摘要
目的探讨松果菊苷对皮肤鳞状细胞癌A431细胞增殖及凋亡的影响及其作用机制。方法体外培养人皮肤鳞状细胞癌A431细胞,采用不同质量浓度(12.5、25.0、50.0μg/L)的松果菊苷处理A431细胞。将circ_0046264对照空载体(pcDNA)、circ_0046264过表达载体(pcDNA-circ_0046264)、miR-510寡核苷酸阴性对照(anti-miR-NC)、miR-510寡核苷酸抑制物(miR-510 inhibitor)分别转染至A431细胞。将circ_0046264小分子干扰RNA(si-circ_0046264)、miR-510寡核苷酸模拟物(miR-510 mimic)分别转染至A431细胞后加入50.0μg/mL松果菊苷进行处理。采用MTT法、平板克隆形成实验与流式细胞术分别检测细胞增殖、克隆形成及凋亡率;qRT-PCR法与Western blotting法分别检测circ_0046264、miR-510、阿片样物质结合蛋白(OPCML)的表达量;双荧光素酶报告实验检测circ_0046264与miR-510的靶向关系,以及miR-510与OPCML的靶向关系。结果松果菊苷可浓度相关性地升高A431细胞增殖抑制率、凋亡率和circ_0046264、OPCML的表达量,并可降低miR-510的表达量,集落形成数减少(P<0.05)。转染pcDNA-circ_0046264或miR-510 inhibitor后,OPCML蛋白表达显著升高,细胞增殖抑制率和凋亡率显著升高,集落形成数明显减少(P<0.05)。circ_0046264可靶向调控miR-510的表达,OPCML是miR-510的靶基因;转染si-circ_0046264或miR-510 mimic均可降低松果菊苷对A431细胞增殖及凋亡的作用。结论松果菊苷可通过调节circ_0046264/miR-510/OPCML表达而抑制皮肤鳞状细胞癌细胞增殖及诱导细胞凋亡。
Objective To investigate the effects of echinoside on proliferation and apoptosis of squamous cell carcinoma A431 cells and its mechanism.Methods Human squamous cell carcinoma A431 cells were cultured in vitro and treated with echinoside at different concentrations(12.5,25.0,and 50.0μg/L).pcDNA,pcDNA-circ_0046264,anti-miR-NC,and miR-510 inhibitor were transfected into A431 cells,respectively.si-circ_0046264 and miR-510 mimic were transfected into A431 cells and 50.0μg/mL ecsinoside was added.MTT assay,plate clonogenesis assay,and flow cytometry were used to detect cell proliferation,clonogenesis and apoptosis rates,respectively.The expression levels of circ_0046264,miR-510,and OPCML were detected by qRT-PCR and Western blotting method,respectively.The targeting relationship between circ_0046264 and miR-510,and the targeting relationship between miR-510 and OPCML were detected by dual-lucifase reporting assay.Results Ecinosin increased proliferation inhibition and apoptosis rate,circ_0046264 and OPCML expression,decreased miR-510 expression level,and decreased colony formation number(P<0.05).After transfection with pcDNA-circ_0046264 or miR-510 inhibitor,OPCML protein expression,cell proliferation inhibition rate,and apoptosis rate were significantly increased,and colony formation number was significantly decreased(P<0.05).circ_0046264 can regulate the expression of miR-510,and OPCML is the target gene of miR-510.Transfection with si-circ_0046264 or miR-510 mimic could reduce the proliferation and apoptosis of A431 cells.Conclusion Echinoside can inhibit proliferation and induce apoptosis of squamous cell carcinoma cells by regulating circ_0046264/miR-510/OPCML expression.
作者
王康民
刘学伟
王秀菊
时晓玉
于小璇
WANG Kang-min;LIU Xue-wei;WANG Xiu-ju;SHI Xiao-yu;YU Xiao-xuan(Department of Dermatology,Zhengzhou Yihe Hospital,Zhengzhou 450000,China;Department of Dermatology,the First Affiliated Hospital of Henan University of Traditional Chinese Medicine,Zhengzhou 450000,China)
出处
《现代药物与临床》
CAS
2022年第2期230-236,共7页
Drugs & Clinic
基金
河南省医学科技攻关计划项目(LHGJ20191162)。
