LINC00598靶向调控miR-381-3p影响宫颈癌细胞生物学行为的机制研究

Mechanism of LINC00598 on the Biological Behavior of Cervical Cancer Cells through Targeted Regulation of miR-381-3p Expression

目的探讨长链非编码RNA(lncRNA)LINC00598通过调控微小RNA-381-3p(miR-381-3p)表达影响宫颈癌细胞生物学行为的分子机制。方法采用qRT-PCR检测宫颈癌细胞系C33A、HeLa、SiHa、HCC94和人正常宫颈上皮细胞H8中LINC00598的表达。选取LINC00598表达最高的细胞系,分别转染无意义阴性对照序列(对照组)和LINC00598小分子干扰RNA(siRNA组)。qRT-PCR检测宫颈癌细胞中LINC00598表达。MTT法和Transwell侵袭实验分别检测宫颈癌细胞增殖情况和侵袭情况。DIANA数据库预测LINC00598的靶基因,双荧光素酶报告基因实验验证LINC00598与靶基因的调控关系。qRT-PCR和Western blot法检测LINC00598对靶基因表达的影响。结果宫颈癌细胞系中LINC00598表达显著高于人正常宫颈上皮细胞H8(P<0.05),LINC00598表达最高的细胞系是HeLa(P<0.01)。对照组和siRNA组LINC00598表达分别为1.05±0.19和0.27±0.04,差异有统计学意义(P<0.01)。与对照组比较,沉默LINC00598表达可显著抑制HeLa细胞增殖(P<0.05)和侵袭(P<0.01)。LINC00598可靶向结合miR-381-3p(P<0.01)。与对照组比较,沉默LINC00598表达可显著促进miR-381-3p的表达(P<0.01),抑制成纤维细胞生长因子7(fibroblast growth factor 7,FGF7)基因的表达(P<0.01)。结论LINC00598在宫颈癌细胞系中高表达,沉默LINC00598表达能够抑制宫颈癌HeLa细胞的增殖和侵袭,其作用机制可能是靶向上调miR-381-3p表达实现的。

Objective To explore the molecular mechanism of long non-coding RNA(lncRNA)LINC00598 that affects the biological behavior of cervical cancer cells by regulating the expression of microRNA-381-3p(miR-381-3p).Methods qRT-PCR was used to detect the expression of LINC00598 in cervical cancer cell lines C33A,HeLa,SiHa,HCC94 and human normal cervical epithelial cells H8.The cell line with the highest expression of LINC00598 was selected and transfected respectively with meaningless negative control sequences(control group)and LINC00598 small interfering RNA(siRNA group).qRT-PCR was used to detect the expression of LINC00598 in cervical cancer cells.The MTT method and Transwell invasion test were used to detect the proliferation and invasion of cervical cancer cells.The DIANA database predicted the target gene of LINC00598,and the dual luciferase reporter gene experiment verified the regulatory relationship between LINC00598 and the target gene.qRT-PCR and Western blotting were used to detect the effect of LINC00598 on target gene expression.Results The expression of LINC00598 in cervical cancer cell lines was significantly higher than that of normal human cervical epithelial cells H8(P<0.05).The cell line with the highest expression of LINC00598 was HeLa(P<0.01).The expression of LINC00598 in the control group and siRNA group was 1.05±0.19 and 0.27±0.04,respectively,and the difference was statistically significant(P<0.01).Compared with the control group,the silence of LINC00598 significantly inhibited Hela cell proliferation(P<0.05)and invasion(P<0.01).LINC00598 can target miR-381-3p(P<0.01).Compared with the control group,the silence of LINC00598 significantly promoted the expression of miR-381-3p(P<0.01)and inhibited the expression of fibroblast growth factor 7(FGF7)gene(P<0.01).Conclusion LINC00598 is highly expressed in cervical cancer cell lines.The silence of LINC00598 can inhibit the proliferation and invasion of cervical cancer HeLa cells.The mechanism may be achieved by targeting up-regulation of miR-381-3p expression.