天津医药

天津医药 ›› 2020, Vol. 48 ›› Issue (8): 700-704.doi: 10.11958/20193910

• 细胞与分子生物学 • 上一篇    下一篇

BNIP3介导的线粒体自噬对低氧环境下卵巢癌HO-8910PM细胞侵袭转移的影响#br#

陈国庆1，叶萍2，徐玲2，谢晓英2△   

  1. 1南方医科大学附属深圳妇幼保健院产科（邮编518028）；2赣南医学院第一附属医院妇产科
  • 收稿日期:2019-12-26 修回日期:2020-06-14 出版日期:2020-08-15 发布日期:2020-08-12
  • 通讯作者: 谢晓英 E-mail:xiexiaoying603@126.com
  • 作者简介:陈国庆（1982），男，博士，副主任医师，主要从事自噬与重大妇产科疾病的关系研究
  • 基金资助:
    国家自然科学基金资助项目（81360383）

Effects of mitophagy mediated by BNIP3 on invasion and metastasis of ovarian cancer HO-8910PM cells under hypoxia#br#

CHEN Guo-qing1, YE Ping2, XU Ling2, XIE Xiao-ying2△   

  1. 1 Department of Obstetrics, the Affiliated Shenzhen Maternity & Child Healthcare Hospital, Southern Medical University, Shenzhen 518028, China; 2 Department of Obstetrics and Gynecology, the First Affiliated Hospital of 
    Gannan Medical University
  • Received:2019-12-26 Revised:2020-06-14 Published:2020-08-15 Online:2020-08-12
  • Contact: XIE Xiao-ying E-mail:xiexiaoying603@126.com

PDF (PC)

6360

摘要： 目的　探讨常氧及低氧条件下卵巢癌HO-8910PM细胞线粒体外膜蛋白B淋巴细胞瘤-2基因/腺病毒E1B相互作用蛋白3（BNIP3）的表达变化及其对线粒体自噬和细胞侵袭转移的影响。方法　将人高转移性卵巢癌细胞株HO-8910PM分为常氧组、低氧组、低氧+BNIP3 siRNA组，各组细胞处理48 h后，应用吖啶橙染色及透射电镜观察细胞线粒体自噬的变化；划痕实验及Transwell小室检测细胞迁移和侵袭能力的改变；实时荧光定量PCR（qPCR）和Western blot分析BNIP3、自噬微管相关蛋白轻链3-Ⅱ（LC3-Ⅱ）的基因及蛋白表达变化。结果　在低氧条件下，吖啶橙染色结果显示HO-8910PM细胞线粒体自噬率显著增加，透射电镜结果显示HO-8910PM细胞线粒体自噬小体显著增多（P＜0.01）。转染BNIP3 siRNA后细胞线粒体自噬、细胞迁移率和侵袭率均显著受抑（P＜0.01）。低氧组BNIP3、LC3-Ⅱ的mRNA和蛋白表达明显增高，低氧条件下转染BNIP3 siRNA后BNIP3、LC3-Ⅱ的mRNA和蛋白表达明显下调（P＜0.01）。结论　在低氧条件下抑制卵巢癌HO-8910PM细胞BNIP3的表达可显著抑制线粒体自噬，降低细胞迁移和侵袭能力。

关键词: 线粒体, 自噬, 腺病毒E1B蛋白质类, 卵巢肿瘤, 肿瘤细胞, 培养的, 细胞运动, 肿瘤转移, 腺病毒E1B相互作用蛋白3

Abstract: Objective　To investigate the expression of B-cell lymphoma-2/Adenovirus E1B interacting protein 3 (BNIP3) of ovarian cancer HO-8910PM cells under normal and hypoxic conditions and its effects on mitophagy, cell invasion and metastasis. Methods　The human highly metastatic ovarian cancer cell line HO-8910PM was divided into three groups: normoxia group, hypoxia group and hypoxia+BNIP3 siRNA group. After 48 hours of treatment, mitophagy was detected by acridine orange staining and transmission electron microscopy. Migration and invasion of HO-8910PM cells were determined by scratch test and Transwell chamber assay. The mRNA and protein expressions of BNIP3 and LC3-Ⅱ were analyzed by qPCR and Western blot assay. Results　Under low oxygen conditions，acridine orange staining showed a significant increase in the incidence of mitophagy in HO-8910PM cells, and transmission electron microscopy showed a significant increase in the number of mitochondrial autophagy of HO-8910PM cells (P＜0.01).  After transfection of BNIP3 siRNA, mitochondrial autophagy, cell migration and invasion were significantly inhibited (P＜0.01). The mRNA and protein expressions of BNIP3 and LC3-Ⅱwere significantly increased in hypoxia group, which decreased significantly after transfection of BNIP3 siRNA under hypoxia (P＜0.01). Conclusion　The inhibition of BNIP3 expression in HO-8910PM cells under hypoxia can significantly inhibit mitophagy and reduce cell migration and invasion.

Key words: mitochondria, autophagy, adenovirus E1B proteins;ovarian neoplasms, tumor cells, cultured, cell movement, neoplasm metastasis, adenovirus E1B interacting protein 3

中图分类号: