论文部分内容阅读
该文采用阳离子脂质体Lipofectamine介导的方法将microRNA-34a转染入体外培养的人葡萄膜黑色素瘤细胞M23和SP6.5。应用BrdU法、细胞平板克隆形成实验检测转染microRNA-34a后对细胞增殖的影响,发现M23和SP6.5细胞增殖明显被抑制(P<0.01);并利用流式细胞技术检测转染microRNA-34a后细胞周期的变化,发现细胞停滞于G_1期;同时检测转染microRNA-34a后细胞caspase-3/7酶的活性,发现无明显改变。另外,Real-time PCR检测表明阿霉素处理后M23、SP6.5细胞中microRNA-34a的表达量上调(P<0.01)。用阿霉素处理转染microRNA-34a的M23、SP6.5细胞,检测caspase-3/7酶活性的改变,发现caspase-3/7酶活性显著增加(P<0.01)。本研究表明microRNA-34a通过抑制细胞周期来抑制体外培养的人葡萄膜黑色素瘤细胞的增殖,能够增加细胞对阿霉素的敏感性,但不直接诱导细胞凋亡。
In this paper, microRNA-34a was transfected into cultured human uveal melanoma cells M23 and SP6.5 by cationic liposome-mediated Lipofectamine. BrdU assay was used to detect the effect of microRNA-34a on the proliferation of M23 and SP6.5 cells. The proliferation of M23 and SP6.5 cells was significantly inhibited (P <0.01). Flow cytometry was used to detect the expression of microRNA- 34a after the cell cycle changes found in cells arrested in G_1 phase; at the same time detection of transfection of microRNA-34a caspase-3/7 enzyme activity and found no significant change. In addition, real-time PCR showed that the expression of microRNA-34a in M23 and SP6.5 cells was up-regulated after adriamycin treatment (P <0.01). The activity of caspase-3/7 was significantly increased in M23 and SP6.5 cells transfected with microRNA-34a with adriamycin (P <0.01). This study shows that microRNA-34a can inhibit the proliferation of human uveal melanoma cells in vitro by inhibiting the cell cycle, and can increase the sensitivity of cells to doxorubicin, but not directly induce apoptosis.