微信公众号

官网二维码
中国癌症防治杂志

中国癌症防治杂志 ›› 2023, Vol. 15 ›› Issue (2): 129-137.doi: 10.3969/j.issn.1674-5671.2023.02.03

• 基础研究 • 上一篇    下一篇

BLOC⁃3介导Rab32线粒体定位改变对肝癌细胞生长的作用研究

刘小丽,廖达文,王星琛,许小君,魏元元,王楠,张志刚,孙夏承,黄启超,季乐乐   

  1. 空军军医大学基础医学院生理与病理生理学教研室；陕西中医药大学第二临床医学院；延安大学基础医学院生理与病理生理学教研室；空军军医大学基础医学院教学实验中心
  • 出版日期:2023-04-25 发布日期:2023-05-05
  • 通讯作者: 季乐乐 E-mail：jilele@fmmu.edu.cn； 黄启超 E-mail：huangqichao1@163.com
  • 基金资助:
    国家自然科学基金青年科学基金项目（82002518）

Effect of the BLOC 3 mediated mitochondrial localization of Rab32 on the  growth of liver cancer cells

  • Online:2023-04-25 Published:2023-05-05

PDF (PC)

260

赞

摘要： 目的 观察溶酶体相关细胞器生物发生复合体⁃3 （biogenesis of lysosome⁃related organelles complex⁃3，BLOC⁃3）亚基Hps1和Hps4对肝癌细胞中Rab32线粒体定位的影响及在肝癌细胞生长中的作用。 方法 通过公共数据库GenDoma，String和InBio Discover分析Hps1和Hps4与Rab32的相互作用情况。体外培养人肝癌细胞系SNU⁃739和Hep⁃3B，利用脂质体转染方法分别转染Hps1和Hps4相关的siRNAs和质粒，采用免疫荧光和Western blot观察Hps1和Hps4对Rab32线粒体定位的影响，细胞划痕、克隆形成、EdU、MTS及Transwell侵袭实验检测Hps1和Hps4调控Rab32线粒体定位后肝癌细胞迁移、增殖和侵袭的变化情况。通过公共数据库UALCAN中的数据集CPTAC分析Rab32蛋白在肝癌和正常肝脏组织中的表达差异。 结果 数据库分析结果显示，Hps1和Hps4均可以与Rab32相互作用；与正常肝脏组织相比，Rab32蛋白在肝癌组织中的表达显著降低（P<0.001）。在肝癌细胞系SNU⁃739中干涉Hps1或Hps4或同时干涉Hps1和Hps4后，Rab32线粒体定位均减少（均P<0.01），细胞线粒体Rab32蛋白表达均降低（均P<0.001），细胞增殖、迁移和侵袭能力增强（均P<0.05）；在肝癌细胞系Hep⁃3B中过表达Hps1和Hps4后，Rab32线粒体定位增多（P<0.01），细胞线粒体Rab32蛋白表达增加（P<0.001），细胞增殖、迁移和侵袭能力均受抑制（均P<0.01），而单独过表达Hps1或Hps4时无显著抑制作用（均P>0.05）。结论 BLOC⁃3亚基Hps1和Hps4均可与Rab32相互作用并增加Rab32线粒体定位，进而抑制肝癌细胞生长。

关键词: 肝癌, BLOC-3, Rab32, 线粒体, 迁移, 侵袭

Abstract: Objective To observe the effect of the Hps1 and Hps4 subunits of biogenesis of lysosome⁃related organelles complex⁃3(BLOC⁃3) on the mitochondrial localization of Rab32 in liver cancer cells and the role of  liver cancer growth. Methods The interaction of Hps1 and Hps4 with Rab32 was analyzed by public databases GenDoma, String and InBio Discover. Human liver cancer cell lines SNU⁃739 and Hep⁃3B were cultured in vitro and transfected with Hps1 and Hps4⁃related siRNAs and plasmids, respectively, by using the liposome transfection method. The effect of Hps1 and Hps4 on the mitochondrial localization of Rab32 was observed by immunofluorescence and Western blot. Cell scratch, clonogenesis, EdU, MTS and Transwell invasion assays were used to detect the changes of liver cancer cell migration, proliferation and invasion after the regulation of Rab32 mitochondrial localization by Hps1 and Hps4. The expression difference of Rab32 protein in liver cancer and normal liver tissues was analyzed by using dataset CPTAC in the public database UALCAN. Results Database analysis showed that both Hps1 and Hps4 could interact with Rab32. Rab32 protein expression was significantly decreased in liver cancer tissues compared with normal liver tissues (P<0.001). After interference with Hps1 or Hps4 or both Hps1 and Hps4 in liver cancer cell line SNU⁃739, the mitochondrial localization of Rab32 were decreased (all P<0.01), mitochondrial Rab32 protein expression were decreased (allP<0.001), and cell proliferation, migration and invasion were enhanced (all P<0.05). After overexpression of Hps1 and Hps4 in liver cancer cell line Hep⁃3B, the mitochondrial localization of Rab32 was increased (P<0.01), the protein expression of mitochondrial Rab32 was increased (P<0.001), and the proliferation, migration and invasion of cells were inhibited (all P<0.01), while the overexpression of Hps1 or Hps4 alone had no significant inhibitory effect (all P>0.05).  Conclusions Both Hps1 and Hps4, the subunits of BLOC⁃3, can interact with Rab32 and increase the mitochondrial localization of Rab32, thereby inhibiting the growth of liver cancer  cells.

Key words: Liver cancer , BLOC-3, Rab32, Mitochondria, Migration, Invasion

中图分类号: 

  • R735.7