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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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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