<em>ECHS1</em>经线粒体途径调控HepG2细胞凋亡

中国生物工程杂志

2013, Vol. 33

Issue (8): 11-16

研究报告

ECHS1经线粒体途径调控HepG2细胞凋亡

朱小三^1,2, 戴益琛¹, 陈章兴¹, 谢军培¹, 曾伟¹, 林园园¹, 赵本华³

1. 厦门大学附属成功医院厦门 361003;
2. 南昌大学研究生院医学部南昌 330006;
3. 厦门大学公共卫生学院厦门 361005

ECHS1 Involved in Antagonising Apoptosis in HepG2 Cells via the Mitochondrial Pathway

ZHU Xiao-san^1,2, DAI Yi-chen¹, CHEN Zhang-xing¹, XIE Jun-pei¹, ZENG Wei¹, LIN Yuan-yuan¹, ZHAO Ben-hua³

1. Department of Gastroenterology, Chenggong Hospital, Xiamen University, Xiamen 361003, China;
2. Medical department of Graduate College of Nanchang University, Nanchang 330006, China;
3. Department of Epidemiology and Health Statistics of Preventive Medicine, Xiamen University, Xiamen 361005, China

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摘要： 目的:观察烯脂酰辅酶A水合酶短链1(enoyl-CoA hydratase short chain 1, ECHS1)对HepG2细胞凋亡的影响。方法:构建 ECHS1 基因的干扰质粒,转染HepG2细胞后通过嘌呤霉素筛选稳定干扰 ECHS1 的细胞株(HepG2-si ECHS1),利用Western blot验证其干扰效率;利用流式细胞仪及TUNEL方法检测 ECHS1 干扰后HepG2细胞凋亡情况,再采用Western blot检测凋亡相关蛋白表达水平。结果:成功构建了 ECHS1 的干扰质粒;Western blot验证了 ECHS1 干扰后HepG2细胞株中 ECHS1 表达低于空白对照组(未转染HepG2细胞)及阴性对照组(空载体pU6转染HepG2细胞)(P<0.05);流式细胞仪术显示HepG2-si ECHS1 组细胞株凋亡率(14.98±1.07)%,阴性对照组(10.55±0.19)%,两者差异有统计学意义(P<0.05);TUNEL实验显示HepG2-si ECHS1 组细胞株凋亡率(6.13±0.12)%,阴性对照组(2.89±0.21)%,两者差异有统计学意义(P<0.05);与阴性对照组比较, ECHS1 干扰后p53、促凋亡蛋白Bax及Bid均表达增加。结论: ECHS1 通过线粒体途径拮抗HepG2细胞凋亡。

关键词： ECHS1; 线粒体途径; HepG2细胞; 凋亡

Abstract: Objective: To investigate the impact of enoyl-CoA hydratase short chain 1 ( ECHS1) on apoptosis of HepG2 cells. Methods: The ECHS1 interference plasmids were constructed and transfected into HepG2 cells. The stable interference cell lines were screened via puromycin. Interference efficacy was detected by Western blot. Flow cytometry and TUNEL assays studied the apoptosis of HepG2 cells. Apoptosis-related proteins were detected by Western blot. Results: The HepG2 cells with stable ECHS1 gene interference were successfully established. The expression level of ECHS1 protein in HepG2 cells after transfection with ECHS1 siRNA was significantly lower than that in the blank control cells (HepG2 cells without transfection) and the negative control cells (HepG2 cells transfected with pU6 vector) (P< 0.05). Apoptosis ratio of the ECHS1 siRNA group was significant higher than that of negative control group by both Flow cytometry and TUNEL assays (P<0.05). Expressions of p53, Bax and Bid in ECHS1 siRNA group were higher than those in negative control group. Conclusion: ECHS1 may antagonise the apoptosis of HepG2 cells via the mitochondrial pathway.

Key words: ECHS1 Mitochondrial pathway HepG2 cells Apoptosis

收稿日期: 2013-05-02 出版日期: 2013-08-25

ZTFLH:

Q819

基金资助: 南京军区医学科技创新课题(10MA070);厦门市科技计划医疗卫生创新项目(3502Z20124029)资助项目

通讯作者: 戴益琛,E-mail:dyichen@sina.com E-mail: dyichen@sina.com

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引用本文:

朱小三, 戴益琛, 陈章兴, 谢军培, 曾伟, 林园园, 赵本华. ECHS1经线粒体途径调控HepG2细胞凋亡[J]. 中国生物工程杂志, 2013, 33(8): 11-16.

ZHU Xiao-san, DAI Yi-chen, CHEN Zhang-xing, XIE Jun-pei, ZENG Wei, LIN Yuan-yuan, ZHAO Ben-hua. ECHS1 Involved in Antagonising Apoptosis in HepG2 Cells via the Mitochondrial Pathway. China Biotechnology, 2013, 33(8): 11-16.

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https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2013/V33/I8/11

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