VAMP8促进人类胶质瘤细胞的细胞增殖和替莫唑胺耐药性。
VAMP8 facilitates cellular proliferation and temozolomide resistance in human glioma cells.
作者信息
Chen Yuanyuan, Meng Delong, Wang Huibo, Sun Ruochuan, Wang Dongrui, Wang Shuai, Fan Jiajun, Zhao Yingjie, Wang Jingkun, Yang Song, Huai Cong, Song Xiao, Qin Rong, Xu Tao, Yun Dapeng, Hu Lingna, Yang Jingmin, Zhang Xiaotian, Chen Haoming, Chen Juxiang, Chen Hongyan, Lu Daru
机构信息
State Key Laboratory of Genetic Engineering and MOE Key Laboratory of Contemporary Anthropology, School of Life Sciences, Shanghai, China (Y.C., D.M., D.W., Y.Z., J.W., C.H., X.S., D.Y., L.H., J.Y., H.C., H.C., D.L.); Department of Biosynthesis, School of Pharmacy, Fudan University, Shanghai, China (J.F.); Department of Neurosurgery, (H.W.); Department of Hematology, First Affiliated Hospital of Nanjing Medical University, Nanjing, China (S.W.); Eighth Department of General Surgery and Department of Pathology, First Affiliated Hospital of Anhui Medical University, Hefei, China (R.S., S.Y.); Department of Molecular Human Genetics, Baylor College of Medicine, Houston, Texas (X.Z.); Neurosurgery Research Institution of Shanghai, Department of Neurosurgery, Changzheng Hospital, Second Military Medical University, Shanghai, China (R.Q., T.X., J.C.).
出版信息
Neuro Oncol. 2015 Mar;17(3):407-18. doi: 10.1093/neuonc/nou219. Epub 2014 Sep 10.
BACKGROUND
Malignant glioma is a common and lethal primary brain tumor in adults. Here we identified a novel oncoprotein, vesicle-associated membrane protein 8 (VAMP8), and investigated its roles in tumorigenisis and chemoresistance in glioma.
METHODS
The expression of gene and protein were determined by quantitative PCR and Western blot, respectively. Histological analysis of 282 glioma samples and 12 normal controls was performed by Pearson's chi-squared test. Survival analysis was performed using the log-rank test and Cox proportional hazards regression. Cell proliferation and cytotoxicity assay were conducted using Cell Counting Kit-8. Autophagy was detected by confocal microscopy and Western blot.
RESULTS
VAMP8 was significantly overexpressed in human glioma specimens and could become a potential novel prognostic and treatment-predictive marker for glioma patients. Overexpression of VAMP8 promoted cell proliferation in vitro and in vivo, whereas knockdown of VAMP8 attenuated glioma growth by arresting cell cycle in the G0/G1 phase. Moreover, VAMP8 contributed to temozolomide (TMZ) resistance by elevating the expression levels of autophagy proteins and the number of autophagosomes. Further inhibition of autophagy via siRNA-mediated knockdown of autophagy-related gene 5 (ATG5) or syntaxin 17 (STX17) reversed TMZ resistance in VAMP8-overexpressing cells, while silencing of VAMP8 impaired the autophagic flux and alleviated TMZ resistance in glioma cells.
CONCLUSION
Our findings identified VAMP8 as a novel oncogene by promoting cell proliferation and therapeutic resistance in glioma. Targeting VAMP8 may serve as a potential therapeutic regimen for the treatment of glioma.
背景
恶性胶质瘤是成人常见的致命性原发性脑肿瘤。在此，我们鉴定出一种新型癌蛋白，即囊泡相关膜蛋白8（VAMP8），并研究了其在胶质瘤发生和化疗耐药中的作用。
方法
分别通过定量PCR和蛋白质印迹法检测基因和蛋白质的表达。采用Pearson卡方检验对282例胶质瘤样本和12例正常对照进行组织学分析。使用对数秩检验和Cox比例风险回归进行生存分析。使用细胞计数试剂盒-8进行细胞增殖和细胞毒性测定。通过共聚焦显微镜和蛋白质印迹法检测自噬。
结果
VAMP8在人类胶质瘤标本中显著过表达，可成为胶质瘤患者潜在的新型预后和治疗预测标志物。VAMP8的过表达在体外和体内均促进细胞增殖，而敲低VAMP8则通过使细胞周期停滞在G0/G1期来减弱胶质瘤生长。此外，VAMP8通过提高自噬蛋白的表达水平和自噬体数量导致对替莫唑胺（TMZ）耐药。通过小干扰RNA介导敲低自噬相关基因5（ATG5）或 syntaxin 17（STX17）进一步抑制自噬可逆转VAMP8过表达细胞中的TMZ耐药，而沉默VAMP8则损害胶质瘤细胞的自噬流并减轻TMZ耐药。
结论
我们的研究结果通过促进胶质瘤细胞增殖和治疗耐药确定VAMP8为一种新型癌基因。靶向VAMP8可能成为治疗胶质瘤的潜在治疗方案。
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