TACSTD2调控食管鳞癌细胞对顺铂的敏感性

The regulation of TACSTD2 in the chemosensitivity of human esophageal squamous cell carcinoma

目的:观察干扰肿瘤关联钙信号转导因子2(tumor-associated calcium signal transducer 2,TACSTD2)表达对食管鳞癌细胞顺铂敏感性的影响及机制。方法:通过组织芯片检测食管鳞癌及癌旁组织中TACSTD2表达;选用人食管鳞癌细胞ECA 109、KYSE 30及其分别对应的顺铂(cisplatin,DDP)耐药株ECA 109/DDP、KYSE 30/DDP细胞,结合TACSTD2干扰技术,通过荧光定量PCR及蛋白质印迹法检测TACSTD2 mRNA和蛋白表达;CCK8实验和流式细胞术检测顺铂半抑制浓度(half maximal inhibitory concentration,IC50)以及细胞凋亡变化;蛋白质印迹法分析4种细胞中TACSTD2、多药耐药相关蛋白1(multidrug resistance protein 1,MDR1)、丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)及p-MAPK蛋白表达。结果:食管鳞癌中TACSTD2表达强度高于癌旁组织(P <0. 01);食管鳞癌顺铂耐药株中TACSTD2表达水平高于对应的食管鳞癌细胞表达(P均<0. 01);干扰TACSTD2后可以显著下调4种细胞中TACSTD2表达及IC50值(P均<0. 01);结合1μg/m L顺铂处理,TACSTD2干扰后4种细胞凋亡率明显增加(P均<0. 01);靶向下调TACSTD2后可见4种细胞中MAPK信号通路的活化抑制及MDR1表达显著降低。结论:靶向干预TACSTD2可提高食管鳞癌的顺铂敏感性,其作用机制可能与MAPK信号活化以及MDR1表达抑制有关。

Objective: To investigate the effects of tumor-associated calcium signal transducer 2( TACSTD2) on cisplatin of esophageal squamous cell carcinoma( ESCC) and the underlying mechanism. Methods: ESCC tissues and adjacent para-carcinoma tissues were collected for chip and IHC analysis. Choosing ESCC cells ECA 109,KYSE 30 and cisplatin( DDP) resistant cells ECA 109/DDP and KYSE 30/DDP as object treated with TACSTD2-siRNA,negative control-siRNA and non-treatment,the expression of TACSTD2 were detected by RT-PCR and Western blotting. CCK8 assay and flow cytometry were used to detect the DDP IC50 and the apoptosis of these cells. The expression of TACSTD2,multidrug resistance protein 1( MDR1),mitogen-activated protein kinase( MAPK) and p-MAPK proteins were detected by Western blotting. Results: The expression of TACSTD2 in ESCC tumors were higher than those of adjacent para-carcinoma tissues. After transfection with siRNA-TACSTD2,the relative expression of TACSTD2 and IC50 decreased more significantly than those of negative control group( P < 0. 01). With the treatment of cisplatin,TACSTD2-knockdown cells showed significantly higher apoptotic rate. The protein expression of p-MAPK and MDR1 in TACSTD2 transfected groups reduced significantly. Conclusion: Knockdown of TACSTD2 could make ESCC cells more sensitive to cisplatin,which is related to the downregulation of MAPK/MDR1 signaling pathway.