上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

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1，闫国和2，汪国建1，粟永萍2   

  1. 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
  • 出版日期:2014-09-28 发布日期:2014-09-26
  • 作者简介: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。
  • 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-hua1, YAN Guo-he2, WANG Guo-jian1, SU Yong-ping2   

  1. 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:2014-09-28 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

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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。

关键词: 转化生长因子β1, 增殖细胞核抗原, 肝星状细胞-T6, 生长, 增殖

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.

Key words: transforming growth factor β1, proliferating cell nuclear antigen, hepatic stellate cell line T6, growth, proliferation