μmol/L及10μmol/LDAPT作用宫颈癌细胞系24,48,72h均可见SPF、PI明显降低,凋亡细胞百分比明显增加(P均<0.05)。10μmol/LDAPT作用CRL2614细胞系72h对SPF、PI及凋亡细胞百分比无影响(P>0.05)。结论:DAPT能抑制宫颈癌HeLa、Siha细胞系增殖,促进肿瘤细胞凋亡,对HPV16亚基因转化的永生化人宫颈上皮细胞系的增殖和凋亡没有明显影响。DAPT可能为药物治疗宫颈癌提供新思路。
Objective: To investigate the effect of γ-secretase inhibitor DAPT on cervical cancer cell line and immortalized human cervical epithelial cell line transformed by HPV16 subgenotype, and to investigate the feasibility of DAPT in the treatment of cervical cancer. Methods: The immortalized human cervical epithelial cell line (CRL2614) transformed with cervical cancer Siha, HeLa cell line and HPV16 subgenotype was cultured in vitro. Different concentrations of γ-secretase inhibitor DAPT were added, cell growth was stopped at different time points, cell growth was detected by MTT colorimetric method, and proliferation was detected by flow cytometry Index (PI), S phase cell fraction (SPF) and apoptotic cell percentage. Results: The cell growth was detected by MTT assay. The proliferation of cervical cancer cells was inhibited by 5μmol / L and 10μmol / L of LDAPT for 72h (all P <0.05). 10μmol / L LDAPT had no inhibitory effect on CRL2614 cell line for 72h (P> 0.05). Flow cytometry was used to detect cell cycle and percentage of apoptosis. The SPF and PI were obviously decreased and the percentage of apoptotic cells were significantly increased in both cervical carcinoma cell lines treated with 5μmol / L and 10μmol / L of LDAPT for 24, 48 and 72h (all P <0.05). The effect of 10μmol / L ODT on CRL2614 cell line for 72h had no effect on the percentage of SPF, PI and apoptotic cells (P> 0.05). CONCLUSION: DAPT can inhibit the proliferation of cervical cancer HeLa and SiH cell lines and promote the apoptosis of tumor cells, and has no obvious effect on the proliferation and apoptosis of immortalized human cervical epithelial cell line transformed by HPV16 subgenotype. DAPT may provide new ideas for drug treatment of cervical cancer.