肝胆胰外科杂志 ›› 2019, Vol. 31 ›› Issue (6): 360-365.doi: 10.11952/j.issn.1007-1954.2019.06.010

• 论著 基础研究 • 上一篇    下一篇

外源性CC10基因表达对肝癌细胞增殖、 凋亡、迁移和侵袭的影响

张怀波,马荣龙,张德景    

  1. 河南省濮阳市油田总医院   普外三科,河南   濮阳   457001

  • 收稿日期:2018-10-22 出版日期:2019-06-27 发布日期:2019-07-28
  • 作者简介:张怀波(1976-),男,山东阳谷人,副主任医师,硕士。

Effects of exogenous CC10 gene expression on proliferation, apoptosis, migration and invasion of hepatoma carcinoma cells

ZHANG Huai-bo, MA Rong-long, ZHANG De-jing.   

  1. Department of General Surgery, Puyang Oilfield General Hospital, Puyang, Henan 457001, China 
  • Received:2018-10-22 Online:2019-06-27 Published:2019-07-28

摘要: 目的 探究外源性Clara细胞分泌蛋白10(CC10)的基因表达对肝癌细胞增殖、凋亡、迁移和 侵袭的影响,并探讨其可能的作用机制。方法 采用RT-qPCR和Western blotting检测人正常肝细胞LO2和 肝癌细胞HepG2、Hep3B中CC10的内源表达水平;将HepG2细胞系分为pcDNA 3.1组和pcDNA 3.1-CC10 组,用LipofectionTM分别转染pcDNA 3.1空载体质粒和重组质粒pcDNA 3.1-CC10。用CCK-8法、TUNEL 法和Transwell小室实验分别检测两组细胞增殖、凋亡情况及迁移、侵袭能力。采用RT-qPCR和Western blotting检测两组细胞中CC10和细胞周期蛋白D1(Cyclin D1)的表达水平。结果 与正常肝细胞LO2相 比,CC10在肝癌细胞HepG2、Hep3B中呈明显低表达(P<0.05);转染 pcDNA 3.1-CC10后的HepG2细胞内 CC10 mRNA和蛋白水平均显著增高(P<0.05)。与 pcDNA 3.1组相比,pcDNA 3.1-CC10组细胞活力受到抑 制,细胞活力以时间依赖性方式降低(P<0.05),细胞迁移和侵袭实验中的过膜细胞数均低于pcDNA 3.1组 (P<0.05),细胞凋亡率高于pcDNA 3.1组(P<0.05)。同时,Cyclin D1的mRNA和蛋白表达水平均明显降低 (P<0.05)。 结论? 外源性CC10基因表达可抑制肝癌细胞增殖、迁移和侵袭,促进其凋亡,其机制可能与 下调Cyclin D1表达有关。 

关键词: Clara细胞分泌蛋白10, 癌, 肝细胞, 细胞周期素D1, 细胞凋亡 

Abstract:

objective  To investigate the influence and the possible molecular mechanism of exogenous Clara cells protein 10 (CC10) gene expression on proliferation, apoptosis, migration and invasion of hepatoma carcinoma cells. Methods The endogenous expressions of CC10 in the LO2, HepG2 and Hep3B cells were detected by RT-qPCR and Western blotting. The HepG2 cell line was divided into pcDNA 3.1 group and pcDNA 3.1-CC10 group, which was transfected with pcDNA 3.1 and pcDNA 3.1-CC10 respectively. The CCK-8 assay, TUNEL staining and Transwell chamber experiment were respectively employed to examine the biological functions of CC10 on proliferation, apoptosis, and migration and invasion of the HepG2 cells. The mRNA and protein levels of CC10 and cell cycle protein D1 (Cyclin D1) were determined by RT-PCR and Western blotting. Results The CC10 expression was notably lower-expressed in both HepG2 cells and Hep3B cells as compared with LO2 cells (P<0.05), the mRNA and protein levels were significantly increased by transfection with pcDNA 3.1-CC10 (P<0.05). Compared with the pcDNA 3.1 group, cell viability of pcDNA 3.1-CC10 group was inhibited and decreased in time-dependent manner (P<0.05); the number of transmembrane cells was less than that in the pcDNA 3.1 group (P<0.05); apoptosis rate of pcDNA 3.1-CC10 group was obviously increased (P<0.05). The mRNA and protein levels of Cyclin D1 in the pcDNA 3.1-CC10 group decreased significantly (P<0.05). Conclusion Exogenous CC10 gene expression can inhibit the proliferation, migration and invasion, promote apoptosis of hepatoma carcinoma cells, and its mechanism may be related to the down-regulation of Cyclin D1 expression. 

Key words: Clare cell protein 10 (CC10), hepatocellualr carcinoma, cell cycle protein D1 (Cyclin D1), cell apoptosis

中图分类号: 

  • R735.7  
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