论著(基础研究)

沉默转化生长因子β1对肝星状细胞-T6生长及增殖的影响

  • 张荣华 ,
  • 闫国和 ,
  • 汪国建 ,
展开
  • 第三军医大学 1.西南医院中西医结合科, 2.军事预防医学院全军复合伤研究所创伤、烧伤与复合伤国家重点实验室, 重庆 400038
张荣华(1962—), 女, 教授, 主任医师, 博士, 硕士生导师; 电子信箱: zhrhlggg@163.com。

网络出版日期: 2014-09-26

基金资助

国家自然科学基金(81273918);重庆市中医药科技项目(2012-2-63);军队中医药科研专项课题(2010ZYZ231)

Effects of silencing TGFβ1 on growth and proliferation of hepatic stellate cell line T6

  • ZHANG Rong-hua ,
  • YAN Guo-he ,
  • WANG Guo-jian ,
  • et al
Expand
  • 1.Department of Integrated Chinese and Western Medicine, Southwest Hospital, the Third Military Medical University of Chinese PLA, Chongqing 400038, China; 2.State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Combined Injury, College of Military Preventive Medicine, the Third Military Medical University of Chinese PLA, Chongqing 400038, China

Online published: 2014-09-26

Supported by

National Natural Science Foundation of China,81273918;Project of Traditional Chinese Medicine Science and technology of Chongqing,2012-2-63;Specific Project of Traditional Chinese Medicine of Chinese PLA,2010ZYZ231

摘要

目的 观察沉默转化生长因子β1 (TGFβ1)对肝星状细胞(HSC)-T6生长及增殖的影响。方法 用具有高效感染力的TGFβ1 shRNA慢病毒液感染HSC-T6细胞,以未经感染或经空病毒感染的HSC-T6细胞作对照。倒置荧光显微镜下观察HSC-T6细胞绿色荧光蛋白(GFP)的表达变化;CCK-8法检测TGFβ1 shRNA慢病毒对HSC-T6细胞生长及增殖的影响;RT-PCR和Western blotting分别检测HSC-T6细胞TGFβ1及增殖细胞核抗原(PCNA)基因及其编码蛋白的表达,并予以定量分析。结果 经TGFβ1 shRNA慢病毒感染的HSC-T6细胞显著表达GFP。CCK-8法检测结果显示:经TGFβ1 shRNA慢病毒感染的HSC-T6细胞的生长及增殖慢于对照细胞,培养48 h后差异有统计学意义(P<0.01)。RT-PCR和Western blotting检测结果显示:TGFβ1 shRNA慢病毒能有效沉默HSC-T6细胞TGFβ1,并下调其核抗原PCNA基因及其编码蛋白的表达,与对照细胞的差异有统计学意义(P<0.01)。结论 沉默TGFβ1能明显抑制HSC-T6的生长及增殖,并下调其核抗原PCNA的表达。

本文引用格式

张荣华 , 闫国和 , 汪国建 , . 沉默转化生长因子β1对肝星状细胞-T6生长及增殖的影响[J]. 上海交通大学学报(医学版), 2014 , 34(9) : 1315 . DOI: 10.3969/j.issn.1674-8115.2014.09.012

Abstract

Objective To observe the effects of silencing transforming growth factor β1 (TGFβ1) on the growth and proliferation of hepatic stellate cell line T6 (HSC-T6). Methods The TGFβ1 shRNA lentiviral vector particles with high infectivity were transfected into HSC-T6 cells. The controls were HSC-T6 cells that were not infected or infected by empty virus. The expression of green fluorescent protein (GFP) of the HSC-T6 cells was observed under the inverted fluorescence microscope. The effects of TGFβ1 shRNA lentivirus on the growth and proliferation of HSC-T6 cells were detected by the CCK-8. The expressions of the gene and encoding protein of TGFβ1 and proliferating cell nuclear antigen (PCNA) of HSC-T6 cells were detected by the RT-PCR and Western blotting, respectively. The quantitative analysis was then conducted. Results HSC-T6 cells transfected by TGFβ1 shRNA lentivirus significantly expressed GFP. The results of CCK-8 showed that the growth and proliferation of HSC-T6 cells transfected by TGFβ1 shRNA lentivirus were slower than those of controls and the differences were statistically significant after being cultured for 48 h (P<0.01). The results of RT-PCR and Western blotting revealed that the TGFβ1 shRNA lentivirus effectively silenced the TGFβ1 and down-regulated the expressions of gene and encoding protein of PCNA of HSC-T6 cells. The differences of transfected cells and controls were statistically significant (P<0.01). Conclusion Silencing TGFβ1 can effectively inhibit the growth and proliferation of HSC-T6 cells and down-regulate the expression of PCNA.

文章导航

/