SOX4对非小细胞肺癌细胞A549的顺铂耐药作用的影响

Role of SOX4 on DDP Resistance in Non-small Cell Lung Cancer Cell of A549

背景与目的肺癌是最严重的恶性肿瘤之一,其中非小细胞肺癌发病率在肺癌中居首位。部分患者对顺铂耐受是导致化疗失败的主要原因。SOX4是转录调节因子,在多种肿瘤的发生发展过程中发挥着重要的作用,通过调控β-catenin的表达对Wnt信号通路进行调控。本研究旨在探讨SOX4对非小细胞肺癌细胞A549的顺铂耐药作用的影响。方法体外诱导法建立顺铂耐药的肺癌细胞株A549/DDP,CCK8法检测A549及A549/DDP细胞对顺铂的耐药性。绘制A549与A549/DDP细胞生长曲线。Western blot检测耐药细胞株中SOX4的蛋白表达水平。CCK8法检测敲减SOX4后耐药细胞株A549/DDP对顺铂耐药性。实时定量PCR和免疫印迹法检测敲减SOX4后A549/DPP细胞中β-catenin和Survivin蛋白的表达。结果成功构建非小细胞肺癌顺铂耐药细胞株A549/DDP,其耐药性是亲本细胞的13.7倍,差异有统计学意义(P<0.001)。生长曲线显示耐药细胞株与亲本细胞增殖速度没有统计学差异(P<0.05);与A549细胞相比,耐药细胞A549/DDP细胞中SOX4的表达显著增高(P<0.001)。敲减SOX4后,A549/DDP细胞的耐药性显著降低;敲减SOX4后,A549/DDP细胞中β-catenin和Survivin的m RNA和蛋白表达水平显著降低。结论 SOX4的表达水平影响非小细胞肺癌细胞A549对顺铂的耐药性。

Background and objective Lung cancer is one of the most serious disease and the incidence of non- small cell lung cancer （NSCLC） is the highest in lung cancer. The main reason for the failure of chemotherapy is the tolerance to cisplatin. Transcriptional regulator SOX4 plays an important role in the occurrence and development of many tumors, and regulates Wnt signaling pathway by regulating the expression of β-catenin. We aimed to investigate the role of SOX4 on cisplat- in-resistance in NSCLC cell A549 cell. Methods The cisplatin-resistance lung cancer cell line A549/DDP was constructed by induction method in vitro, and cisplatin-resistance detected by CCK8 assay. Growth curves ofA549 and A549/DDP was calculated. The expression level of SOX4 in A549 and A549/DDP cells were detected by Western blot. A549/DDP were knock- down of SOX4 by siRNA transfection, and the cisplatin-resistance of detected by CCK-8 assay the expression level of β-catenin and Survivin were detected by real-time PCR and Western blot. Results The cisplatin-resistance cell line A549/DDP was constructed successfully, and its cisplatin-resistance is 13.7 times higher than in A549. ＂lhere was no significance difference between A549 and A549/DDP in cell proliferation. The expression level of SOX4 is higher in A549/DDP than in A549. The cisplatin-resistance significantly decreased in A549/DDP cells after knockdown of SOX4 by siRNA transfection. The expres- sion level of β-catenin and Survivin significantly decreased in AS49/DDP cells after knockdown of SOX4. Conclusion SOX4 can strengthen cisplatin-resistance of non-small cell lung cancer cell A549.