论文部分内容阅读
目的:研究非小细胞肺癌(non-small cell lung cancer,NSCLC)A549细胞中miR-34a的表达情况,miR-34a和Met信号通路的相关性,进一步明确麦冬皂苷B(Ophiopogonin-B,OPB)治疗肺癌的分子机制。方法:采用非小细胞肺癌A549细胞,胚胎肺成纤维细胞MRC-5,以及A549+OPB细胞作为研究对象。实时荧光定量PCR(Quantitative Real-time PCR,qRT-PCR)方法检测miR-34a在A549,MRC-5,A549+OPB中的表达情况。MTT增殖实验检测miR-34a对A549细胞增殖能力的影响。使用生物信息学软件预测miR-34a的下游功能靶基因,通过荧光素酶报告验证。Western Blot技术验证miR-34a和OPB对Met蛋白表达的影响。结果:与MRC-5细胞相比,A549及A549+OPB细胞中miR-34a的表达量明显下降。转染miR-34a后可抑制A549细胞的增殖能力。生物信息学预测显示Met是miR-34a可能的功能性靶基因,且得到荧光素报告实验结果的证实。与未转染A549细胞株相,转染后的细胞株及A549+OPB细胞株,miR-34a表达升高,Met的表达降低。结论:在非小细胞肺癌中OPB可能通过miR-34a调控靶基因Met而发挥抗癌作用。
Objective: To investigate the expression of miR-34a and the relationship between miR-34a and Met signaling pathway in non-small cell lung cancer (NSCLC) A549 cells and further clarify the relationship between Ophiopogonin-B ) The molecular mechanism of treatment of lung cancer. Methods: Non-small cell lung cancer A549 cells, embryonic lung fibroblasts MRC-5, and A549 + OPB cells were used as research objects. The expression of miR-34a in A549, MRC-5, A549 + OPB was detected by quantitative real-time PCR (qRT-PCR) The effect of miR-34a on the proliferation of A549 cells was detected by MTT proliferation assay. Bioinformatics software was used to predict downstream functional target genes for miR-34a, validated by luciferase reporter assay. Western Blot technology to verify the impact of miR-34a and OPB on Met protein expression. Results: Compared with MRC-5 cells, the expression of miR-34a in A549 and A549 + OPB cells was significantly decreased. Transfection of miR-34a can inhibit the proliferation of A549 cells. Bioinformatics prediction shows that Met is a possible functional target gene of miR-34a and is confirmed by the results of fluorescein reporter assay. Compared with untransfected A549 cells, transfected cells and A549 + OPB cells, the expression of miR-34a increased and the expression of Met decreased. Conclusion: In non-small cell lung cancer, OPB may play an anti-cancer role by regulating the target gene Met with miR-34a.