›› 2019, Vol. 39 ›› Issue (12): 1375-.doi: 10.3969/j.issn.1674-8115.2019.12.006

• Original article (Basic research) • Previous Articles     Next Articles

Effects of connective tissue growth factor gene-silencing on cell growth and cell cycle of hepatic stellate cells

ZHANG Rong-hua, DONG An-ying, BAI Gan-ping, WAN Ping, JIANG Yi, WU Hong   

  1. 1. Department of Traditional Chinese Medicine and Rheumatology, Southwest Hospital, the Army Medical University of Chinese PLA, Chongqing 400038, China 2. Department of Heart, Kidney and Respiration, No.951 Hospital of PLA, Korla 841000, China
  • Online:2019-12-28 Published:2020-02-06
  • Supported by:
    National Natural Science Foundation of China, 81273918; Basic Research Military-Civil Integration Project, SWH2017JCZD-5; Specific Project of Traditional Chinese Medicine of Chinese PLA, 2010ZYZ231; Project of Traditional Chinese Medicine Science and Technology of Chongqing, 2012-2-63

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Abstract: Objective · To investigate the effects of silencing connective tissue growth factor (Ctgf) gene on the growth, cell cycle and the of TGF-β1, Smad3 and Smad7 of rat hepatic stellate cell line HSCT6. Methods · The recombinant lentivirus vector pCDH/Ctgf-shRNA of Ctgf gene was constructedRNA interference. The recombinant vector was packaged to obtain highly infectious pCDH/Ctgf-shRNA lentivirus particles for HSCT6 infection. The of green fluorescent protein (GFP) in the transfected HSCT6 cells was observed under fluorescence microscope. The effects of Ctgf-shRNA lentivirus on the growth of HSCT6 cells were testedCCK-8. The effects of Ctgf-shRNA lentivirus on the cell cycle of HSCT6 cells were analyzedflow cytometry (FCM). The effects of Ctgf-shRNA lentivirus on the of mRNA of Ctgf, Tgf-β1, Smad3 and Smad7, and their proteins in HSCT6 cells were detectedreal-time PCR and Western blotting, respectively. Results · The lentiviral vector pCDH/Ctgf-shRNA has been constructed successfully. The HSCT6 cells transfectedCtgf-shRNA lentivirus significantly expressed GFP under fluorescence microscope. The results of CCK-8 confirmed that the growth of HSCT6 cells transfectedCtgf-shRNA lentivirus was slower than that of controls and the differences were statistically significant after being cultured for 72 h (P0.05). The results of FCM revealed that the growth of HSCT6 cells transfectedCtgf-shRNA lentivirus was blocked in the S phase of cell cycle. The results of real-time PCR and Western blotting showed that the Ctgf-shRNA lentivirus effectively silenced Ctgf gene, down-regulated the of genes and encoding proteins of TGF-β1, and Smad3 of HSCT6 and up-regulated the of genes and encoding proteins of Smad7 of HSCT6 cells. The differences between transfected cells and controls were statistically significant (P0.05). Conclusion · Silencing Ctgf gene can effectively inhibit the growth of HSCT6 cells, down-regulate the of TGF-β1 and Smad3 and up-regulate the of Smad7. The inhibition of the growth of HSCT6 cells may be closely related to interference of the TGF-β1/Smads (Smad3 and Smad7) signaling pathway.

Key words: connective tissue growth factor (CTGF), hepatic stellate cell, transforming growth factor-β1 (TGF-β1), Smad3, Smad7