[1]吴亚冉,倪振洪,闫小晶,等.盐酸吡柔比星通过上调ATG4B增强HeLa细胞保护性自噬[J].第三军医大学学报,2016,38(02):129-135.
 Wu Yaran,Ni Zhenhong,Yan Xiaojing,et al.Pirarubicin induces cytoprotective autophagy in HeLa cells through up-regulating autophapy related 4B[J].J Third Mil Med Univ,2016,38(02):129-135.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
38卷
期数:
2016年第02期
页码:
129-135
栏目:
基础医学
出版日期:
2016-01-30

文章信息/Info

Title:
Pirarubicin induces cytoprotective autophagy in HeLa cells through up-regulating autophapy related 4B
作者:
吴亚冉倪振洪闫小晶郑英如何凤田连继勤
第三军医大学基础医学部生物化学与分子生物学教研室; 第三军医大学大坪医院野战外科研究所妇产科
Author(s):
Wu Yaran Ni Zhenhong Yan Xiaojing Zheng Yingru He Fengtian Lian Jiqin

Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Third Military Medical University, Chongqing, 400038; Department of Obstetrics and Gynecology, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing, 400042, China

关键词:
盐酸吡柔比星自噬自噬相关蛋白4B宫颈癌
Keywords:
pirarubicin autophagy autophapy related 4B uterine cervical neoplasm
分类号:
R737.33;R966;R979.1
文献标志码:
A
摘要:

目的       探讨盐酸吡柔比星增强宫颈癌HeLa细胞自噬水平的可能机制。       方法       采用CCK-8检测盐酸吡柔比星单独或联用自噬抑制剂氯喹对HeLa细胞增殖的影响;台盼蓝染色实验检测盐酸吡柔比星单独或联用自噬抑制剂氯喹对HeLa细胞死亡的影响;GFP-LC3 质粒转染HeLa细胞后观察盐酸吡柔比星对自噬小体形成的影响;Western blot检测细胞中自噬标志分子Ⅱ型微管相关蛋白轻链3(microtuble-associated protein light chain 3-Ⅱ, LC3-Ⅱ)、自噬相关分子自噬相关蛋白4B(autophapy related 4B,ATG4B)的表达情况。CCK-8实验及台盼蓝染色实验观察在有或者无自噬抑制剂氯喹作用下以及shRNA下调HeLa细胞中的ATG4B后盐酸吡柔比星对HeLa细胞的杀伤效应的影响。       结果       盐酸吡柔比星能剂量依赖地抑制HeLa细胞的增殖并促进其死亡(P<0.05);盐酸吡柔比星处理能显著增加HeLa 细胞中自噬小体的数量,并显著上调自噬标志分子LC3-II、自噬关键酶ATG4B的表达;自噬抑制剂氯喹可显著增强盐酸吡柔比星的杀细胞效应(P<0.05),shRNA干扰ATG4B可抑制盐酸吡柔比星对HeLa细胞中自噬的增强作用(P<0.05),并增强盐酸吡柔比星的杀细胞效应(P<0.05)。       结论       盐酸吡柔比星通过上调ATG4B而增强的自噬对HeLa细胞起保护作用。

Abstract:

Objective       To investigate the mechanism and significance of pirarubicin in enhancing the autophagy of HeLa cells.        Methods       Trypan blue exclusion assay and CCK-8 assay were used to detect cell mortality and proliferation in HeLa cells treated with different doses of pirarubicin alone or combined with chloroquine (autophagy inhibitor). The effect of pirarubicin on autophagosome formation in HeLa cells transfected with GFP-LC3 plasmid was observed with fluorescence microscopy. Western blotting was used to determine the protein levels of autophagy markers microtuble-associated protein light chain 3-Ⅱ (LC3-Ⅱ), and autophapy related 4B (ATG4B). After ATG4B was down-regulated with shRNA in HeLa cells, the effect of pirarubicin on killing HeLa cells in the presence of chloroquine was observed.        Results       Pirarubicin inhibited cell proliferation and induced cell death in a dose-dependent manner in HeLa cells (P<0.05). Pirarubicin induced the formation of LC3-GFP puncta in HeLa cells, and increased the expression of LC3Ⅱand ATG4B markedly. Chloroquine enhanced pirarubicin-triggered cell death in HeLa cells (P<0.05). Knockdown of ATG4B via shRNA suppressed pirarubicin-induced autophagy and increased pirarubicin-induced cell death in HeLa cells.        Conclusion       Pirarubicin induces cytoprotective autophagy in HeLa cells through up-regulating ATG4B.

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更新日期/Last Update: 2016-01-14