摘要
目的:探讨p53在肺癌PC-9细胞中对微小RNA-148b(mi R-148b)表达的影响、相应分子机制及对细胞增殖能力的影响。方法:以肺癌PC-9细胞作为研究对象,瞬时转染p53真核表达载体,建立高表达p53的实验模型。通过荧光定量PCR检测p53在PC-9细胞中对mi R-148b表达的影响。通过使用mi R-148b启动子荧光素酶报告基因表达载体,检测p53对mi R-148b启动子转录活性的影响。通过转染mi R-148b特异性抑制物(Inh-148b)低表达mi R-148b,使用CCK8法检测mi R-148b对p53在肺癌细胞增殖调控过程中的作用。结果:高表达p53可在肺癌PC-9细胞中促进mi R-148b的表达;高表达p53可增强mi R-148b启动子的活性;低表达mi R-148b可减弱p53对肺癌PC-9细胞增殖的抑制作用。结论:p53对肺癌细胞PC-9增殖的抑制作用部分依赖于mi R-148b。
Objective To explore the function of p53 on regulating the expression of miR-148b in lung cancer cell line PC-9 and its corresponding molecular mechanism and the impact on cell proliferation. Methods Transient transfection of p53 eukaryotic expressing plasmids into lung cancer cell line PC-9 was performed to establish a cell model over-expressing p53. RT-PCR was used to explicit the impact of p53 on the expression of miR-148b. A reporter vector containing miR-148b promoter was used to investigate the function of p53 on regulating the transcription of miR-148b. Low-expressing miR-148b by transfecting its specific inhibitors , a CCK-8 assay was performed to explore the influence of miR-148b on the lung cancer cell proliferation inhibited by p53. Results Over-expression of p53 promoted miR-148b expression in lung cancer cell line PC-9. P53 could increase the luciferase activity driven by miR-148b promoters. Knockdown of miR-148b attenuated the impact of p53 on inhibiting the proliferation of PC-9 cells. Conclusion P53 inhibits the proliferation of lung cancer cell line PC-9 partially depending on miR-148b.
出处
《实用医学杂志》
CAS
北大核心
2015年第12期1908-1911,共4页
The Journal of Practical Medicine
基金
国家973计划项目(编号:2009CB521802)
国家自然科学基金资助项目(编号:30973496)
国家青年科学基金项目(编号:81301899)