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mTOR信号通路是细胞内重要的生命活动枢纽,它通过抑制细胞凋亡,促进细胞增殖来调控细胞生命活动。Rheb可以激活mTOR信号通路,进而参与多种肿瘤的发生发展。本研究以髓系白血病细胞株为研究对象,探讨Rheb在HL-60和K562中的作用及相关机制。本研究利用逆转录病毒技术使髓系白血病细胞株HL-60和K562过表达Rheb;利用Western blot和Real-Time PCR分别检测HL-60和K562细胞中Rheb的蛋白表达水平及mRNA表达水平;利用CCK-8法检测细胞活力;利用Annexin V-PE和7-AAD双染检测细胞凋亡。结果表明:成功构建了Rheb过表达的HL-60和K562细胞株,并发现Rheb过表达可以促进细胞生长;进一步研究发现,Rheb过表达加快细胞进入G2/M期(P<0.01),但不影响细胞凋亡。结论:Rheb通过加快细胞周期进程促进HL-60和K562细胞增殖。
The mTOR signaling pathway is an important intracellular life activity hub, which regulates cell life activities by inhibiting apoptosis and promoting cell proliferation. Rheb can activate the mTOR signaling pathway, and then participate in the development of many kinds of tumors. In this study, myeloid leukemia cell lines as the research object to explore the role of Rheb in HL-60 and K562 and related mechanisms. In this study, retroviral technique was used to overexpress Rheb in myeloid leukemia cell lines HL-60 and K562. The protein expression and mRNA expression of Rheb in HL-60 and K562 cells were detected by Western blot and Real-Time PCR respectively. Cell viability was determined by CCK-8 assay. Apoptosis was detected by Annexin V-PE and 7-AAD double staining. The results showed that the Rheb overexpression HL-60 and K562 cell lines were successfully constructed and Rheb overexpression was found to promote cell growth. Further studies showed that Rheb overexpression accelerated cells into G2 / M phase (P <0.01), but not Affect apoptosis. Conclusion: Rheb promotes the proliferation of HL-60 and K562 cells by accelerating cell cycle progression.