lncRNA PSMA3-AS1调节miR-224-3p/ARPP19轴对宫颈癌细胞恶性生物学行为的影响

Effect of lncRNA PSMA3-AS1 on the malignant biological behavior of cervical cancer cells by regulating the miR-224-3p/ARPP19 axis

目的探究长链非编码RNA人蛋白酶体α亚基3型反义RNA1(lncRNA PSMA3-AS1)调节微小RNA-224-3p(miR-224-3p)/环磷酸腺苷磷酸蛋白19(ARPP19)轴对宫颈癌细胞恶性生物学行为的影响。方法将宫颈癌HeLa细胞分为si-PSMA3-AS1组、si-NC组、si-PSMA3-AS1+miR-224-3p inhibitor组、si-PSMA3-AS1+inhibitor NC组和NC组。双荧光素酶报告基因实验验证lncRNA PSMA3-AS1、miR-224-3p、ARPP19的关系;qRT-PCR检测HeLa、人宫颈上皮细胞系HUCEC及宫颈癌组织lncRNA PSMA3-AS1、miR-224-3p表达;Western blot检测细胞、组织ARPP19蛋白水平;CCK8法检测HeLa细胞增殖;流式细胞术检测HeLa细胞凋亡;Transwell检测HeLa细胞侵袭、迁移;免疫细胞化学染色法检测γδT、Vδ1T阳性细胞数。结果lncRNAPSMA3-AS1、ARPP19在宫颈癌组织和HeLa细胞中高表达,miR-224-3p低表达;与NC组、si-NC组相比,si-PSMA3-AS1组HeLa细胞存活率、迁移细胞数量、侵袭细胞数量、lncRNA PSMA3-AS1水平、ARPP19水平、γδT阳性细胞数量、Vδ1T阳性细胞数量显著下降,HeLa细胞凋亡率显著升高(P<0.05),而抑制miR-224-3p表达减弱了沉默lncRNA PSMA3-AS1抑制HeLa细胞增殖、迁移、侵袭及促进凋亡的效果;lncRNA PSMA3-AS1负向调控miR-224-3p/ARPP19轴。结论lncRNA PSMA3-AS1的沉默抑制了HeLa细胞的恶性生物学进展,这种作用可能是通过调节miR-224-3p/ARPP19轴实现的。

Objective To explore the effect of long non-coding RNA human proteasomeαsubunit type 3 antisense RNA1(lncRNA PSMA3-AS1)on the malignant biological behavior of cervical cancer cells by regulating the microRNA-224-3p(miR-224-3p)/cyclic adenosine phosphate protein 19(ARPP19)axis.Methods Cervical cancer HeLa cells were divided into si-PSMA3-AS1 group,si-NC group,si-PSMA3-AS1+miR-224-3p inhibitor group,si-PSMA3-AS1+inhibitor NC group and NC group.The relationship between miR-224-3p and lncRNA PSMA3-AS1 or ARPP19 was verified by dual luciferase reporter gene assay.The expression of lncRNA PSMA3-AS1 and miR-224-3p in HeLa,human cervical epithelial cell line HUCEC,and cervical cancer tissue were detected by qRT-PCR.The level of ARPP19 protein in cells and tissues were detected by Western blot.CCK8 method was applied to detect HeLa cell proliferation;flow cytometry was applied to detect HeLa cell apoptosis;the invasion and migration of HeLa cell were detected by Transwell.Immunocytochemical staining was used to detect the numbers ofγδT and Vδ1T positive cells.Results In cervical cancer tissue and HeLa cells,lncRNA PSMA3-AS1 and ARPP19 were highly expressed,while miR-224-3p was expressed at a low level.compared with NC group and si-NC group,the survival rate of HeLa cells,numbers of migration and invasive cells,level of lncRNA PSMA3-AS1 and ARPP19,the numbers ofγδT and Vδ1T positive cells in si-PSMA3-AS1 group were obviously decreased,and apoptosis rate of HeLa cells was obviously increased(P<0.05).Inhibiting the expression of miR-224-3p weakened the effect of silencing lncRNA PSMA3-AS1 in inhibiting HeLa cell proliferation,migration,invasion,and promoting apoptosis.lncRNA PSMA3-AS1 negatively regulated the miR-224-3p/ARPP19 axis.Conclusion Suppression of lncRNA PSMA3-AS1 inhibits the malignant biological progression of HeLa cells,which may be achieved by regulating the miR-224-3p/ARPP19 axis.