Guo Xian-Zhi, Ye Xiao-Lei, Xiao Wei-Zhong, Wei Xue-Ni, You Qing-Hua, Che Xiao-Hang, Cai Yan-Jun, Chen Fang, Yuan Hao, Liu Xiao-Jian, Yu Ming-Hua
Department of Medical Oncology, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Pudong, Shanghai 201399, P.R. China.
Drugs and Pharmacology Laboratory, Ningbo Institute of Medical Sciences, Ningbo, Zhejiang 315020, P.R. China.
Oncol Rep. 2015 Nov;34(5):2557-66. doi: 10.3892/or.2015.4240. Epub 2015 Sep 1.
Vacuole membrane protein 1 (VMP1) was recently found to be involved in the process of tumor metastasis and is also considered to play a vital role in balancing apoptosis and autophagy. In the present study, the expression of VMP1 in colorectal cancer and matched adjacent non‑cancerous tissues was evaluated by immunohistochemistry (IHC) for studying the role of VMP1 in the process of colorectal cancer. Kaplan‑Meier analysis and the log-rank test were used to calculate the correlation of classic clinicopathological characteristics related to survival and the expression of VMP1. In vitro, a VMP1 stable gene silencing cell model was constructed using a lentiviral vector. The invasive ability and proliferation of colorectal cancer cells were evaluated by Transwell and MTT assays, respectively, and the underlying signaling pathway was explored by western blotting. Additionally, drug susceptibility to cisplatin, oxaliplatin and 5-FU was tested before and after VMP1 knockout. Finally, an animal model was constructed to explore the role of VMP1 in the physiopathologic process of colorectal cancer. Our results indicated that VMP1 showed increased expression in the adjacent non-cancer tissues compared with that in the colorectal cancer tissues. For different stages of colorectal cancer, expression of VMP1 had a negative correlation with the malignancy of the cancer. In clinical research, we also found that the median survival of patients with low VMP1 expression was much shorter than the survival of patients with high expression. In vitro, after infection with the lentivirus, cells with VMP1 knockout gained significant aggressive properties in regards to invasion and proliferation, and the mechanisms may be related to the activation of the PI3K/Akt/ZO-1/E-cadherin pathway. We also found that shVMP1 cells were more sensitive to 5-FU, but not cisplatin and oxaliplatin. Finally, we found a higher number of formed nodules in nude mice after intraperitoneal injection with shVMP1 cells in the in vivo study.
液泡膜蛋白1（VMP1）最近被发现参与肿瘤转移过程，并且被认为在平衡细胞凋亡和自噬中起着至关重要的作用。在本研究中，通过免疫组织化学（IHC）评估VMP1在结直肠癌及配对的相邻非癌组织中的表达，以研究VMP1在结直肠癌发生过程中的作用。采用Kaplan-Meier分析和对数秩检验来计算与生存相关的经典临床病理特征与VMP1表达之间的相关性。在体外，使用慢病毒载体构建VMP1稳定基因沉默细胞模型。分别通过Transwell和MTT试验评估结直肠癌细胞的侵袭能力和增殖能力，并通过蛋白质印迹法探索潜在的信号通路。此外，在敲除VMP1前后测试对顺铂、奥沙利铂和5-氟尿嘧啶的药物敏感性。最后，构建动物模型以探索VMP1在结直肠癌病理生理过程中的作用。我们的结果表明，与结直肠癌组织相比，VMP1在相邻非癌组织中的表达增加。对于不同阶段的结直肠癌，VMP1的表达与癌症的恶性程度呈负相关。在临床研究中，我们还发现VMP1低表达患者的中位生存期远短于高表达患者的生存期。在体外，感染慢病毒后，敲除VMP1的细胞在侵袭和增殖方面具有显著的侵袭性，其机制可能与PI3K/Akt/ZO-1/E-钙黏蛋白途径的激活有关。我们还发现shVMP1细胞对5-氟尿嘧啶更敏感，但对顺铂和奥沙利铂不敏感。最后，在体内研究中，我们发现腹腔注射shVMP1细胞的裸鼠体内形成的结节数量更多。
