长链非编码RNA LINC00662通过调控miR-409-5p对宫颈癌细胞增殖和侵袭的影响

Effects of long-chain non-coding RNA LINC00662 on proliferation and invasion of cervical cancer cells by regulating the expression of miR-409-5p

目的探讨长链非编码RNA LINC00662对miR-409-5p表达的调控作用及对宫颈癌细胞增殖和侵袭的影响。方法荧光实时定量聚合酶链式反应(qPCR)检测宫颈癌组织、宫颈癌细胞系中LINC00662的表达。转染载有LINC00662 的质粒或阴性对照质粒至LINC00662 表达最低的细胞系。细胞计数(CCK-8)法和Transwell侵袭实验分别检测高表达LINC00662对细胞侵袭能力的影响。采用生物信息学技术预测LINC00662 的下游基因,qPCR检测miR-409-5p和DIRAS3 mRNA的表达量,Western Blot检测DIRAS3蛋白的表达量。结果LINC00662在宫颈癌组织和宫颈癌细胞系中表达明显减少(P<0.05)。高表达LINC00662 明显抑制宫颈癌HCC94细胞的增殖能力(P<0.05)和侵袭能力(P<0.01)。生物信息学技术显示LINC00662可靶向结合miR-409-5p,miR-409-5p可靶向结合DIRAS 家族GTP酶3(DIRAS familyGTPase 3,DIRAS3)基因。高表达LINC00662后,miR-409-5p表达减少(P<0.01),DIRAS3在mRNA和蛋白水平的表达均增加(P<0.01)。结论LINC00662在宫颈癌组织和细胞系呈低表达,高表达LINC00662可通过影响miR-409-5p的表达抑制宫颈癌HCC94细胞的增殖和侵袭,在宫颈癌中表现为抑癌基因的功能。

Objective To investigate the regulation of long-chain non-coding RNA LINC00662 on the expression of miR-409-5p and its effects on proliferation and invasion of cervical cancer cells.Methods Fluorescence real-time quantitative polymerase chain reaction(qPCR)was used to detect the expression levels of LINC00662 in 39 cervical cancer tissues and adjacent tissues,cervical cancer cell lines,and normal cervical epithelial cell line. Plasmid carrying the LINC00662 or negative control plasmid was transfected into the cell line with the lowest expression of LINC00662 and the transfection efficiency was detected by qPCR. Cell count(CCK-8)method was used to detect the effect of high expression of LINC00662 on the proliferation of cervical cancer cells. Transwell invasion assay was used to detect the effect of high expression of LINC00662 on cell invasion. The downstream gene of LINC00662 was predicted by bioinformatics technology. The expression of miR-409-5p and DIRAS3 mRNA was detected by qPCR,and the expression of DIRAS3 protein and related protein was detected by Western Blot.Results The expression of LINC00662 was significantly decreased in cervical cancer tissues and cervical cancer cell lines(P < 0.01). High expression of LINC00662 significantly inhibited the proliferation ability of cervical cancer HCC94 cells(P < 0.01)as well as invasion ability(P < 0.01). Bioinformatics technology showed that LINC00662 can target miR-409-5p,and miR-409-5p can target the DIRAS family GTPase 3(DIRAS3)gene. After high expression of LINC00662,the expression of miR-409-5p was decreased(P < 0.01). The expression of DIRAS3 was significantly increased at both mRNA and protein levels.(P < 0.01).Conclusions Cervical cancer tissues and cell lines have low expression of LINC00662. High expression of LINC00662 can inhibit the proliferation and invasion of cervical cancer HCC94 cells by affecting the expression of miR-409-5p,and therefore may function as a tumor suppressor gene.