miR-194对人非小细胞肺癌细胞系A549增殖、凋亡的影响及其机制  

Effects of miR-194 on proliferation and apoptosis of human non-small-cell lung cancer cell line A549 and its mechanism

作　　者：王纯斌[1,2] 袁琳 王梅娟[1,2] 沈福军 黎超 赵明宏 WANG Chunbin;YUAN Lin;WANG Meijuan;SHEN Fujun;LI Chao;Zhao Minghong(Yancheng Hospital Affiliated to Medical College of Southeast University, Yancheng 224001, China)

机构地区：[1]东南大学医学院附属盐城医院,江苏盐城224001 [2]盐城市第三人民医院 [3]南通大学附属建湖医院 [4]建湖县人民医院

出　　处：《山东医药》2019年第15期10-14,共5页Shandong Medical Journal

基　　金：江苏省自然科学基金资助项目(BK20161286)

摘　　要：目的 观察微小RNA-194(miR-194)对人非小细胞肺癌(NSCLC)细胞系A549增殖、凋亡的影响,并探讨其机制。方法 取A549细胞和人正常肺上皮细胞16HBE,采用qRT-PCR法检测两种细胞miR-194。常规培养A549细胞,经脂质体分别转染miR-194抑制物、miR-194模拟物(mimics)、对照48h(分别计为A、B、C组),采用qRT-PCR验证转染效果,MTS法检测细胞增殖能力,平板克隆实验检测细胞克隆形成能力,流式细胞仪检测细胞凋亡能力,Western blotting法检测细胞JAK2、pSTAT3、活化的Caspase-3蛋白。结果 A549、16HBE细胞中miR-194相对表达量分别为1.00±0.10、8.17±0.14,两者比较,P<0.05。A、B、C组miR-194相对表达量分别为0.31±0.11、8.35±0.31、1.00±0.10,A、B组分别与C组比较,P均<0.05。A组转染24、48、72h的OD值分别为0.494±0.069、0.843±0.084、1.111±0.109,B组分别为0.355±0.026、0.510±0.049、0.706±0.087,C组分别为0.427±0.022、0.664±0.068、0.906±0.060,A、B组分别与C组比较,P均<0.05。A、B、C组细胞克隆数目分别为(232.12±10.82)、(43.67±10.97)、(127.33±6.43)个,A、B组分别与C组比较,P均<0.05。与C组比较,A组细胞凋亡现象及数目明显减少,B组细胞凋亡现象及数目明显增多;A、B、C组细胞凋亡率分别为4.58%±1.01%、21.45%±1.78%、10.23%±1.65%,A、B组分别与C组比较,P均<0.05。A组JAK2、pSTAT3、活化的Caspase-3蛋白相对表达量分别为5.97±0.24、4.52±0.18、0.11±0.04,B组分别为0.34±0.03、0.21±0.02、7.21±0.79,C组分别为1.00±0.10、1.00±0.10、1.00±0.10,A、B组分别与C组比较,P均<0.05。结论 miR-194能抑制A549细胞的增殖、克隆形成能力,并诱导细胞凋亡,其机制可能与调控JAK2/STAT3信号通路并促进活化的Caspase-3蛋白表达有关。Objective To observe the effects of microRNA-194 (miR-194) on the proliferation and apoptosis of human non-small-cell lung cancer (NSCLC) cell line A549, and to explore its mechanism.Methods The miR-194 of A549 cells and human normal pulmonary epithelial cells 16HBE was detected by qRT-PCR. A549 cells were cultured routinely, and were transfecteded with miR-194 inhibitor, miR-194 mimics and controls by liposome for 48 h,(recorded as groups A, B and C, respectively). The transfection effect was verified by qRT-PCR. MTS assay was used to detect cell proliferation, plate cloning assay was used to detect the ability of cell cloning, the apoptotic ability was detected by flow cytometry, and JAK2, pSTAT3, and activated Caspase-3 protein was detected by Western blotting. Results The relative expression of miR-194 in A549 cells and 16HBE cells was 1.00±0.10 and 8.17±0.14, respectively, with statistically significant difference ( P <0.05). The relative expression of miR-194 in the groups A, B and C was 0.31±0.11, 8.35±0.31, and 1.00± 0.10, and statistically significant difference was found between groups A, B and group C ( P <0.05). OD at 24, 48 and 72 h after transfection was 0.494±0.069, 0.843±0.084, and 1.111±0.109, respectively, in the group A, 0.355 ± 0.026, 0.510±0.049, and 0.706±0.087, respectively, in the group B, and 0.427±0.022, 0.664±0.068, and 0.906 ±0.060, respectively, in the group C, with statistically significant difference (all P <0.05). The number of cell clones in the groups A, B and C were 232.12±10.82, 43.67±10.97, and 127.33±6.43, and statistically significant difference was found between groups A, B and group C ( P <0.05). Compared with group C, the apoptosis and number of cells in the group A was significantly reduced, while the apoptosis and number of cells in the group B significantly increased. The apoptotic rates of groups A, B and C were 4.58%±1.01%, 21.45%±1.78%, and 10.23%±1.65%, and statistically significant difference was found between groups A, B and group C ( P <0.05)

关 键 词：微小RNA-194 非小细胞肺癌 细胞增殖能力 细胞克隆形成能力 细胞凋亡能力 JAK2/STAT3信号通路 活化的Caspase-3蛋白 

分 类 号：R734.2[医药卫生—肿瘤]