LINC00662调节miR-424-5p/YAP1轴对卵巢癌细胞恶性生物学行为的影响

Effect of LINC00662 on the malignant biological behavior of ovarian cancer cells by regulating the miR-424-5p/YAP1 axis

目的探究长链非编码RNA LINC00662(LINC00662)调节微小RNA-424-5p(miR-424-5p)/Yes相关蛋白1(YAP1)轴对卵巢癌(OC)细胞恶性生物学行为的影响。方法用qRT-PCR法检测卵巢癌组织与细胞中LINC00662、miR-424-5p、YAP1 mRNA水平;双荧光素酶实验用来检测LINC00662与miR-424-5p及YAP1与miR-424-5p的靶向关系;将SKOV3细胞分为Control组、sh-NC、sh-LINC00662、sh-LINC00662+anti-NC、sh-LINC00662+anti-miR-424-5p,检测SKOV3细胞增殖、凋亡、迁移、侵袭能力;免疫印迹实验用来检测Ki67、caspase-3、YAP1及蛋白表达;裸鼠移植瘤用来检测LINC00662对卵巢移植瘤生长的影响。结果在卵巢组织及细胞中LINC00662、YAP1表达上调,miR-424-5p表达下调(P<0.05);通过生物信息学、双荧光素酶实验结果分析显示LINC00662与miR-424-5p及YAP1与miR-424-5p存在靶向结合位点;与sh-NC组相比,sh-LINC00662组细胞Edu阳性率、划痕愈合率、侵袭细胞数目、LINC00662、YAP1、Ki67表达水平降低,肿瘤体积和质量减小,细胞凋亡率、miR-424-5p、caspase-3表达水平升高(P<0.05);miR-424-5p低表达减弱了沉默LINC00662抑制SKOV3细胞恶性生物行为的作用。sh-LINC00662组肿瘤体积和质量比sh-NC组小,LINC00662、YAP1表达水平比sh-NC组低,miR-424-5p水平比sh-NC组高(P<0.05)。结论LINC00662在OC细胞和组织中显著上调,沉默LINC00662表达可能通过调节miR-424-5p/YAP1轴,抑制OC细胞增殖、迁移、侵袭,促进细胞凋亡。

Objective To investigate the effect of long non-coding RNA LINC00662(LINC00662)on the malignant biological behavior of ovarian cancer(OC)cells by regulating the microRNA-424-5p(miR-424-5p)/Yes associated protein 1(YAP1)axis.Methods qRT-PCR method was applied to detect the mRNA levels of LINC00662,miR-424-5p,and YAP1 in ovarian cancer tissues and cells.Dual luciferase assay was applied to detect the targeting relationship between LINC00662 and miR-424-5p,and between YAP1 and miR-424-5p.SKOV3 cells were separated into Control group,sh-NC,sh-LINC00662,sh-LINC00662+anti NC,sh-LINC00662+anti miR-424-5p.The proliferation,apoptosis,migration,invasion and protein expression of SKOV3 cells were detected.The effect of LINC00662 on the growth of ovarian xenografts was detected.Results LINC00662 and YAP1 expression were upregulated in ovarian tissue and cells,while miR-424-5p expression was downregulated(P<0.05).Bioinformatics and dual luciferase experiments reveled that LINC00662 had a targeted binding site with miR-424-5p,and YAP1 had a targeted binding site miR-424-5p.Compared with the sh-NC group,the positive rate of Edu,the rate of scratch healing,the number of invasive cells,the expression levels of LINC00662,YAP1 and Ki67 in the sh-LINC00662 group were decreased,the tumor volume and weight were decreased,and the apoptosis rate,the expression levels of miR-424-5p and caspase-3 were increased(P<0.05).Low expression of miR-424-5p attenuated the inhibitory effect of silencing LINC00662 on the malignant biological behavior of SKOV3 cells.Conclusion LINC00662 is obviously upregulated in OC cells and tissues.Silencing the expression of LINC00662 may inhibit OC cell proliferation,migration,invasion,and promote cell apoptosis by regulating the miR-424-5p/YAP1 axis.