SOX4抑制凋亡介导食管癌顺铂耐药的实验研究

目的：探讨SOX4与食管癌顺铂耐药的关系及其耐药机制。方法：采用递增药物浓度、间歇冲击作用的方法构建顺铂食管癌耐药细胞株KYSE30/DDP；流式细胞仪检测KYSE30及KYSE30/DDP细胞凋亡情况；Western blot检测SOX4及凋亡相关指标在KYSE30及KYSE30/DDP细胞中的变化；CCK8检测干扰SOX4对KYSE30和KYSE30/DDP细胞顺铂敏感度的影响；流式细胞仪及Western blot检测干扰SOX4对KYSE30/DDP细胞凋亡及相关指标的影响。结果：成功构建了生长状态良好的KYSE30/DDP细胞株，其耐药指数为4.1；KYSE30/DDP细胞凋亡较其亲本细胞KYSE30明显降低（t=-8.414,P<0.05）；Western blot结果显示，SOX4在顺铂耐药细胞KYSE30/DDP中的表达高于其亲本细胞KYSE30，KYSE30/DDP细胞中凋亡相关指标Cleaved caspase 3及Cleaved PARP表达低于KYSE30；在KYSE30/DDP细胞中干扰SOX4，其对顺铂的敏感性及凋亡细胞增加（F=625.423,P<0.05），凋亡相关指标Cleaved caspase 3及Cleaved PARP表达增加。结论：SOX4在食管癌顺铂耐药细胞KYSE30/DDP中高表达，并可通过抑制凋亡介导食管癌顺铂耐药。

Experimental Study of SOX4 Mediated Cisplatin Resistance in Esophagus Cancer by Inhibiting Apoptosis

Objective: To explore the relationship between cisplatin resistance and the expression of SOX4,and related drug resistance mechanisms in esophagus cancer.Methods: The cisplatin resistance cell line KYSE30/DDP was established in culture by a program of treating KYSE30 parent cells in culture with increasing concentration of cisplatin.The apoptosis of KYSE30 and KYSE30/DDP cells were determined by flow cytometry.Western blot detected the expression of SOX4 and apoptosis related indicators in KYSE30 and KYSE30/DDP cells.The influence of drug sensitively by knockdown the expression of SOX4 in KYES30 and KYSE30/DDP cell was measured by CCK8 assay.The change of apoptosis and related indicators in KYSE30/DDP cell after knockdown the expression of SOX4 was evaluated by western blot and flow cytometry.Results: CCK8 assay showedthe cisplatin resistance cell line KYSE30/DDP was successfully built,and the resistance index was 4.1.The apoptosis of KYSE30/DDP cell was significantly decreased compared with KYSE30 parent cell(P<0.05).Western blot indicated the expression of SOX4 are higher in KYSE30/DDP cell compared to its parent cell.And the expression of apoptosis related indicators lower in KYSE30/DDP cell.Knock down the expression of SOX4 in KYSE30/DDP cellfacilitated thecisplatin sensitivity andpromoted cell apoptosis(P<0.05),with the increase expression of apoptosis related indicators.Conclusion: SOX4 was high-expression in cisplatin resistance cell KYSE30/DDP,and mediated cisplatin resistance by inhibiting apoptosis.