lncRNA HCG18调节miR-3619-5p/ITGB2轴对乳腺癌细胞迁移和侵袭的影响

Impacts of lncRNA HCG18 on migration and invasion of breast cancer cell by regulating miR-3619-5p/ITGB2 axis

目的探讨长链非编码RNA(lnc RNA)人类白细胞抗原复合体18(HCG18)能否通过调控微小RNA(mi R)-3619-5p/整合素β2(ITGB2)轴参与乳腺癌细胞的迁移和侵袭。方法体外培养人乳腺癌MCF-7细胞系及人乳腺上皮MCF-10A细胞系,qRT-PCR和Westernblot检测lncRNAHCG18、miR-3619-5p、ITGB2的表达水平;Starbase数据库预测lncRNA HCG18、ITGB2与miR-3619-5p的结合位点;利用转染技术将MCF-7细胞分为Control组、si-NC组、siHCG18组、si-HCG18+inhibitor NC组、si-HCG18+miR-3619-5p inhibitor组、miR-NC组、miR-3619-5p mimics组、miR-3619-5p mimics+pcDNA-NC组、miR-3619-5p mimics+ITGB2组,qRT-PCR和Western blot技术检测各组lncRNA HCG18、miR-3619-5p与ITGB2的表达水平,CCK-8、细胞划痕和Transwell实验分别检测细胞增殖活力、迁移能力和侵袭能力。结果与MCF-10A组比较,MCF-7组细胞lncRNA HCG18、ITGB2表达水平升高,miR-3619-5p表达水平下降(P<0.05);lncRNA HCG18、ITGB2与miR-3619-5p具有结合位点;沉默lncRNA HCG18表达或过表达miR-3619-5p可抑制MCF-7细胞增殖活力、迁移和侵袭,抑制miR-3619-5p表达或过表达ITGB2可部分逆转沉默lncRNAHCG18表达或过表达miR-3619-5p对MCF-7细胞增殖活力、迁移和侵袭的抑制作用(P<0.05)。结论lncRNAHCG18在乳腺癌细胞中表达上调,沉默lncRNA HCG18的表达可能通过上调miR-3619-5p表达、下调ITGB2表达抑制乳腺癌细胞迁移和侵袭。

Objective To investigate whether long non-coding RNA(lncRNA)human leukocyte antigen complex group 18(HCG18)can participate in the migration and invasion of breast cancer cells by regulating the microRNAs(miR)-3619-5p/integrinβ2(ITGB2)axis.Methods Human breast cancer cell line MCF-7 and human breast epithelial cell line MCF-10A were cultured in vitro.The expression levels of lncRNA HCG18,miR-3619-5p and ITGB2 were detected by qRT-PCR and Western blot.The Starbase database was applied to predict the binding sites of lncRNA HCG18,ITGB2,and miR-3619-5p.MCF-7 cells were grouped into Control group,si-NC group,si-HCG18 group,si-HCG18+inhibitor NC group,si-HCG18+miR-3619-5p inhibitor group,miR-NC group,miR-3619-5p mimics group,miR-3619-5p mimics+pcDNA-NC group,and miR-3619-5p mimics+ITGB2 group using transfection technology.qRT-PCR and Western blot techniques were applied to detect the expression levels of lncRNA HCG18,miR-3619-5p,and ITGB2,CCK-8,cell scratch and Transwell assay were used to detect cell proliferation,migration and invasion,respectively.Results Compared with the MCF-10A group,the expression levels of lncRNA HCG18 and ITGB2 in MCF-7 group was increased,while the expression level of miR-3619-5p was decreased(P<0.05).lncRNA HCG18,ITGB2 had binding sites with miR-3619-5p.Silencing lncRNA HCG18 expression or overexpression of miR-3619-5p was able to inhibit the proliferation,migration,and invasion of MCF-7 cells,inhibiting the expression of miR-3619-5p or overexpressing ITGB2 was able to partially reverse the inhibitory effects of silencing lncRNA HCG18 expression or overexpression of miR-3619-5p on the proliferation,migration,and invasion of MCF-7 cells(P<0.05).Conclusion The expression of lncRNA HCG18 is up-regulated in breast cancer cells,silencing the expression of lncRNA HCG18 may inhibit the migration and invasion of breast cancer cell migration by up-regulating the expression of miR-3619-5p and down-regulating the expression of ITGB2.