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目的:研究自噬基因Beclin 1对宫颈癌细胞株HeLa顺铂敏感性的影响并探讨可能的机制。方法:将自噬基因Beclin 1的真核表达载体pcDNA3.1(+)-Beclin1经脂质体包裹后转染人宫颈癌HeLa细胞。荧光定量PCR(RT-PCR)和Western blot分别检测Beclin 1及凋亡因子caspase-3 mRNA和蛋白的表达水平,MTT法检测顺铂对HeLa细胞的半数抑制浓度(IC50),Hoechest染色检测细胞凋亡。结果:pcDNA3.1(+)-Beclin1转染HeLa后,Beclin 1 mRNA和蛋白相对表达增加(P<0.05),caspase-3 mRNA和蛋白相对表达增加(P<0.05),顺铂对HeLa细胞的IC50由1.0μg/ml降至0.5μg/ml,HeLa细胞凋亡指数为(15.33±3.52)%,显著高于pcDNA3.1(+)组和未转染质粒组(P<0.05)。结论:自噬基因Beclin 1可以通过促进细胞凋亡提高宫颈癌HeLa细胞对顺铂的敏感性。
Objective: To investigate the effect of autophagy gene Beclin 1 on the sensitivity of cervical cancer cell line HeLa to cisplatin and to explore the possible mechanism. Methods: The eukaryotic expression vector pcDNA3.1 (+) - Beclin1 of autophagy gene Beclin 1 was encapsulated and transfected into HeLa cells. The mRNA and protein expressions of Beclin 1 and caspase-3 were detected by RT-PCR and Western blot respectively. The half inhibitory concentration (IC50) of cisplatin on HeLa cells was determined by MTT assay. Hoechst staining Death. Results: The mRNA and protein expressions of Beclin 1 increased (P <0.05) and the mRNA and protein expressions of Beclin 1 increased (P <0.05) after HeLa was transfected with pcDNA3.1 (+) - Beclin1. IC50 decreased from 1.0μg / ml to 0.5μg / ml. The apoptosis index of HeLa cells was (15.33 ± 3.52)%, which was significantly higher than that of pcDNA3.1 (+) group and untransfected plasmid group (P <0.05). Conclusion: The autophagy gene Beclin 1 can improve the sensitivity of cervical cancer HeLa cells to cisplatin by promoting apoptosis.