长链非编码RNA ARAP1-AS1在肾透明细胞癌中的表达及作用研究

Expression and effect of long non-coding RNA ARAP1-AS1 in clear cell renal cell carcinoma

背景与目的:长链非编码RNA(long non-coding RNA,lncRNA)ARAP1-AS1在多种肿瘤中异常表达,但其在肾透明细胞癌(clear cell renal cell carcinoma,ccRCC)中的作用尚不清楚。探讨ARAP1-AS1在ccRCC中的生物学作用。方法:通过GEPIA数据库分析ARAP1-AS1在ccRCC组织中的表达及其与临床病理学特征及患者生存率的关系。采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR)检测ccRCC组织及邻近的非肿瘤组织中ARAP1-AS1的表达水平。将患者分为ARAP1-AS1高表达组和低表达组,分析ARAP1-AS1的表达水平与患者临床病理学特征之间的关系,并进行生存分析。通过细胞计数试剂盒-8(cell counting kit-8,CCK-8)实验、transwell迁移实验及侵袭实验检测ARAP1-AS1对ccRCC细胞体外增殖、迁移及侵袭能力的影响。采用蛋白质印迹法(Western blot)检测Wnt/β-catenin信号通路相关蛋白表达变化。采用BALB/c裸小鼠移植瘤模型分析ARAP1-AS1对ccRCC细胞体内成瘤能力的影响。结果:GEPIA数据库分析结果显示,ARAP1-AS1在ccRCC中高表达,且与患者肿瘤高分期及较差的生存率相关(P均<0.05)。RTFQ-PCR显示,ARAP1-AS1在ccRCC组织及细胞系中高表达,ARAP1-AS1的高表达与肿瘤大小和分期相关(P均<0.05)。ARAP1-AS1高表达患者的总生存率较差(P<0.05)。沉默ARAP1-AS1的表达可以抑制ccRCC细胞增殖、迁移和侵袭(P均<0.05)。沉默ARAP1-AS1可以降低Wnt/β-catenin信号通路相关蛋白的表达水平(P均<0.05)。沉默ARAP1-AS1可使ccRCC细胞体内成瘤能力减弱,并使Ki-67增殖指数降低。结论:ARAP1-AS1可通过激活Wnt/β-catenin信号通路促进ccRCC的进展。

Background and purpose:Long non-coding RNA(lncRNA)ARAP1-AS1 is abnormally expressed in a variety of tumors,but its role in clear cell renal cell carcinoma(ccRCC)is unknown.This study aimed to explore the biological role of ARAP1-AS1 in ccRCC.Methods:The expression level of ARAP1-AS1 in ccRCC tissues and its relationship to patient’s clinicopathological characteristics and survival rate were analyzed using the GEPIA database.The expression level of ARAP1-AS1 was measured in ccRCC and adjacent non-tumor tissues by real-time fluorescence quantitative polymerase chain reaction(RTFQPCR).Patients were divided into ARAP1-AS1 high and low expression groups,the relationship between ARAP1-AS1 expression level and patient’s clinicopathological characteristics was analyzed,and survival analysis was performed.The effect of ARAP1-AS1 in vitro proliferation,migration and invasion ability of ccRCC cells was determined by cell counting kit-8(CCK-8)assay,transwell migration and invasion assay.Changes in the expressions of Wnt/β-catenin signaling pathway-related proteins were detected by Western blot assay.The effect of ARAP1-AS1 on tumorigenic capacity in ccRCC cells in vivo was verified by tumor xenografts in nude mice.Results:The analysis of the GEPIA database showed that ARAP1-AS1 was highly expressed in ccRCC and associated with advanced tumor stage as well as poor survival in patients(all P<0.05).The results of RTFQ-PCR showed that ARAP1-AS1 was highly expressed in ccRCC tissues and cell lines,and high expression correlated with tumor size and stage(all P<0.05).The overall survival rate was poor in patients with high ARAP1-AS1 expression(P<0.05).Knockdown of ARAP1-AS1 expression inhibited the proliferation,migration and invasion of ccRCC cells(all P<0.05).Silencing of ARAP1-AS1 reduced the expression levels of the proteins involved in the Wnt/β-catenin signaling pathway(all P<0.05).Silencing of ARAP1-AS1 attenuated tumorigenic capacity in ccRCC cells and reduced Ki-67 proliferation index(P<0.05).Conclusion:ARAP1-AS1 promotes ccRCC progression through activation of the Wnt/β-catenin signaling pathway.