PDIA4在肾癌细胞舒尼替尼耐药中的作用研究  

Study on the Role of PDIA4 in Sunitinib Resistance of Renal Cancer Cells

作　　者：唐麒麟 柯波亮 何雏江 徐子杰 陈磊[1] 邵怡[1] TANG Qi-lin;KE Bo-liang;HE Chu-jiang;XU Zi-jie;CHEN Lei;SHAO Yi(Department of Urology,Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai,200080,China)

机构地区：[1]上海交通大学医学院附属第一人民医院泌尿外科,上海200080

出　　处：《现代生物医学进展》2024年第13期2408-2416,共9页Progress in Modern Biomedicine

基　　金：上海市科学技术委员会项目(23141902100)。

摘　　要：目的:建立肾癌舒尼替尼耐药细胞株,探讨蛋白二硫键异构酶A成员4(PDIA4)调控肾细胞癌舒尼替尼耐药的机制。方法:采用小剂量间歇诱导法建立786-O和OS-RC-2耐药细胞株(786-OR和OS-RC-2R),采用实时荧光定量PCR法、蛋白免疫印迹法检测PDIA4在耐药肾癌细胞株与非耐药肾癌细胞株中的表达情况,通过慢病毒感染构建PDIA4稳定过表达或敲减肾癌细胞系,通过细胞计数试剂盒(CCK-8)法检测PDIA4对肾癌细胞增殖影响;通过qRT-PCR和Western blot检测凋亡相关蛋白BCL2、BAX、Caspase3、Cleaved-Caspase3、Caspase9表达。结果:与对照组亲本细胞株786-O和OS-RC-2相比,耐药细胞株786-OR和OS-RC-2R的PDIA4表达水平明显升高(P<0.05)。过表达PDIA4可促进肾癌细胞的生长和活力以及降低肾癌细胞对舒尼替尼的敏感性(P<0.05);而敲低PDIA4能抑制肾癌细胞的生长和活力以及升高肾癌细胞对舒尼替尼的敏感性(P<0.05)。此外,过表达PDIA4能抑制促凋亡蛋白的表达(P<0.05),而敲低PDIA4能促进促凋亡蛋白的表达(P<0.05)。结论:PDIA4与肾细胞癌舒尼替尼耐药有关,其机制是抑制细胞凋亡引起肾癌细胞癌舒尼替尼耐药,靶向PDIA4则可以逆转耐药。Objective:To establish a renal cell carcinoma sunitinib-resistant cell line and to explore the mechanism by which protein disulfide isomerase A4(PDIA4)regulates sunitinib resistance in renal cell carcinoma.Methods:A small-dose intermittent induction method was used to establish 786-O and OS-RC-2 resistant cell lines(786-OR and OS-RC-2R),qRT-PCR and Western blot were used to detect the expression of PDIA4 in drug-resistant renal carcinoma cell lines and non-drug-resistant renal carcinoma cell lines,and lentiviral infection was used to construct a stable overexpression or knockdown renal carcinoma cell line of PDIA4.The effect of PDIA4 on the proliferation of renal cancer cells was detected by CCK-8 assay;the expression of apoptosis-related proteins BCL2,BAX,Caspase3,Cleaved-Caspase3,and Caspase9 was detected by RT-qPCR and Western blot.Results:Compared with the control group of parental cells786-O and OS-RC-2,the PDIA4 expression levels of drug-resistant cell lines 786-OR and OS-RC-2R are significantly increased(P<0.05).Overexpression of PDIA4 promoted the growth and viability of renal cancer cells as well as decreased the sensitivity of renal cancer cells to sunitinib(P<0.05),whereas knockdown of PDIA4 inhibited the growth and viability of renal cancer cells as well as elevated the sensitivity of renal cancer cells to sunitinib(P<0.05).In addition,overexpression of PDIA4 inhibited the expression of pro-apoptotic proteins(P<0.05),while knockdown of PDIA4 promoted the expression of pro-apoptotic proteins(P<0.05).Conclusions:PDIA4 is associated with sunitinib resistance in renal cell carcinoma by the mechanism of inhibition of apoptosis causing sunitinib resistance in renal cell carcinoma,and targeting PDIA4 reverses the resistance.

关 键 词：PDIA4 肾细胞癌 舒尼替尼耐药 凋亡 

分 类 号：R-33[医药卫生] R737.11