miR-466对HPV-16感染的宫颈癌细胞侵袭和迁移的影响

Influence of miR-466 on invasion and migration of cervical cancer cells induced by HPV-16

目的研究微小RNA-466(microRNA-466,miR-466)对人乳头瘤样病毒(Human papilloma virus,HPV)感染的宫颈癌细胞Siha增殖、迁移、侵袭的影响以及可能作用机制。方法 HPV-16^+ Siha细胞随机分为对照组、anti-NC组、anti-miR-466组;对照组细胞常规培养,anti-NC组和anti-miR-466组Siha细胞分别在Lipofectamine 2000诱导下转染阴性对照以及anti-miR-466质粒,荧光定量PCR(Quantitative polymerase chain reaction,qPCR)检测miR-466的表达量;细胞计数试剂盒(Cell counting kit-8,CCK-8)检测Siha细胞增殖,Transwell检测细胞的迁移、侵袭;在线软件TargetScan预测miR-466与信号转导和转录因子3(Signal transducer and activator of transcription 3,STAT3)的靶向关系,双荧光素酶报告基因进一步验证。结果与对照组(1.000±0.085)相比,anti-NC组(0.994±0.086)Siha细胞中miR-466表达量之间的差异无统计学意义;anti-miR-466组(0.263±0.040)Siha细胞中miR-466的表达量减少,差异具有统计学意义(P<0.05)。与对照组相比,anti-NC组Siha细胞的增殖、细胞迁移数和侵袭数之间的差异无统计学意义;anti-miR-466组Siha细胞的增殖、细胞迁移数(78.698±6.598)和侵袭数(43.658±4.658)减少,差异均具有统计学意义(P<0.05)。STAT3是miR-466下游靶基因。结论下调miR-466表达量可通过调控STAT3影响HPV-16^+ Siha细胞增殖、迁移、侵袭。

OBJECTIVE To explore the influence of microRNA-466(miR-466) on proliferation, migration and invasion of cervical cancer cell Siha induced by human papilloma virus(HPV) and observe its possible mechanisms of action. METHODS The HPV-16^+ Siha cells were randomly divided into the control group, the anti-NC group and the anti-miR-466 group. The control group was treated with routine culture, the Siha cells of the anti-NC group were induced by Lipofectamine 2000 and transfected into negative control, the Siha cells of the anti-miR-466 group were induced by Lipofectamine 2000 and transfected into anti-miR-466 plasmids, the expression of miR-466 was detected by using quantitative polymerase-chain-reaction(qPCR), the proliferation of the Siha cells was detected with the use of cell counting kit-8(CCK-8), and the migration and invasion of the cells were detected by means of Transwell. The targeting relationship between the miR-466 and the signal transducer and activator of transcription 3(STAT3) was predicted through online software TargetScan and was further validated by dual luciferase reporter gene. RESULTS There was no significant difference in the expression of miR-466 in Siha cells between the control group(1.000±0.085) and the anti-NC group(0.994±0.086);the expression of miR-466 in Siha cells of the anti-miR-466 group(0.263±0.040) was reduced, and there was significant difference( P<0.05). There were no significant differences in the proliferation, migrated or invaded counts of Siha cells between the anti-NC group and the control group;the proliferation, migrated(78.698±6.598) and invaded counts(43.658±4.658) of Siha cells of the anti-miR-466 group were reduced, and there were significant differences(P<0.05). STAT3 was a downstream target gene of miR-466. CONCLUSION Downregulation of the expression of miR-466 can affect the proliferation, migration and invasion of HPV 16^+ Siha cells by regulating STAT3.