Study on the role and mechanism of lncRNA NEAT1/miR-101-3p/RAC1 in the proliferation,migration and invasion of cervical cancer cells
Objective To investigate the mechanism of lncRNA NEAT1 in cervical cancer cells prolifera-tion,migration and invasion.Methods The expression of lncRNA NEAT1 in the cervical adenocarcinoma cell line HeLa,the cervical squamous carcinoma cell line SiHa and the human endometrial epithelial cell line EM was detected by RT-qPCR.Changes in proliferation,migration and invasion ability of cervical cancer cells were observed by CCK-8 and Transwell after interfering or overexpressing lncRNA NEAT1,and scanning electron microscopy to observe changes in cellular invadopodia.A dual luciferase reporter gene assay was used to detect the targeting relationship between lncRNA NEAT1 and miR-101-3p.The effects of lncRNA NEAT1/miR-101-3p/RAC1 on invasion,migration and proliferation of SiHa and HeLa cells were detected by immunofluorescence,Transwell,and CCK-8.Results Expression of lncRNA NEAT1 was significantly higher in cervical cancer HeLa(1.50±0.07,P＜0.05)and SiHa cells(7.59±0.10,P＜0.01)compared to control EM cells(0.99±0.04).Compared with the sh-NC group,cell prolifera-tion,migration,invasion and the number of invadopodias were significantly reduced in the sh-NEAT 1 group(with the addition of NEAT 1 knockdown plasmid)(P＜0.05).Compared with OE-NC group,the number of prolifera-tion,migration,invasion and invadopodias were significantly increased in SiHa and HeLa cells in OE-NEAT1 group(with the addition of NEAT1 overexpression plasmid)(P＜0.05).There was no significant change in lu-ciferase activity after co-transfection of miR-101-3p with mutant NEAT1(2.99±0.11,P＞0.05)compared to control(3.10±0.16),whereas co-transfection of miR-101-3p with wild-type NEAT1(1.15±0.05)resulted in a significant decrease in luciferase activity compared to control(2.97±0.13,P＜0.01),suggested that lncRNA NEAT1 binds to miR-101-3p target.miR-101-3p gene expression was significantly higher in the sh-NEAT 1 group(SiHa,3.86±0.39;HeLa,2.62±0.51,P＜0.01)compared to the sh-NC group(SiHa,0.97±0.09;HeLa,1.07±0.06),and significantly lower in the OE-NEAT1 group(SiHa,0.98±0.20;HeLa,1.05±0.18),miR-101-3p gene expression was significantly lower in OE-NEAT1 group(SiHa,0.35±0.05;HeLa,0.14±0.03,P＜0.05).Compared to OE-NEAT1 group,suggested that lncRNA NEAT1 is negatively correlated with miR-101-3p expression.The proliferation,migration and invasion of SiHa and HeLa cells were inhibited by siRNA-RAC1,miR-101-3p mimic(P＜0.05),and up-regulation of lncRNA NEAT1 partially reversed the above phenomena,resulting in an increase in the number of the cells proliferating,migrating and inva-ding(P＜0.05).Conclusion The expression of lncRNA NEAT1 in cervical cancer cells is significantly increased.As a competitive endogenous RNA(ceRNA)of miR-101-3p,lncRNA can competitively bind miR-101-3p and partially control its regulation of RAC1.Furthermore,it regulates the formation of inva-sive pseudopodia of cervical cancer cells and induces cell proliferation,migration and invasion.
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