TACSTD2调控食管鳞癌细胞对顺铂的敏感性

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1 μg/mL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。

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

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.