Chen Xinyan, Chen Yumei, Lin Xueyan, Su Shan, Hou Xiaoman, Zhang Qian, Tian Yongjie
Department of Obstetrics and Gynecology, Shandong Provincial Hospital Affiliated to Shandong University, No. 324 Jingwu Road, Jinan 250021, China.
Department of Gynecology and Obstetrics, Wenzhou People's Hospital, No. 57 Canghou Street, Wenzhou 325000, China.
Curr Pharm Biotechnol. 2018;19(6):506-513. doi: 10.2174/1389201019666180713102656.
BACKGROUND & OBJECTIVE: Many targeted ovarian cancer patients are resistant to olaparib treatment. Here we seek to understand the underlying molecular events and search for potential combinational therapeutics to surmount the intrinsic olaparib resistance in human ovarian cancer.
The cytotoxicity was determined by the MTT assay and cell viability was measured using Cell Counting Kit-8 (CCK-8). Protein expressions of ERK, P38, JNK, ERK5, LC3, N-CADHERIN, α-SMA were determined by western blotting. The invasion capacity was evaluated by the transwell chamber. Autophagy flux was monitored by the LC3 puncta formation. The epithelial-mesenchymal transition (EMT) markers were profiled by immunoblotting detection. The in vivo tumor progression was determined by xenograft mice model.
The olaparib-resistant cell lines were successfully generated in both SKOV3 and A2780 cells. The proliferative index was significantly higher in resistant cells in comparison with sensitive counterparts in the presence of olaparib. Both P38 and JNK were up-regulated in olaparib-resistant cells. The combinational treatment with P38-specific inhibitor SB202190 and JUN-specific inhibitor SP600125 significantly suppressed cell growth and migration, which was further attributed to the induction of autophagy flux and inhibition of EMT processing. We further consolidated the anti-tumor activities of SB202190 and SP600125 in xenograft mice.
Our data suggested that aberrant over-expression of P38 and JNK is causally linked to the olaparib resistance in ovarian cancer. Combination of P38 and JUN inhibitors demonstrated significant anti-tumor activity both in vitro and in vivo. Our study highlighted the potential therapeutic value of Mitogen-Activated Protein Kinase (MAPK) inhibitors in olaparib-resistant human ovarian cancer.
许多靶向治疗的卵巢癌患者对奥拉帕尼治疗耐药。在此，我们试图了解潜在的分子事件，并寻找潜在的联合疗法以克服人卵巢癌中固有的奥拉帕尼耐药性。
通过MTT法测定细胞毒性，使用细胞计数试剂盒-8（CCK-8）测量细胞活力。通过蛋白质印迹法测定ERK、P38、JNK、ERK5、LC3、N-钙黏蛋白、α-平滑肌肌动蛋白的蛋白表达。通过Transwell小室评估侵袭能力。通过LC3斑点形成监测自噬通量。通过免疫印迹检测分析上皮-间质转化（EMT）标志物。通过异种移植小鼠模型确定体内肿瘤进展。
在SKOV3和A2780细胞中均成功构建了奥拉帕尼耐药细胞系。在存在奥拉帕尼的情况下，耐药细胞的增殖指数明显高于敏感细胞。P38和JNK在奥拉帕尼耐药细胞中均上调。P38特异性抑制剂SB202190和JUN特异性抑制剂SP600125的联合治疗显著抑制细胞生长和迁移，这进一步归因于自噬通量的诱导和EMT过程的抑制。我们进一步在异种移植小鼠中证实了SB202190和SP600125的抗肿瘤活性。
我们的数据表明，P38和JNK的异常过表达与卵巢癌中的奥拉帕尼耐药性存在因果关系。P38和JUN抑制剂的联合在体外和体内均表现出显著的抗肿瘤活性。我们的研究突出了丝裂原活化蛋白激酶（MAPK）抑制剂在奥拉帕尼耐药的人卵巢癌中的潜在治疗价值。
