紫草素对肾癌OS-RC-2细胞生物学行为及RhoA/ROCK信号通路的影响  

Effects of shikonin on biological behavior of renal carcinoma OS-RC-2 cells and RhoA/ROCK signaling pathway

作　　者：韩坤岭[1] 平玉杰[1] 王俊红 李慧敏[1] 霍仲超 Han Kunling;Ping Yujie;Wang Junhong;Li Huimin;Huo Zhongchao(Department of Urology,Handan Central Hospital,Handan 056000,China)

机构地区：[1]邯郸市中心医院泌尿外一科,邯郸056000

出　　处：《国际泌尿系统杂志》2024年第6期1046-1050,共5页International Journal of Urology and Nephrology

基　　金：河北省医学科学研究课题计划项目(20200459)。

摘　　要：目的探讨紫草素对肾癌OS-RC-2细胞增殖、凋亡和迁移的影响及对Ras同源基因家族成员A(RhoA)/Rho相关卷曲螺旋蛋白激酶(ROCK)信号通路的调控作用。方法肾癌OS-RC-2细胞正常传代培养,取OS-RC-2细胞分为空白对照组(常规培养OS-RC-2细胞)、紫草素低浓度组(含10μg/mL的紫草素)、紫草素中浓度组(含20μg/mL的紫草素)、紫草素高浓度组(含40μg/mL的紫草素)和阳性对照组(含40μg/mL的顺铂),培养48 h后,采用CCK-8法检测OS-RC-2细胞的增殖率,流式细胞术检测OS-RC-2细胞的凋亡率,划痕试验法检测OS-RC-2细胞的迁移率,荧光定量PCR法检测OS-RC-2细胞中的RhoA、ROCK1、ROCK2的mRNA表达水平,蛋白印迹法检测OS-RC-2细胞中RhoA、ROCK1、ROCK2的蛋白表达水平。结果与空白对照组相比,紫草素低、中、高浓度组中OS-RC-2细胞的增殖率、迁移率降低,细胞凋亡率升高(均P<0.05)。与空白对照组相比,紫草素低、中、高浓度组OS-RC-2细胞中的RhoA、ROCK1、ROCK2 mRNA和蛋白表达水平降低(均P<0.05)。结论紫草素可有效抑制肾癌OS-RC-2细胞的增殖和迁移,促进其凋亡,其机制与抑制RhoA/ROCK信号通路的激活有关。ObjectiveTo investigate the effects of shikonin on the proliferation,apoptosis and migration of renal carcinoma OS-RC-2 cells and the regulation of Ras homolog gene family member A(RhoA)/Rho-associated coiled-coil protein kinase(ROCK)signaling pathway.MethodsThe enal carcinoma OS-RC-2 cells were cultured under normal conditions.The OS-RC-2 cells at logarithmic growth stage were divided into the following groups:blank control group(conventional culture of OS-RC-2 cells),low concentration group(containing 10μg/mL of shikonin),medium concentration group(containing 20μg/mL of shikonin),high concentration group(containing 40μg/mL of shikonin),and positive control group(containing 40μg/mL cisplatin).They were cultured for 48 hours,during which the proliferation rate of OS-RC-2 cells was determined using the CCK-8 method,the apoptosis rate was measured by flow cytometry,and cell mobility was assessed through a scratch test.Furthermore,the mRNA expression levels of RhoA,ROCK1,and ROCK2 in OS-RC-2 cells were quantified using fluorescence quantitative PCR,while their protein expression levels were analyzed via Western blot.ResultsCompared to the blank control group,the proliferation and mobility rates of OS-RC-2 cells in the low,medium,and high concentrations of shikonin groups exhibited a successive decrease,while the apoptosis rate showed a successive increase(all P<0.05).Furthermore,compared to the blank control group,both mRNA and protein expression levels of RhoA,ROCK1,and ROCK2 in OS-RC-2 cells decreased successively with increasing concentrations of shikonin(all P<0.05).ConclusionsThe compound shikosin effectively inhibits the proliferation and migration of renal carcinoma OS-RC-2 cells while promoting their apoptosis,potentially through the inhibition of RhoA/ROCK signaling pathway activation.

关 键 词：肾肿瘤 紫草素 RhoA/ROCK信号通路 细胞凋亡 

分 类 号：R737.11[医药卫生—肿瘤]