Effect of Extract on Cell Proliferation and Apoptosis of U251 Cells and Its Molecular Mechanism

GU Hao; WANG Hai-bo; FENG Jun; QIAN Ya-yun; JIN Feng; CHEN Jue; SHI You-yang; LU Song-hua; XU Pei; LIU Yan-qing

doi:10.13422/j.cnki.syfjx.2017170115

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Effect of Extract on Cell Proliferation and Apoptosis of U251 Cells and Its Molecular Mechanism

更新时间：2024-01-04

- Effect of Extract on Cell Proliferation and Apoptosis of U251 Cells and Its Molecular Mechanism
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 23, Issue 17, Pages: 115-120(2017)
- 作者机构：
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.2017170115
  CLC： R285
- Published：2017
- 稿件说明：
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GU Hao, WANG Hai-bo, FENG Jun, et al. Effect of Extract on Cell Proliferation and Apoptosis of U251 Cells and Its Molecular Mechanism[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(17): 115-120.
DOI：

GU Hao, WANG Hai-bo, FENG Jun, et al. Effect of Extract on Cell Proliferation and Apoptosis of U251 Cells and Its Molecular Mechanism[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(17): 115-120. DOI： 10.13422/j.cnki.syfjx.2017170115.

摘要

目的：探讨南蛇藤茎乙酸乙酯提取物（Celastrus orbiculatus extract，COE）抗人胶质母细胞瘤U251细胞株的增殖和凋亡作用，研究COE抗肿瘤的分子机制，为寻找新的治疗胶质母细胞瘤药物提供依据。方法：以人胶质母细胞瘤细胞株U251为研究对象，设空白组和COE组，应用噻唑蓝[3-（4，5-dimethyl-2-thiazolyl）-2，5-diphenyl-2-H-tetrazolium bromide，MTT]比色法观察COE对细胞活力的抑制作用；5-溴脱氧尿嘧啶核苷（5-bromo-2-deoxy uridine

BrdU）标记检测COE对U251细胞增殖的影响；Annexin V/碘化丙啶（PI）双标法检测COE对U251细胞凋亡的作用；透射电镜下观察经COE干预后U251细胞超微结构变化；蛋白免疫印迹法（Western blot）检测COE对凋亡标志基因B淋巴细胞瘤-2（B-cell lymphoma-2，Bcl-2）

Bcl-2相关X蛋白（Bcl-2-associated X，Bax）和半胱氨酸蛋白酶-3（Caspase-3）蛋白表达的影响。结果：与空白组比较，COE能够明显抑制U251细胞的增殖（P<0.05）；诱导U251细胞的早期凋亡，电镜下可见到凋亡小体，影响并改变U251细胞超微结构；COE能促进Caspase-3，Bax蛋白表达，降低Bcl-2，Bcl-2/Bax蛋白表达（P<0.05）。结论：COE能有效抑制胶质母细胞瘤U251细胞株的增殖，促进肿瘤细胞凋亡，其作用机制与调节Bcl-2和Caspase-3表达有关。

Abstract

Objective: To observe the effect of different concentrations of Celastrus orbiculatus extract (COE) on proliferation and apoptosis of U251 cells (a glioblastoma cancer cell line)

and its possible molecular mechanism. Method: U251 cells at the logarithmic growth phase were divided into control group and COE-treated group. 3-(4

5-dimethyl-2-thiazolyl)-2

5-diphenyl-2-H-tetrazolium bromide(MTT) assay and 5-bromo-2-deoxy uridine(BrdU) incorporation experiment were performed to observe the inhibitory effect on cell viability and proliferation

respectively. Annexin V/PI double staining was used to detect the effect of COE on U251 cell apoptosis. Ultrastructural changes of U251 cells after COE treatment were observed under transmission electron microscope; the protein expressions of apoptosis marker B-cell lymphoma-2/Bcl-2-associated X (Bcl-2/Bax) and Caspase-3 were detected by Western blot. Result: Compared to control group

the proliferation of U251 cells in COE-treated group were significant inhibited by COE (P<0.05). COE could induce the early apoptosis and ultrastructure changes of U251 cells

and apoptotic bodies were observed under transmission electron microscope. The protein expressions of Bcl-2/Bax and Caspase-3 were changed in a concentration dependent manner. Conclusion: COE can effectively inhibit the proliferation of U251 cells by down-regulating the expression of Bcl-2/Bax and increasing the protein expression of Caspase-3

and promoting the apoptosis of cell

suggesting that COE can be used as a drug candidate for treating glioblastoma.

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