circ_0091579/miR-330-3p/RNF126轴影响结直肠癌LoVo细胞的增殖、侵袭和凋亡

circ_0091579/miR-330-3p/RNF126 axis affects the proliferation,invasion and apoptosis of colorectal cancer LoVo cells

目的:探讨环状RNA circ0091579作为分子海绵吸附miR-330-3p介导环指蛋白126(ring finger protein 126,RNF126)对结直肠癌(colorectal cancer,CRC)细胞增殖、凋亡、侵袭的影响。方法:选取2019年2月至2020年5月在大理大学第一附属医院行手术治疗的60例CRC患者的癌组织和癌旁组织。构建circ0091579、miR-330-3p的过表达或敲减的CRC LoVo细胞,qPCR检测CRC组织和细胞中circ0091579、miR-330-3p和RNF126的表达;MTT、Transwell、流式细胞术分别检测细胞的增殖、侵袭、凋亡情况;生物信息学方法预测circ0091579和miR-330-3p、miR-330-3p和RNF126的靶向关系并用双荧光素酶报告实验和RNA免疫沉淀实验验证。结果:CRC组织和多种细胞(HCT116、SW620、CW-2、LoVo细胞)中,circ0091579和RNF126均高表达、miR-330-3p均低表达(均P<0.05)。敲减circ0091579可以抑制LoVo细胞的增殖、侵袭而促进其凋亡(均P<0.05),但该影响在加入miR-330-3p后被逆转;过表达miR-330-3p使LoVo细胞增殖和侵袭能力减弱但凋亡程度加强(均P<0.05),该影响在加入RNF126后被抵消。circ0091579、miR-330-3p和RNF126之间存在靶向作用关系。结论:circ0091579通过mi R-330-3p/RNF126轴促进LoVo细胞增殖、迁移和侵袭而抑制其凋亡。

Objective:This study is designed to identify the effect of circRNA circ0091579 on the proliferation,apoptosis,invasion of colorectal cancer(CRC)cells by functioning as a molecular sponge of miR-330-3 p and mediating RNF126.Methods:Sixty pairs of cancer tissue and adjacent normal tissue of CRC patients receiving surgical treatment in The the First Affiliated Hospital of Dali University from February 2019 to May 2020 were collected.Constructed CRC LoVo cells with overexpressing overexpression or knocking down of circ0091579 and miR-330-3 p were constructed.The expression of circ0091579,miR-330-3 p and RNF126 in CRC tissue and cell lines(HCT116,SW620,CW-2 and LoVo cells)was detected by qPCR.The proliferation,invasion and apoptosis was were measured by MTT assay,Transwell assay and flow Flow cytometry assay,respectively.The targeting relationship between circ0091579 and miR-330-3 p,miR-330-3 p and RNF126 was predicted by bioinformatics methodstool,and verified by dual Dual luciferase reporter experiments and RNA immunoprecipitation experiments.Results:In CRC tissues and cell lines,circ0091579 and RNF126 were both highly expressed,and miR-330-3 p was lowly expressed(all P<0.05).Knockdown of circ0091579 could inhibit the proliferation and invasion,but promote the apoptosis of CRC cells(P<0.05);however,this effect was reversed by miR-330-3 p-inhibitor.Overexpression of miR-330-3 p could suppress the ability of proliferation and invasion,but enhance the apoptosis of LoVo cells(P<0.05),and this effect was rescued after the participation of RNF126.There is a targeting relationship among circ0091579,miR-330-3 p and RNF126.Conclusion:circ0091579 promotes the proliferation,migration and invasion,inhibits the apoptosis of LoVo cells via miR-330-3 p/RNF126 axis.