微小RNA-324对非小细胞肺癌细胞迁移侵袭及ETS1表达的影响  

Effects of microRNA-324 on the migration and invasion ability and ETS1 expression of non-small cell lung cancer cells

作　　者：于法明[1] 姜东亮[1] 赵云[1] 刘传文[1] 杨国青[1] YU Faming;JIANG Dongliang;ZHAO Yun;LIU Chuanwen;YANG Guoqing(Department of Respiration, Puyang Oil field General Hospital, Puyang 457001, China)

机构地区：[1]濮阳市油田总医院呼吸科,河南濮阳457001

出　　处：《临床肿瘤学杂志》2018年第3期206-210,共5页Chinese Clinical Oncology

摘　　要：目的探讨微小RNA-324(miR-324)对非小细胞肺癌(NSCLC)细胞迁移和侵袭能力及E26转录因子1(ETS1)表达的影响。方法采用实时荧光定量PCR(QPCR)比较正常支气管上皮细胞16HBE和NSCLC细胞H322中miR-324水平的差异,采用脂质体Lipofectamine 2000法将miR-324抑制物或模拟物分别转染至H322细胞(抑制组和过表达组),以仅转染脂质体的细胞为空转染组;QPCR检测转染48 h后各组的miR-324水平,采用CCK-8法检测各组转染24、48、72 h的增殖情况,采用Transwell法检测各组的迁移和侵袭能力,Western blotting检测各组的ETS1表达情况。结果 QPCR检测结果显示H322细胞的miR-324水平为0.172±0.023,低于正常支气管上皮细胞16HBE的1.106±0.274,差异有统计学意义(P<0.05)。空转染组、抑制组和过表达组的miR-324水平分别为1.069±0.082、0.287±0.034和7.114±0.726,与空转染组相比,过表达组的miR-324水平升高而抑制组的miR-324水平降低,差异有统计学意义(P<0.05)。与空转染组相比,过表达组的增殖率降低而抑制组升高,差异有统计学意义(P<0.05)。Transwell检测结果显示迁移实验中空转染组、抑制组和过表达组的穿膜细胞数分别为(472.3±35.6)个、(692.1±50.6)个和(341.7±37.5)个,侵袭实验中空转染组、抑制组和过表达组的穿膜细胞数分别为(392.1±40.5)个、(521.6±57.6)个和(228.3±31.6)个,与空转染组相比,抑制组的穿膜细胞数升高而过表达组的穿膜细胞数降低,差异有统计学意义(P<0.05)。转染组、抑制组和过表达组的ETS1蛋白水平为0.56±0.13、0.87±0.23和0.26±0.09,与空转染组相比,抑制组的ETS1水平升高而过表达组的ETS1水平降低(P<0.05)。结论 miR-324在NSCLC细胞中的表达下调并可抑制细胞增殖和侵袭迁移能力,发挥类似抑癌基因的作用,可能通过靶向转录因子ETS1表达来发挥作用。Objective To explore the effect of microRNA-324（miR-324） on the migration and invasion ability and E26 transformation specific 1（ ETS1） expression of non-small cell lung cancer（ NSCLC） cells. Methods The real-time quantitative PCR（QPCR） was used to compare the level of miR-324 in normal bronchial epithelial cell 16 HBE and NSCLC cell H322. MiR-324 inhibitors or mimics were transfected into H322 cells（ inhibition group and overexpression group） by liposome Lipofectamine 2000,and cells only transfected with Lipofectamine 2000 were used as empty transfection group. QPCR was used to verify the miR-324 level in each group at 48 h after transfection. CCK-8 assay was used to detect the proliferative rates at 24,48 and 72 h after transfection. The migration and invasion abilities of each group were detected by Transwell method. The expression of ETS1 was detected by Western blotting at 48 h after transfection. Results QPCR detection showed that the level of miR-324 in H322 cells was 0. 172 ± 0. 023,lower than1. 106 ± 0. 274 in normal bronchial epithelial cells,and the difference was statistically significant（ P〈0. 05）. The levels of miR-324 were 1. 069 ± 0. 082,0. 287 ± 0. 034 and 7. 114 ± 0. 726 in empty transfection group,inhibition group and overexpression group,respectively. Compared with empty transfection group,the level of miR-324 increased in overexpression group but decreased in inhibition group（ P〈0. 05）. Compared with the empty transfection group,the proliferative rates decreased in overexpression group but increased in the inhibition group（ P〈0. 05）. Transwell assay showed that for empty transfection group,inhibition group and overexpression group,numbers of cell penetrating membrane were 472. 3 ± 35. 6,692. 1 ± 50. 6 and 341. 7 ± 37. 5 in the migration assay and 392. 1 ±40. 5,521. 6 ± 57. 6 and 228. 3 ± 31. 6 in the invasion assay. Compared with the empty transfection group,the number of penetrating membrane cells increased in the inhibition group but decrease

关 键 词：非小细胞肺癌 微小RNA-324 侵袭 迁移 E26转录因子1 

分 类 号：R734.2[医药卫生—肿瘤]