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Rab3b 在人脑胶质瘤中的表达:对细胞增殖和凋亡的影响。

Expression of Rab3b in Human Glioma: Influence on Cell Proliferation and Apoptosis.

机构信息

The First School of Clinical Medicine, Southern Medical University, Guangzhou City, Guangdong 510515, China.

Department of Medical Laboratory, General Hospital of the Central Theater Command of the Chinese People's Liberation Army, Wuhan City, Hubei Province, 430070, China.

出版信息

Curr Pharm Des. 2021;27(7):989-995. doi: 10.2174/1381612826666200917145228.

Abstract

BACKGROUND

Glioma is the most common human central nervous system tumour with a high degree of malignancy. Some Rab GTPases have significant effects on glioma.

OBJECTIVE

This study aimed to investigate the effect of Rab3b (Rab GTPase3b) on human glioma cell proliferation and apoptosis by silencing Rab3b and to initially verify the value of Rab3b expression for the diagnosis and progression in human glioma.

METHODS

Rab3b was silenced by siRNA transfection. Human glioma tissues and normal brain tissues adjacent to glioma were obtained by surgery. Rab3b, P53, Caspase 7, Bax, and Bim mRNA and protein expression levels were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. Cell proliferation was detected by the cell counting kit-8 assay, and the cell cycle and apoptosis were analysed using flow cytometry.

RESULTS

Rab3b mRNA and protein expression in human glioma U251 and U87 cells were significantly downregulated after Rab3b silencing. Rab3b silencing inhibited glioma cell proliferation by promoting cell cycle arrest and induced apoptosis by upregulating the expression of apoptosis-related proteins. Rab3b expression in human glioma (n = 33) was significantly higher than that in normal brain tissues adjacent to glioma (n = 15). In addition, Rab3b expression levels in high-grade gliomas (WHO III-IV, n = 19) were also significantly higher than those in low-grade gliomas (WHO I-II, n = 14).

CONCLUSION

Rab3b expression levels are significantly related to the progression of gliomas. Moreover, Rab3b silencing not only significantly inhibits cell proliferation in gliomas via cell cycle arrest but also promotes cell apoptosis by upregulating the expression levels of apoptosis-related proteins; however these preliminary in vitro results warrant validation on in vivo studies.

摘要

背景

神经胶质瘤是最常见的人类中枢神经系统肿瘤,具有高度恶性。一些 Rab GTPases 对神经胶质瘤有重要影响。

目的

本研究旨在通过沉默 Rab3b 来研究 Rab3b(Rab GTPase3b)对人神经胶质瘤细胞增殖和凋亡的影响,并初步验证 Rab3b 表达对人神经胶质瘤的诊断和进展的价值。

方法

通过 siRNA 转染沉默 Rab3b。通过手术获得人神经胶质瘤组织和神经胶质瘤旁正常脑组织。通过实时定量聚合酶链反应(qRT-PCR)和 Western blot 检测 Rab3b、P53、Caspase 7、Bax 和 Bim mRNA 和蛋白表达水平。通过细胞计数试剂盒-8 检测细胞增殖,通过流式细胞术分析细胞周期和凋亡。

结果

沉默 Rab3b 后,人神经胶质瘤 U251 和 U87 细胞中 Rab3b mRNA 和蛋白表达水平明显下调。Rab3b 沉默通过促进细胞周期停滞抑制神经胶质瘤细胞增殖,并通过上调凋亡相关蛋白的表达诱导细胞凋亡。Rab3b 在人神经胶质瘤(n = 33)中的表达明显高于神经胶质瘤旁正常脑组织(n = 15)。此外,高级别神经胶质瘤(WHO III-IV,n = 19)中 Rab3b 的表达水平也明显高于低级别神经胶质瘤(WHO I-II,n = 14)。

结论

Rab3b 表达水平与胶质瘤的进展明显相关。此外,Rab3b 沉默不仅通过细胞周期停滞显著抑制神经胶质瘤细胞增殖,而且通过上调凋亡相关蛋白的表达水平促进细胞凋亡;然而,这些初步的体外结果需要在体内研究中进行验证。

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