Apoptosis in mtDNA-depleted cells and its transmitochondrial cybrids

李歆强; 凌贤龙; 周源; 何玉琦; 李诗伟; 晏斌

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Apoptosis in mtDNA-depleted cells and its transmitochondrial cybrids

更新时间：2026-01-09

- Apoptosis in mtDNA-depleted cells and its transmitochondrial cybrids
- Journal of Army Medical University Vol. 31, Issue 22, Pages: 2225-2227(2009)
- 作者机构：
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  CLC： R329.25;R394.3;R73-354
- Published：2009
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李歆强,凌贤龙,周源,何玉琦,李诗伟,晏斌.诱导细胞mtDNA缺失以及再转入线粒体后对肿瘤细胞凋亡的影响[J].陆军军医大学学报(原第三军医大学学报),2009,31(22):2225-2227.

　LI Xin-qiang,LING Xian-long,ZHOU Yuan,HE Yu-qi,LI Shi-wei,YAN Bin.Apoptosis in mtDNA-depleted cells and its transmitochondrial cybrids[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(22):2225-2227.
李歆强,凌贤龙,周源,何玉琦,李诗伟,晏斌.诱导细胞mtDNA缺失以及再转入线粒体后对肿瘤细胞凋亡的影响[J].陆军军医大学学报(原第三军医大学学报),2009,31(22):2225-2227. DOI：

　LI Xin-qiang,LING Xian-long,ZHOU Yuan,HE Yu-qi,LI Shi-wei,YAN Bin.Apoptosis in mtDNA-depleted cells and its transmitochondrial cybrids[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(22):2225-2227. DOI：

摘要

目的对比研究mtDNA缺失以及再转入线粒体后细胞凋亡的变化。方法在成功构建ρ

SK-Hep1细胞和转线粒体细胞SK-Hep1Cyb的基础上，采用Annexin V/PI 双染色法检测细胞凋亡；流式细胞仪检测细胞内活性氧（ROS）和线粒体膜电位（ΔΨm）；Western blot检测细胞Bcl-2、Bax表达水平；免疫荧光染色观测Bcl-2细胞内分布。结果 SK-Hep1、ρ

SK-Hep1和SK-Hep1Cyb细胞凋亡率分别为（2.01±0.11）%、（0.37±0.08）%和（2.10±0.12）％。ρ

SK-Hep1对细胞凋亡有明显抗性（P0.05）。ρ

SK-Hep1细胞内DCFDA荧光强度较 SK-Hep1细胞显著增强（35.5与15.9

P0.01）；转入线粒体后，SK-Hep1Cyb细胞DCFDA荧光强度较ρ

SK-Hep1细胞明显下降（17.4与35.5

P0.01）。ρ

SK-Hep1细胞MitoTracker Red荧光强度较SK-Hep1细胞显著减低（55.0与65.9

P0.05）；转入线粒体后，SK-Hep1Cyb细胞MitoTracker Red荧光强度与SK-Hep1细胞基本一致（67.4与65.9

P0.05）。ρ

SK-Hep1细胞线粒体Bcl-2、Bax表达增多，Bcl-2/Bax比值增加（P0.01）。SK-Hep1Cyb细胞线粒体Bcl-2/Bax值下降。结论 mtDNA缺失肿瘤细胞对细胞凋亡有明显拮抗。Bcl-2线粒体转位、线粒体Bcl-2/Bax值增加、ROS产生增多可能参与细胞凋亡拮抗的形成。

Abstract

Objective To investigate cell apoptosis in mtDNA-depleted SK-Hep1 cells and the transmitochondrial cybrids on the basis of establishment of ρ

SK-Hep1 (a mtDNA-depleted cell line derived from human hepatoma cell line SK-Hep1) and the transmitochondrial cybrids

SK-Hep1Cyb. Methods SK-Hep1 cells

SK-Hep1 cells

and the transmitochondrial cybrids were cultured in high-glucose DMEM. Apoptosis was determined by Annexin V/PI assay. Flow cytometry (FCM) and laser scan confocal microscopy were used to detect reactive oxygen species (ROS) and mitochondrial membrane potential (ΔΨm) respectively. Western blot analysis was applied to detect Bcl-2 and Bax expressions in the above 3 groups of cells. Distribution of Bcl-2 in the 3 groups was assessed by immunofluorescence analysis. Results Apoptotic ratio of SK-Hep1

SK-Hep1 and SK-Hep1Cyb cells were (2.01±0.11)%

(0.37±0.08)% and (2.10±0.12)％

respectively. ρ

SK-Hep1 cells were more counteractive against apoptosis than the other 2 cells (P0.05). Fluorescence intensity of ROS in ρ

SK-Hep1 cells was significantly higher than that of the parent cells (35.5 vs 15.6

P0.01)

but the ΔΨm was comparatively lower (55.0 vs 65.9

P0.05)

meanwhile the ratio of Bcl-2/Bax was elevated. After transmitochondria to ρ

SK-Hep1 cells

the SK-Hep1Cyb cells had all the above reversed. Conclusion mtDNA-depleted cells are more counteractive against apoptosis

and it might be related to the elevated mitochondrial Bcl-2/Bax and ROS.

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