miRNA-16对人肺腺癌A549细胞增殖和VEGF-A表达水平的影响

发布时间：2018-07-03 11:29

本文选题：肺腺癌 + VEGF-A　；参考：《安徽医科大学》2016年硕士论文

【摘要】：目的探讨转染mi RNA-16后作用不同时间对体外培养的人肺腺癌A549细胞增殖的影响,以及对肺腺癌A549细胞内VEGF-A表达水平的影响。方法将研究对象(人肺腺癌A549细胞)分三组,分别是mi RNA-16转染组,阴性序列转染组,空白对照组。将50nmol/L的mi RNA-16 mimics和阴性对照序列用Lipofectamine 2000转染进人肺腺癌A549细胞中(空白对照组除了没有转染的RNA序列外,其他条件如所加的转染混合液,实验对象等都与其他两组一样),通过q RT-PCR检测转染后A549细胞中mi RNA-16的表达水平,通过四甲基偶氮唑蓝法(MTT法)检测转染mi RNA-16后不同时间点A549细胞的增殖抑制情况,通过ELSA法检测转染mi RNA-16并培养24小时后A549细胞上清液VEGF-A表达水平。结果q RT-PCR:转染培养24小时后,测得转染组mi RNA-16表达水平明显高于空白对照组和阴性对照组(P0.05),其中转染组表达水平是空白组的467.59±35.47倍,空白对照组和阴性对照组差异无统计学意义;MTT法:转染并培养后,转染组细胞增殖抑制率明显高于空白对照组和阴性对照组(P0.05),在24小时,mi RNA-16转染组、阴性序列转染组和空白对照组的OD值分别是0.471±0.028、0.561±0.020和0.567±0.025,在48小时,三组的OD值分别是0.543±0.021、0.678±0.026和0.688±0.022,而在72小时,三组的OD值分别是0.623±0.016、0.895±0.019和0.907±0.018;阴性对照组和空白对照组比,差异无统计学意义;ELISA法:转染并培养后,ELISA法测得转染组细胞上清VEGF-A表达水平为55.56±6.04 pg/ml,较阴性对照组(152.07±6.41 pg/ml)和空白对照组(146.43±7.71 pg/ml)明显降低(P0.05),而阴性对照组和空白对照组相比,差异无统计学意义。结论mi RNA-16能引起体外培养的人肺腺癌A549细胞增殖抑制,并且可能下调VEGF-A的表达。
[Abstract]:Objective to investigate the effects of transfection of mi RNA-16 on the proliferation of human lung adenocarcinoma A549 cells in vitro and the expression of VEGF-A in A549 cells. Methods Human lung adenocarcinoma A549 cells were divided into three groups: mi RNA-16 transfection group, negative sequence transfection group and blank control group. The 50 nmol / L mi RNA-16 mimics and the negative control sequence were transfected into human lung adenocarcinoma A549 cells by Lipofectamine 2000. The expression level of mi RNA-16 in A549 cells after transfection was detected by Q RT-PCR, and the proliferation inhibition of A549 cells was detected by MTT assay at different time points after transfection. The expression of VEGF-A in the supernatant of A549 cells was detected by ELSA method after transfection of mi RNA-16 and culture for 24 hours. Results Q RT-PCR: after 24 hours of transfection, the expression level of mi RNA-16 in the transfected group was significantly higher than that in the blank control group and the negative control group (P0.05), and the expression level in the transfected group was 467.59 卤35.47 times higher than that in the blank group. There was no significant difference between the blank control group and the negative control group in MTT assay: after transfection and culture, the cell proliferation inhibition rate of the transfected group was significantly higher than that of the blank control group and the negative control group (P0.05). The OD values of the negative sequence transfection group and the blank control group were 0.471 卤0.028, 0.561 卤0.020 and 0.567 卤0.025, respectively. At 48 hours, the OD values of the three groups were 0.543 卤0.021, 0.678 卤0.026 and 0.688 卤0.022, respectively, while at 72 hours, the OD values of the three groups were 0.623 卤0.0160.895 卤0.019 and 0.907 卤0.018, respectively. The expression of VEGF-A in the supernatant of the transfected group was 55.56 卤6.04 PG / ml, which was significantly lower than that of the negative control group (152.07 卤6.41 pg/ml) and the blank control group (146.43 卤7.71 pg/ml) (P0.05). The difference was not statistically significant. Conclusion mi RNA-16 can inhibit the proliferation of human lung adenocarcinoma cell line A549 in vitro and may down-regulate the expression of VEGF-A.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R734.2

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