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目的:观察结肠癌转移相关基因1(MACC1)对膀胱癌细胞株T24增殖、侵袭能力以及对肝细胞生长因子信号通路(HGF/c-Met)的影响。方法:构建靶向过表达MACC1基因的慢病毒载体LV-MACC1-GFP,感染膀胱癌T24细胞株,经过筛选,建立稳定过表达MACC1基因的膀胱癌T24细胞株。通过实时荧光定量PCR、蛋白质印迹法(western blotting)分别检测MACC1、HGF、c-Met基因mRNA与蛋白表达量,噻唑蓝(MTT)比色法、Transwell体外细胞侵袭实验分别研究稳定过表达MACC1基因对细胞增殖和迁移的影响。结果:与阴性对照组相比,实时荧光定量PCR与western blotting证实,慢病毒LV-MACC1-GFP组MACC1、HGF、c-Met mRNA与蛋白表达量皆显著升高,Transwell细胞侵袭实验显示LV-MACC1-GFP组细胞迁移能力显著增强(P<0.05)。结论:慢病毒介导稳定过表达MACC1基因能使膀胱癌T24细胞株的增殖能力及侵袭能力增强,活化HGF/c-Met通路。
OBJECTIVE: To observe the effects of MACC1 on the proliferation and invasion of bladder cancer cell line T24 and on the hepatocyte growth factor signaling (HGF / c-Met). Methods: The lentiviral vector LV-MACC1-GFP targeting MACC1 gene was constructed and transfected into bladder cancer T24 cell line. After screening, we established bladder cancer T24 cell line stably overexpressing MACC1 gene. The mRNA and protein expression of MACC1, HGF and c-Met were detected by real-time fluorescence quantitative PCR and western blotting respectively. MTT assay and Transwell cell invasion assay were used to investigate the expression of MACC1 gene Effect on Cell Proliferation and Migration. Results: Compared with the negative control group, real-time quantitative PCR and western blotting confirmed that mRNA and protein expression of MACC1, HGF and c-Met in lentivirus LV-MACC1-GFP group were significantly increased. Transwell cell invasion assay showed that LV- The cell migration ability of MACC1-GFP group was significantly enhanced (P <0.05). Conclusion: The lentivirus-mediated overexpression of MACC1 gene can enhance the proliferation and invasion of bladder cancer T24 cell line and activate the HGF / c-Met pathway.