摘要
目的探讨circVRK1/miR-18b-5p轴对结直肠癌细胞增殖、迁移和凋亡的影响。方法实时定量聚合酶链式反应检测53例结直肠癌患者癌组织和癌旁组织中circVRK1和miR-18b-5p表达。体外培养结直肠癌LoVo细胞,双荧光素酶报告基因实验验证circVRK1与miR-18b-5p的调控关系。分别转染pcDNA-circVRK1、miR-18b-5p inhibitor,或共转染pcDNA-circVRK1与miR-18b-5p mimic至LoVo细胞,采用CCK-8法、克隆形成实验、划痕实验、流式细胞术、蛋白质印迹法分别检测细胞增殖、迁移、凋亡、蛋白(cleaved-caspase3、Bax、Bcl-2)表达。结果结直肠癌组织中的circVRK1表达量显著低于癌旁组织(P<0.05),miR-18b-5p表达量显著高于癌旁组织(P<0.05)。circVRK1可靶向结合miR-18b-5p,且过表达circVRK1抑制LoVo细胞中miR-18b-5p的表达。过表达circVRK1或敲减miR-18b-5p后,LoVo细胞的抑制率、凋亡率、cleaved-caspase3、Bax蛋白表达升高(P<0.05),克隆形成数、划痕愈合率、Bcl-2蛋白表达降低(P<0.05)。过表达miR-18b-5p导致过表达circVRK1对LoVo细胞增殖、迁移和凋亡的影响降低。结论结直肠癌组织中circVRK1呈低表达;过表达circVRK1可通过靶向抑制miR-18b-5p降低结直肠癌细胞的增殖和迁移能力,并加剧结直肠癌细胞凋亡。
Objective To investigate the effect of circVRK1 on the proliferation,migration and apoptosis of colorectal cancer cells.Methods The expression of circVRK1 and miR-18b-5p in the cancerous tissues and adjacent tissues of 53 patients with colorectal cancer were detected by real-time quantitative polymerase chain reaction.Colorectal cancer cells LoVo were cultured in vitro,and the regulatory relationship between circVRK1 and miR-18b-5p was verified by the dual luciferase reporter gene experiment.pcDNA-circVRK1 or miR-18b-5p inhibitor was transfected into LoVo cells,or pcDNA-circVRK1 and miR-18b-5p mimic were co-transfected into LoVo cells,and then cell proliferation,migration,apoptosis and the expression of proteins(cleaved-caspase3,Bax,Bcl-2)were detected by cell counting kit-8(CCK-8)assay,clone formation assay,scratch assay,flow cytometry,and Western blotting,respectively.Results The expression of circVRK1 in colorectal cancer tissue was lower than that in adjacent tissues(P<0.05),but the expression of miR-18b-5p was higher than that in adjacent tissues(P<0.05).CircVRK1 could target miR-18b-5p,and overexpression of circVRK1 inhibited the expression of miR-18b-5p in LoVo cells.After overexpression of circVRK1 or knockdown of miR-18b-5p,inhibition rate,apoptosis rate and the expression of cleaved-caspase3 and Bax protein were increased(P<0.05),but the number of clones,scratch healing rate and the expression of Bcl-2 protein were decreased(P<0.05).Overexpression of miR-18b-5p leaded to reduced effects of overexpression of circVRK1 on the proliferation,migration and apoptosis of LoVo cells.Conclusion CircVRK1 was under-expressed in colorectal cancer tissues;overexpression of circVRK1 could reduce the proliferation and migration of colorectal cancer cells LoVo by targeting miR-18b-5p,and increase the apoptosis of LoVo cells.
作者
应成龙
陈华斌
吴剑
YING Chenglong;CHEN Huabin;WU Jian(Department of General Surgery,Taizhou Integrated Traditional Chinese and Western Medicine Hospital,Taizhou 317500,Zhejiang,China)
出处
《中国现代医生》
2023年第1期60-66,共7页
China Modern Doctor
