长链非编码RNA CCAT1靶向调控miR-377对前列腺癌PC3细胞生物学行为的影响

Effects of long non-coding RNA CCAT1 on the biological behavior of prostate cancer PC3 cells by targeting miR-377

目的探讨长链非编码RNA CCAT1靶向调控miR-377对前列腺癌PC3细胞生物学行为的影响。方法选取前列腺癌组织78例为病理组,同时获取78例相邻的非恶性组织标本为癌旁组织,应用RT-PCR法测定2组标本miR-377、InRNA CCAT1表达水平,分析不同临床病理特征前列腺癌患者IncRNA RNA CCAT1、miR-377表达水平差异。设PC3前列腺癌细胞组、InRNA-NC组、InRNA CCAT1 mimics组、InRNA CCAT1 inhibitor组。培养72 h后,采用CCK-8测定法检测细胞增殖水平,流式细胞仪检测细胞凋亡水平,transwell测定法评估细胞侵袭能力,RT-PCR法及蛋白印迹法测定细胞miR-377、InRNA CCAT1、RUNX1表达水平。结果病理组miR-377水平低于癌旁组,InRNA CCAT1水平高于癌旁组(P<0.05)。病理学分级越高、TNM分期越高、有淋巴血管间隙浸润、有淋巴结转移、有复发的前列腺癌患者miR-377表达水平越低,InRNA CCAT1表达水平越高(P<0.05)。与PC3前列腺癌细胞组、InRNA-NC组比较,InRNA CCAT1 mimics组细胞OD值、存活率、穿膜数、InRNA CCAT1、凋亡率升高,miR-377、RUNX1蛋白降低(P<0.05),InRNA CCAT1 inhibitor组细胞OD值、存活率、穿膜数、InRNA CCAT1、凋亡率降低,miR-3771、RUNX1蛋白升高(P<0.05);与InRNA CCAT1 mimics组比较,InRNA CCAT1 inhibitor组细胞OD值、存活率、穿膜数、InRNA CCAT1、凋亡率降低,miR-377、RUNX1蛋白升高(P<0.05)。结论InRNA CCAT1在前列腺癌组织中高表达,miR-377在前列腺癌组织中低表达,InRNA CCAT1、miR-377表达水平与前列腺癌患者临床病理特征具有一定相关性。InRNA CCAT1靶向抑制miR-377、RUNX1表达进而促进前列腺癌PC3细胞恶性生物学行为。

Objective To explore the effect of long non-coding RNA(lncRNA)CCAT1 on the biological behavior of prostate cancer PC3 cells by targeting microRNA-377(miR-377).Methods Totally 78 cases of prostate cancer tissue in our hospital were selected as the pathological group,while 78 adjacent non-malignant tissues were obtained as the adjacent group.The expression levels of miR-377 and lncRNA CCAT1 in the two groups were measured by reverse transcription-polymerase chain reaction(RT-PCR).The differences in the expression levels of lncRNA CCAT1 and miR-377 in prostate cancer patients with different clinical and pathological characteristics were analyzed.Four experimental groups were established,including PC3 prostate cancer cell group,lncRNA NC group,lncRNA CCAT1 mimics group,and lncRNA CCAT1 inhibitor group.After 72 hours of culture,cell proliferation was detected by CCK-8 assay,cell apoptosis was detected by flow cytometry,and cell invasion was measured by transwell assay.Meanwhile,the mRNA and protein expression levels of miR-377,lncRNA CCAT1 and RUNX1 were measured using RT-PCR and Western blotting,respectively.Results The level of miR-377 in the pathological group was lower than that in the adjacent group,while the level of lncRNA CCAT1 was higher than the adjacent group(P<0.05).The expression level of miR-377 in prostate cancer patients with higher pathological grade,higher TNM stage,lymphovascular space infiltration,lymph node metastasis and recurrence was significantly lower,whereas the expression level of lncRNA CCAT1 was significantly higher(P<0.05).Compared with PC3 prostate cancer cell group and lncRNA NC group,OD value,survival rate,the number of penetrating membranes,lncRNA CCAT1 expression,and apoptosis rate in lncRNA CCAT1 mimics group increased obviously,while the protein expressions of miR377 and RUNX1 decreased remarkably(P<0.05).The lncRNA CCAT1 inhibitor group showed significantly decreased OD value,survival rate,number of penetrating membranes,lncRNA CCAT1 expression and apoptosis rate,as well as up-regulated protein expressions of miR-3771 and RUNX1(P<0.05).Compared with lncRNA CCAT1 mimics group,OD value,survival rate,number of penetrating membranes,lncRNA CCAT1 expression and apoptosis rate in the lncRNA CCAT1 inhibitor group were significantly lower,and miR-377 and RUNX1 protein expressions were significantly higher(P<0.05).Conclusion LncRNA CCAT1 is highly expressed in prostate cancer tissues,while miR-377 is lowly expressed in prostate cancer tissues.The expression levels of lncRNA CCAT1 and miR-377 are correlated with clinicopathological characteristics of prostate cancer patients.Targeted inhibition of miR-377 and RUNX1 expression by lncRNA CCAT1 promotes the malignant biological behaviors of prostate cancer PC3 cells.