miR-539通过抑素调控非小细胞肺癌的细胞增殖、迁移和凋亡

miR-539 for non-small cell lung cancer cell proliferation,migration and apoptosis by prohibitin regulation

目的:探讨miR-539通过抑素调控非小细胞肺癌(NSCLC)细胞增殖、迁移和凋亡的机制。方法:选择2019年1月至2020年1月期间武汉科技大学附属孝感医院收治的6例NSCLC手术患者作为研究对象进行回顾性分析,术中采集未实施放化疗的NSCLC标本,置于液氮中保存,提取总RNA,通过miR-539在NSCLC中靶基因的筛选确定抑素是miR-539的候选靶基因。采用反转录荧光定量聚合酶链式反应技术定量检测miR-539、抑素,CCK-8检测细胞增殖,划痕试验检测细胞迁移,流式细胞术检测细胞凋亡。比较靶向抑制抑素组、过表达miR-539组和低表达miR-539组的细胞增殖、迁移及凋亡情况检测。结果:过表达组抑素mRNA相对表达量(1.00±0.07)明显高于阴性对照组(0.20±0.02)(t=26.917,P<0.001),细胞增殖能力(4.21±1.21)较阴性对照组(8.09±0.97)降低(t=6.128,P<0.001),细胞迁移能力[(514.28±36.14)个]较阴性对照组[(1500.33±156.25)个]降低(t=15.060,P<0.001),细胞凋亡率[(53.21±7.59)%]较阴性对照组[(40.25±6.21)%]增高,(t=3.237,P=0.009)。抑制抑素组细胞增殖能力(11.32±0.36)较低表达组(9.03±0.28)和过表达组(5.69±0.08)高(F=672.994,P<0.001),细胞凋亡率[(38.56±5.24)%]较低表达组[(47.26±4.98)%]和过表达组[(58.61±6.32)%]低(F=19.735,P<0.001)。结论:抑素作为miR-539的候选靶基因,两者均可作为抑癌基因抑制癌细胞增殖、迁移,促进细胞凋亡。miR-539对NSCLC细胞增殖、迁移、凋亡的调控作用与抑素具有紧密的关系。

Objective To study miR-539 for non-small cell lung cancer(NSCLC)cell proliferation,migration and apoptosis by prohibitinregulation.Methods There were six cases of NSCLC operative patients from Xiaogan Hospital of Wuhan University of Science and Technology during the period January 2019 to January 2020 selected as subject,NSCLC samples not implement concurrent chemoradiation were collected intraoperatively and preserved in liquid nitrogen,total RNA were extracted,prohibit was candidate target genes of miR-539 while determined by miR-539 screen target genes in NSCLC.miR-539,prohibitin were used by RT-PCR technology quantitative detection,cell proliferation were detected by CCK-8,cell migration were detected by scratch test,cell apoptosis were detected by flow cytometry.The cell proliferation,migration and apoptosis among prohibitin inhibition group,over-express group,low-express group and negative control group were compared.Results The relative quantity of prohibitin in over-express group(1.00±0.07)was significantly higher than negative control group(0.20±0.02)(t=26.917,P<0.001),the cell proliferation(4.21±1.21)was significantly lower than negative control group(8.09±0.97)(t=6.128,P<0.001),the cell migration(514.28±36.14)was significantly lower than negative control group(1500.33±156.25)(t=15.060,P<0.001),the cell apoptosis rate[(53.21±7.59)%]was significantly higher than negative control group[(40.25±6.21)%](t=3.237,P=0.009).The cell proliferation of prohibitin inhibition group(11.32±0.36)was significantly higher than low-express group(9.03±0.28)and over-express group(5.69±0.08)(F=672.994,P<0.001),the cell apoptosis rate[(38.56±5.24)%]was significantly lower than low-express group[(47.26±4.98)%]and over-express group[(58.61±6.32)%](F=19.735,P<0.001).Conclusions Prohibitin can be as a candidate target genes of miR-539,both as a tumor suppressor gene transfer on promoting inhibit cancer cell proliferation,migration,promote apoptosis in NSCLC cell.