Pan Jie-Xue, Qu Fan, Wang Fang-Fang, Xu Jian, Mu Liang-Shan, Ye Long-Yun, Li Jun-Jian
The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325000, Zhejiang, China.
Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, 310006, Zhejiang, China.
Arch Gynecol Obstet. 2017 Dec;296(6):1145-1152. doi: 10.1007/s00404-017-4547-x. Epub 2017 Oct 3.
Resistance to platinum-based therapeutic agents is the major contributor to epithelial ovarian cancer (EOC) mortality. There is an urgent need to better understand the underlying mechanisms. Here we investigated the role of serpins in EOC chemoresistance and related mechanisms, and found that SERPINE1 played an important role in chemoresistance in A2780cp cells.
A2780cp and A2780s cells were used in our study. Microarray screening was used to identify the gene expression change under carboplatin treatment. A cell-counting kit-8 was used to detect the sensitivity of ovarian cancer cells after treatment. The expression of SERPINE1 was silenced by siRNA. The levels of SERPINE1 and epithelial-mesenchymal transition (EMT)-related proteins were confirmed by Western blot. MassArray EpiTYPER quantitative DNA methylation analysis was introduced to evaluate the methylation of the promoter of SERPINE1.
Microarray data showed that SERPINE1 and SERPINE2 increased most dramatically under carboplatin treatment in A2780cp cells. Carboplatin treatment could significantly increase the expression of SERPINE1 and induce the EMT process, with decreased expression of E-cadherin and increased expression of Vimentin, Snail and Twist. Knockdown of SERPINE1, but not SERPINE2, in A2780cp cells could inhibit the EMT process. We also found that hypomethylation in the promoter of SERPINE1 might result in the increased expression of SERPINE1 and subsequent EMT process in A2780cp cells.
Our study suggested that SERPINE1 may be a promising therapeutic target for chemoresistance.
对铂类治疗药物的耐药性是上皮性卵巢癌（EOC）死亡的主要原因。迫切需要更好地了解其潜在机制。在此，我们研究了丝氨酸蛋白酶抑制剂（serpins）在上皮性卵巢癌化疗耐药中的作用及相关机制，发现丝氨酸蛋白酶抑制剂E1（SERPINE1）在A2780cp细胞的化疗耐药中起重要作用。
本研究使用A2780cp和A2780s细胞。采用基因芯片筛选来确定卡铂处理下的基因表达变化。使用细胞计数试剂盒-8检测处理后卵巢癌细胞的敏感性。通过小干扰RNA（siRNA）使SERPINE1的表达沉默。通过蛋白质免疫印迹法确认SERPINE1和上皮-间质转化（EMT）相关蛋白的水平。引入MassArray EpiTYPER定量DNA甲基化分析来评估SERPINE1启动子的甲基化情况。
基因芯片数据显示，在A2780cp细胞中，卡铂处理下SERPINE1和丝氨酸蛋白酶抑制剂E2（SERPINE2）的表达增加最为显著。卡铂处理可显著增加SERPINE1的表达并诱导EMT过程，其中E-钙黏蛋白表达降低，波形蛋白、蜗牛蛋白和Twist蛋白表达增加。在A2780cp细胞中敲低SERPINE1而非SERPINE2可抑制EMT过程。我们还发现，SERPINE1启动子的低甲基化可能导致A2780cp细胞中SERPINE1表达增加及随后的EMT过程。
我们的研究表明，SERPINE1可能是化疗耐药的一个有前景的治疗靶点。
