土贝母苷甲通过调控circ_0000376/miR-203轴影响皮肤鳞状细胞癌SCL-1细胞恶性表型  

Tubeimoside-1 affects the phenotype of skin squamous cell carcinoma SCL-1 cells by regulating the circ_0000376/miR-203 axis

作　　者：王娟 肖春才 屈小燕 张晨阳 WANG Juan;XIAO Chuncai;QU Xiaoyan;ZHANG Chenyang(Department of Dermatology,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou 450007,China)

机构地区：[1]郑州大学附属郑州中心医院皮肤科,河南郑州450007

出　　处：《皮肤性病诊疗学杂志》2021年第2期90-98,共9页Journal of Diagnosis and Therapy on Dermato-venereology

摘　　要：目的:探讨土贝母苷甲对皮肤鳞状细胞癌SCL-1细胞恶性表型的影响及可能机制。方法:将体外SCL-1细胞分为对照组、不同剂量(5、10、20μg/mL)土贝母苷甲组、si-NC组、si-circ_0000376组、土贝母苷甲+pcDNA组和土贝母苷甲+pcDNA-circ_0000376组,CCK-8法检测细胞增殖抑制率,Transwell检测细胞迁移和侵袭,流式细胞仪检测细胞凋亡,Western blot检测细胞中Ki-67、MMP-2、MMP-9、Bcl-2和Bax蛋白表达,RT-qPCR检测circ_0000376和miR-203表达。双荧光素酶报告基因实验验证circ_0000376和miR-203调控关系。对照组与不同剂量土贝母苷甲组各检测指标比较用单因素方差分析;si-NC组与si-circ_0000376及土贝母苷甲+pcDNA组与土贝母苷甲+pcDNA-circ_0000376组各检测指标比较行独立样本t检验。结果:与对照组比较,土贝母苷甲组SCL-1细胞增殖抑制率、凋亡率及Bax蛋白表达升高(F值分别为772.61、352.20、277.56,P值均<0.01),细胞迁移数、侵袭数及Ki-67、MMP-2、MMP-9和Bcl-2蛋白表达降低(F值分别为125.57、180.50、257.87、301.22、399.27、233.29,P值均<0.01),circ_0000376表达降低(F=205.36,P<0.01),miR-203表达升高(F=247.14,P<0.01),且呈剂量依赖性。与si-NC组比较,si-circ_0000376组SCL-1细胞增殖抑制率、凋亡率及Bax蛋白表达升高(t值分别为36.78、21.56、25.20,P值均<0.01),细胞迁移数、侵袭数及Ki-67、MMP-2、MMP-9和Bcl-2蛋白表达降低(t值分别为16.00、17.79、21.73、21.02、21.62、19.68,P值均<0.01)。双荧光素酶报告基因实验结果显示,circ_0000376靶向负调控miR-203。与土贝母苷甲+pcDNA组比较,土贝母苷甲+pcDNA-circ_0000376组SCL-1细胞增殖抑制率、凋亡率及Bax蛋白表达降低(t值分别为35.31、19.65、19.55,P值均<0.01),细胞迁移数、侵袭数及Ki-67、MMP-2、MMP-9和Bcl-2蛋白表达升高(t值分别为12.27、21.37、24.15、23.40、23.48、22.28,P值均<0.01)。结论:土贝母苷甲可能通过调控circ_0000376/miR-203轴抑制皮肤鳞状细Objective:To investigate the effect of tubeimoside-1 on the phenotype of skin squamous cell carcinoma SCL-1 cells and possible mechanism of action.Methods:SCL-1 cells were divided into the control group and the treatment groups,including different doses(5,10,20μg/mL)of tubeimoside-1 groups,si-NC group,si-circ_0000376 group,tubeimoside-1+pcDNA group,and tubeimoside-1+pcDNA-circ_0000376 group.CCK-8 method was used to detect cell proliferation inhibition rate.Transwell was used to detect cell migration and invasion.Flow cytometry was used to detect cell apoptosis.Western blot was used to detect the protein expression of Ki-67,MMP-2,MMP-9,Bcl-2 and Bax.RT-qPCR method was used to detect the expression of circ_0000376 and miR-203.The dual luciferase reporter gene experiment was used to verify the regulatory relationship between circ_0000376 and miR-203.Results:Compared with the control group,the proliferation inhibition rate,apoptosis rate and the Bax protein expression of cells in the tubeimoside-1 group were significantly increased(F=772.61,352.20,277.56,all P<0.01),while the number of cell migration and invasion,and Ki-67,MMP-2,MMP-9 and Bcl-2 protein expressions of cells in the tubeimoside-1 group were significantly decreased(F=125.57,180.50,257.87,301.22,399.27,233.29,all P<0.01).The expression of circ_0000376 of cells in the tubeimoside-1 group was significantly lower than that in the control group(F=205.36,P<0.01),while the expression of miR-203 of cells in the tubeimoside-1 group was significantly higher than that in the control group(F=247.14,P<0.01),and the relation was dose-dependent.Compared with the si-NC group,the cell proliferation inhibition rate,apoptosis rate and the Bax protein expression of cells in si-circ_0000376 groups were significantly increased(t=36.78,21.56,25.20,all P<0.01),while the number of migration and invasion,and Ki-67,MMP-2,MMP-9 and Bcl-2 protein expressions of cells in si-circ_0000376 groups were significantly decreased(t=16.00,17.79,21.73,21.02,21.62,19.68,all P<0.01).The results

关 键 词：土贝母苷甲 皮肤鳞状细胞癌 circ_0000376 miR-203 细胞增殖 迁移 侵袭 凋亡 

分 类 号：R739.5[医药卫生—肿瘤]