摘要
目的探讨circ_0013958靶向miR-532-3p在结直肠癌进展中的作用。方法RT-qPCR分析circ_0013958和miR-532-3p在结直肠癌组织和癌旁组织中的表达。用脂质体转染法分别将si-NC、si-circ_0013958、pcDNA、pcDNA-circ_0013958、miR-NC、mi R-532-3p mimics、si-circ_0013958+anti-miR-NC、si-circ_0013958+anti-miR-532-3p转染SW620细胞,通过CCK-8法和平板克隆实验评估细胞活力和克隆能力,通过划痕愈合和Transwell侵袭实验估细胞迁移和侵袭能力。利用Circular RNA Interactome网站预测circ_0013958与miR-532-3p的关系,并通过双荧光素酶报告基因法进一步验证。结果与癌旁组织相比,结直肠癌组织中circ_0013958表达上调,miR-532-3p表达下调。转染si-circ_0013958可上调miR-532-3p表达,降低细胞活力和划痕愈合率,并减少克隆形成数和侵袭力。转染pcDNA-circ_0013958可下调miR-532-3p表达;转染miR-532-3p mimics可降低细胞活力和划痕愈合率,并减少克隆形成数和侵袭力。circ_0013958和miR-532-3p直接结合。转染anti-miR-532-3p可显著减弱转染si-circ_0013958对SW620细胞活力、划痕愈合率、克隆形成数、侵袭数的影响。结论干扰circ_0013958通过促进miR-532-3p表达来抑制结直肠癌细胞增殖、迁移侵袭,从而抑制结直肠癌进展。
Objective To study the function of circ_0013958 targeting miR-532-3p in the progression of colorectal cancer.Methods The expression of circ_0013958 and miR-532-3p in colorectal cancer tissues and adjacent tissues was examined by RT-qPCR.si-NC,si-circ_0013958,pcDNA,pcDNA-circ_0013958,miR-NC,miR-532-3p mimics,si-circ_0013958+anti-miR-NC,si-circ_0013958+anti-miR-532-3p were respectively transfected into SW620 cells,and cell viability and cloning ability were evaluated by CCK-8 method and plate cloning assay,and abilities of cell migration and invasion were detected by scratch healing and Transwell assays.Circular RNA Interactome website was used to predict the interaction between circ_0013958 and miR-532-3p,which was further confirmed by Dual-Luciferase reporter assay.Results circ_0013958 was up-regulated,while miR-532-3p was down-regulated in colorectal cancer tissues.Transfection of si-circ_0013958 could up-regulate miR-532-3p expression,reduce cell viability and scratch healing rate,and reduce the number of clone formation and invasion.Transfection of pcDNA-circ_0013958 could down-regulate the expression of miR-532-3p.Transfection of miR-532-3p mimics could reduce cell viability and scratch healing rate,and reduce the number of clone formation and invasion.circ_0013958 directly binds to miR-532-3p.Transfection of anti-miR-532-3p could significantly reduce the effect of transfection of si-circ_0013958 on SW620 cell viability,scratch healing rate,number of clone formation,and number of invasion.Conclusion Interference circ_0013958 inhibits the proliferation,migration and invasion of colorectal cancer cells by promoting miR-532-3p expression,thereby inhibiting the progression of colorectal cancer.
作者
秦丽娟
王凡
贺小艳
Qin Lijuan;Wang Fan;He xiaoyan(Pharmaceutical Department,Chongqing university Jiangjin hospital,Chongqing 402260,China;Oncology department,Chongqing university Jiangjin hospital,Chongqing 402260,China)
出处
《中国组织化学与细胞化学杂志》
CAS
CSCD
2023年第3期236-244,共9页
Chinese Journal of Histochemistry and Cytochemistry
基金
重庆市江津区科委科技攻关项目Y2019046。
