沉默circ_0000218靶向miR-140-3p影响皮肤鳞状细胞癌细胞恶性生物学行为  

Silencing circ_0000218 Affects the Malignant Biological Behavior of Cutaneous Squamous Cell Carcinoma Cells by Targeting miR-140-3p

作　　者：李文静 肖钟[1] 王新[1] 何艳 Li Wenjing;Xiao Zhong;Wang Xin;He Yan(The First People's Hospital of Longquanyi District,Chengdu 610100,China)

机构地区：[1]成都市龙泉驿区第一人民医院,四川成都610100

出　　处：《中国中西医结合皮肤性病学杂志》2023年第1期1-7,共7页Chinese Journal of Dermatovenereology of Integrated Traditional and Western Medicine

基　　金：四川省卫生和计划生育委员会科研课题(编号:18PJ244)。

摘　　要：目的探讨circ_0000218对皮肤鳞状细胞癌(cSCC)细胞恶性生物学行为的影响及可能机制。方法收集21例cSCC组织和21例正常皮肤组织,实时荧光定量聚合酶链反应(RT-qPCR)法检测组织中circ_0000218和微小RNA-140-3p(miR-140-3p)表达。体外培养cSCC SCL-1细胞,分别转染si-circ_0000218、miR-140-3p mimics、共转染si-circ_0000218与anti-miR-140-3p后,采用细胞计数试剂盒-8(CCK-8)法和克隆形成实验检测细胞增殖,流式细胞术检测细胞凋亡,划痕实验检测细胞迁移,蛋白质印迹法(Western blotting)检测细胞中活化的胱天蛋白酶-3(Cleaved caspase-3)蛋白表达。双荧光素酶报告基因实验验证circ_0000218和miR-140-3p调控关系。结果cSCC组织中circ_0000218表达量高于正常皮肤组织(P<0.05),而miR-140-3p表达量低于正常皮肤组织(P<0.05);转染si-circ_0000218或miR-140-3p mimics后,cSCC SCL-1细胞光密度(OD)值、集落形成数及划痕愈合率降低(P<0.05),而凋亡率和细胞中Cleaved caspase-3蛋白表达量升高(P<0.05)。circ_0000218靶向负调控miR-140-3p。与转染si-circ_0000218的SCL-1细胞比较,共转染si-circ_0000218与anti-miR-140-3p的SCL-1细胞OD值、集落形成数及划痕愈合率升高(P<0.05),而凋亡率和细胞中Cleaved caspase-3蛋白表达量降低(P<0.05)。结论circ_0000218在cSCC组织中呈高表达,其可能通过靶向抑制miR-140-3p促进cSCC细胞增殖和迁移,而阻碍细胞凋亡,有可能成为cSCC治疗的分子靶点。Objective In order to investigate the effects of circ_0000218 on the malignant biological behavior of cutaneous squamous cell carcinoma(cSCC)cells and its possible mechanism.Methods Twenty-one cases of cSCC tissues and twenty-one cases of normal skin tissues were collected,and the expressions of circ_0000218 and microRNA-140-3p(miR-140-3p)in the tissues were detected by reverse transcription quantitative polymerase chain reaction(RT-qPCR).cSCC SCL-1 cells were cultured in vitro and respectively transfected with si-circ_0000218,miR-140-3p mimics,or co-transfected with si-circ_0000218 and anti-miR-140-3p.Then cell proliferation was detected by cell counting kit-8(CCK-8)and clone formation experiment;flow cytometry was used to detect cell apoptosis;scratch test was used to detect cell migration;Western blotting was used to detect the protein expression of cleaved caspase-3 in cells.The dual luciferase reporter gene experiment verified the regulatory relationship between circ_0000218 and miR-140-3p.Results The expression of circ_0000218 in cSCC tissue was higher than that in normal skin tissue(P<0.05),while the expression of miR-140-3p was lower than normal skin tissue(P<0.05).After cSCC SCL-1 cells were transfected with si-circ_0000218 or miR-140-3p mimics,the cell optical density(OD)value,colony formation number and scratch healing rate decreased(P<0.05),while apoptosis rate and the protein expression of cleaved caspase-3 in cells increased(P<0.05).circ_0000218 negatively regulated the expression of miR-140-3p.Compared with SCL-1 cells transfected with si-circ_0000218,SCL-1 cells co-transfected with si-circ_0000218 and anti-miR-140-3p had higher OD value,colony formation number and scratch healing rate(P<0.05),but had a lower apoptosis rate and the protein level of cleaved caspase-3 reduced(P<0.05).Conclusion circ_0000218 is highly expressed in cSCCtissues. It may promote the proliferation and migration of cSCC cells by targeting the expression of miR-140-3p,and hinder their apoptosis. It may become a molecular tar

关 键 词：皮肤鳞状细胞癌 circ_0000218 微小RNA-140-3p 细胞增殖 凋亡 迁移 

分 类 号：R739.5[医药卫生—肿瘤]