circDOCK4促进肾透明细胞癌细胞增殖、迁移和侵袭的研究

circDOCK4 promotes the proliferation,migration and invasion of clear cell renal cell carcinoma

目的探索在肾透明细胞癌(ccRCC)中高表达的circDOCK4对ccRCC细胞增殖、迁移和侵袭的影响。方法实时荧光定量PCR检测circDOCK4在ccRCC组织和细胞系ACHN、Caki-1的表达水平;核酸电泳和Rnase R消化实验验证circDOCK4的成环性;EdU、MTS和克隆形成实验检测circDOCK4对ACHN、Caki-1的增殖能力的影响;Transwell小室实验和划痕愈合实验检测circDOCK4对ACHN、Caki-1的迁移和侵袭能力的影响;核浆分离实验检测circDOCK4在ACHN、Caki-1的细胞亚定位。结果circDOCK4在ccRCC组织和细胞系ACHN、Caki-1中显著高表达。核酸电泳和Rnase R消化实验证实circDOCK4的成环性。沉默circDOCK4的表达可明显抑制ACHN和Caki-1的增殖、迁移和侵袭能力。核浆分离实验提示circDOCK4主要定位于细胞浆。结论circDOCK4具有促进ccRCC细胞增殖、迁移和侵袭的能力,有望成为ccRCC的生物标志物或治疗靶点。

Objective To investigate the proliferation,migration and invasion function of the highly-expressed circDOCK4 in clear cell renal cell carcinoma(ccRCC).Methods The expression of circ-DOCK4 in ccRCC tissues and cell lines(ACHN,Caki-1)were detected by quantitative real time PCR(qRT-PCR).Nucleic acid electrophoresis and Rnase R treatment were employed to verify the cyclonicity of circDOCK4.The EdU assay,MTS assay and Colony formation assay were applied to validate the roles of circDOCK4 in proliferation in ACHN and Caki-1.Transwell assay and Wound-healing assay were per-formed to evaluate the effects of circDOCK4 on cellular migration and invasion ability in ACHN and Caki-1.Nuclear mass separation assay was used to identify the subcellular location of circDOCK4 in ACHN and Caki-1.Results The research results demonstrated that circDOCK4 was highly expressed both in ccRCC tissues and cell lines.Nucleic acid electrophoresis and Rnase R treatment verified the cyclo-nicity of circDOCK4.Downregulating the expression of circDOCK4 could suppress the proliferation,migra-tion and invasion ability in ACHN and Caki-1.Nuclear mass separation assay confirmed that circDOCK4 was predominantly located in cytoplasm.Conclusion circDOCK4 could promote the proliferation,migra-tion and invasion ability in ccRCC,which may serve as a biomarker or target for ccRCC.