Chong Kowit Yu, Hsu Chih-Jung, Hung Tsai-Hsien, Hu Han-Shu, Huang Tsung-Teng, Wang Tzu-Hao, Wang Chihuei, Chen Chuan-Mu, Choo Kong Bung, Tseng Ching-Ping
a Department of Medical Biotechnology and Laboratory Science; College of Medicine ; Chang Gung University ; Tao-Yuan , Taiwan , Republic of China.
Cancer Biol Ther. 2015;16(1):149-59. doi: 10.4161/15384047.2014.987093.
Multiple drug resistance (MDR) is a major obstacle to attenuating the effectiveness of chemotherapy to many human malignancies. Proteasome inhibition induces apoptosis in a variety of cancer cells and is recognized as a novel anticancer therapy approach. Despite its success, some multiple myeloma patients are resistant or become refractory to ongoing treatment by bortezomib suggesting that chemoresistant cancer cells may have developed a novel mechanism directed against the proteasome inhibitor. The present study aimed to investigate potential mechanism(s) of attenuation in a MDR cell line, MES-SA/Dx5. We found that compared to the parental human uterus sarcoma cell line MES-SA cells, MES-SA/Dx5 cells highly expressed the ABCB1 was more resistant to MG132 and bortezomib, escaping the proteasome inhibitor-induced apoptosis pathway. The resistance was reversed by co-treatment of MG132 and the ABCB1 inhibitor verapamil. The data indicated that ABCB1 might play a role in the efflux of MG132 from the MES-SA/Dx5 cells to reduce MG132-induced apoptosis. Furthermore, the canonical Wnt pathway was found activated only in the MES-SA/Dx5 cells through active β-catenin and related transactivation activities. Western blot analysis demonstrated that Wnt-targeting genes, including c-Myc and cyclin D1, were upregulated and were relevant in inhibiting the expression of p21 in MES-SA/Dx5 cells. On the other hand, MES-SA cells expressed high levels of p21 and downregulated cyclin D1 and caused cell cycle arrest. Together, our study demonstrated the existence and participation of ABCB1 and the Wnt pathway in an MDR cell line that attenuated proteasome inhibitor-induced apoptosis.
多药耐药(MDR)是削弱化疗对许多人类恶性肿瘤疗效的主要障碍。蛋白酶体抑制可诱导多种癌细胞凋亡,被认为是一种新型抗癌治疗方法。尽管取得了成功,但一些多发性骨髓瘤患者对硼替佐米的持续治疗产生耐药或变得难治,这表明化疗耐药癌细胞可能已经形成了一种针对蛋白酶体抑制剂的新机制。本研究旨在探讨MDR细胞系MES-SA/Dx5中衰减的潜在机制。我们发现,与亲本人类子宫肉瘤细胞系MES-SA细胞相比,MES-SA/Dx5细胞高度表达ABCB1,对MG132和硼替佐米更具耐药性,逃避了蛋白酶体抑制剂诱导的凋亡途径。MG132与ABCB1抑制剂维拉帕米联合处理可逆转耐药性。数据表明,ABCB1可能在MG132从MES-SA/Dx5细胞中流出以减少MG132诱导的凋亡中发挥作用。此外,通过活性β-连环蛋白和相关反式激活活性发现经典Wnt通路仅在MES-SA/Dx5细胞中被激活。蛋白质印迹分析表明,包括c-Myc和细胞周期蛋白D1在内的Wnt靶向基因在MES-SA/Dx5细胞中上调,并且与抑制p21的表达相关。另一方面,MES-SA细胞表达高水平的p21并下调细胞周期蛋白D1,导致细胞周期停滞。总之,我们的研究证明了ABCB1和Wnt通路在一个衰减蛋白酶体抑制剂诱导凋亡的MDR细胞系中的存在和参与。