一种在人类脑胶质瘤中表达的新基因 RNF138 及其在 U251 脑胶质瘤细胞系中的功能。
A novel gene RNF138 expressed in human gliomas and its function in the glioma cell line U251.
机构信息
Department of Neurosurgery, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu Province, China.
出版信息
Anal Cell Pathol (Amst). 2012;35(3):167-78. doi: 10.3233/ACP-2011-0051.
BACKGROUND
The gliomas represent the most common primary malignant brain tumors; however, little is known about the molecular pathogenesis of these tumors. Recent research reveals that the oncogenesis and development of gliomas have a close relation to the overexpression of several oncogenes and the inactivation of tumor suppressor genes. Whether the RING finger protein, RNF138, a newly discovered protein, plays a role in glioma oncogenesis is unknown. The present study investigates the expression levels of RNF138 mRNA in glioma samples and noncancerous brain samples and its function in the human glioma cell line U251.
METHODS
RT-PCR was used to ascertain the expression of RNF138 mRNA in the glioma cell lines U251, SHG44, U87, A172, and U373. The RNF138 mRNA expression levels of 35 pathological confirmed glioma samples (Grade I - 4 cases, Grade II - 13 cases, Grade III - 11 cases, and Grade IV - 7 cases) and five noncancerous brain tissue samples were analyzed by real-time quantitative PCR. By RNA interference (RNAi) with the lentivirus vector system, the expression of RNF138 was inhibited in the human astrocytomas-glioblastoma multiforme cell line U251. The effects of RNF138-knockdown on cell proliferation were assessed by Cellomics, and cell cycle and cell apoptosis were assessed by FACS.
RESULTS
The RNF138 mRNA is expressed in the five glioma cell lines, and its expression level is significantly higher in glioma tissue than in noncancerous brain tissue. By down-regulation of RNF138 expression, U251 cell proliferation was inhibited and cell apoptosis increased. At the same time, S stage cells lessened and G2 stage cells increased.
CONCLUSION
The RNF138 gene is highly expressed in glioma tissue and glioma cell lines. It plays an important role in glioma cell proliferation, apoptosis, and cell cycle.
背景
神经胶质瘤是最常见的原发性恶性脑肿瘤，但人们对这些肿瘤的分子发病机制知之甚少。最近的研究表明，神经胶质瘤的发生和发展与几个癌基因的过度表达和肿瘤抑制基因的失活密切相关。新发现的蛋白质 RING 指蛋白 138（RNF138）是否在神经胶质瘤的发生中起作用尚不清楚。本研究检测了 RNF138mRNA 在神经胶质瘤样本和正常脑组织样本中的表达水平及其在人神经胶质瘤细胞系 U251 中的功能。
方法
采用 RT-PCR 法检测神经胶质瘤细胞系 U251、SHG44、U87、A172 和 U373 中 RNF138mRNA 的表达。采用实时定量 PCR 法检测 35 例经病理证实的神经胶质瘤样本（Ⅰ级 4 例、Ⅱ级 13 例、Ⅲ级 11 例、Ⅳ级 7 例）和 5 例正常脑组织样本的 RNF138mRNA 表达水平。利用慢病毒载体系统的 RNA 干扰（RNAi）抑制人星形胶质细胞瘤-多形性胶质母细胞瘤细胞系 U251 中 RNF138 的表达。通过 Cellomics 检测 RNF138 敲低对细胞增殖的影响，通过 FACS 检测细胞周期和细胞凋亡。
结果
RNF138mRNA 在 5 种神经胶质瘤细胞系中均有表达，其在神经胶质瘤组织中的表达水平明显高于正常脑组织。下调 RNF138 表达可抑制 U251 细胞增殖，促进细胞凋亡，同时 S 期细胞减少，G2 期细胞增多。
结论
RNF138 基因在神经胶质瘤组织和神经胶质瘤细胞系中高表达，在神经胶质瘤细胞增殖、凋亡和细胞周期中发挥重要作用。
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