Tumor Biology Laboratory, Department of Biochemistry and Molecular Biology, Chang Gung University College of Medicine, Gueishan, Taoyuan, Taiwan, Republic of China.
J Cell Physiol. 2014 Mar;229(3):309-22. doi: 10.1002/jcp.24448.
The oncogenic latent membrane protein 1 (LMP1) of Epstein-Barr virus (EBV) is involved in the pathogenesis of human nasopharyngeal carcinoma (NPC) and lymphoma. We and other authors have shown earlier that LMP1 induces apoptosis and inhibits xenograft tumor growth in mice, but the mechanism underlying these processes has not been investigated so far. In the present study, we show that knockdown of LMP1 renders the EBV-positive NPC cell line CG-1 resistant to various genotoxic drugs (cisplatin, etoposide, and adriamycin). LMP1 inhibits the expression of Cabin1, a Ca(2+) regulated protein shown earlier to inhibit calcineurin. Knockdown of calcineurin binding protein (Cabin1) with small hairpin RNA sensitizes CG-1 cells to genotoxic drugs. In contrast, LMP1 overexpression reduces Cabin1 level and renders both CG-1 cells and EBV-negative NPC cell lines sensitive to cisplatin. The c-Jun-N-terminal kinase (JNK) and ERK pathways are required for LMP1-induced suppression of Cabin1 at the transcriptional level. Chromatin immunoprecipitation assays further confirm that the JNK-activated transcription factor AP-1 mediates the LMP1-induced down-regulation of Cabin1 gene expression. LMP1 knockdown also increases the resistance of xenograph tumors to cisplatin in mice, therefore confirming the relevance of our findings in vivo. This study reveals the molecular mechanism underlying the pro-apoptotic activity of LMP1 during cisplatin-based NPC chemotherapy.
EBV 致癌潜伏膜蛋白 1(LMP1)参与人类鼻咽癌(NPC)和淋巴瘤的发病机制。我们和其他作者之前已经表明,LMP1 诱导凋亡并抑制小鼠异种移植肿瘤生长,但迄今为止尚未研究这些过程的机制。在本研究中,我们表明敲低 LMP1 使 EBV 阳性 NPC 细胞系 CG-1 对各种遗传毒性药物(顺铂、依托泊苷和阿霉素)具有抗性。LMP1 抑制 Cabin1 的表达,Cabin1 是一种先前显示抑制钙调神经磷酸酶的 Ca(2+) 调节蛋白。用短发夹 RNA 敲低钙调神经磷酸酶结合蛋白(Cabin1)可使 CG-1 细胞对遗传毒性药物敏感。相比之下,LMP1 过表达降低 Cabin1 水平,并使 CG-1 细胞和 EBV 阴性 NPC 细胞系对顺铂敏感。LMP1 诱导 Cabin1 在转录水平的抑制需要 c-Jun-N-末端激酶(JNK)和 ERK 途径。染色质免疫沉淀分析进一步证实,JNK 激活的转录因子 AP-1 介导 LMP1 诱导的 Cabin1 基因表达下调。LMP1 敲低也增加了异种移植肿瘤对顺铂的耐药性,因此证实了我们体内发现的相关性。这项研究揭示了 LMP1 在基于顺铂的 NPC 化疗中促凋亡活性的分子机制。