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Down-regulation of focal adhesion kinase by short hairpin RNA increased apoptosis of rat hepatic stellate cells

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Abstract:

An JY, Zheng LB, Xie SR, Dun ZN, Hao LS, Yao DM, Shih DQ, Zhang XL. Down-regulation of focal adhesion kinase by short hairpin RNA increased apoptosis of rat hepatic stellate cells. APMIS 2011; 119: 319–29.

Focal adhesion kinase (FAK) plays an essential role in the activation of hepatic stellate cells (HSC). The role of FAK on proliferation and apoptosis of fibronectin (FN)-stimulated HSC was investigated using short hairpin RNA (shRNA)-mediated gene silencing technology. FAK shRNA decreased the expressions of FAK, p-FAK (Tyr397), ERK1, and p-ERK1. FAK gene silencing also inhibited HSC proliferation by 11.08% at 12-h, 15.12% at 24-h, and 28.62% at 48-h post-transfection. Flow cytometric analysis (FACS) revealed that the apoptotic rate at 24 h was increased in the FAK shRNA plasmid group compared with the HK group (8.29 ± 0.79% vs 2.70 ± 0.31%, p < 0.01). TUNEL also confirmed the increase in the rate of apoptosis (19.00 ± 0.92% vs 7.63 ± 0.70%, p < 0.01), and studies showed that the caspase-3 expression was increased while the ratio of Bcl-2 to Bax was decreased. Together, these data show that FAK regulates HSC proliferation and induces the apoptosis of HSC via the caspase-3 and Bcl-2/Bax pathway.

Keywords: Apoptosis; RNA interference; focal adhesion kinase; hepatic stellate cells; proliferation

Document Type: Research Article

DOI: https://doi.org/10.1111/j.1600-0463.2011.02720.x

Affiliations: 1: Department of Gastroenterology, The Second Hospital of Hebei Medical University; Hebei Key Laboratory of Gastroenterology, Hebei Institute of Gastroenterology, No. 215 Heping West Road, 050000 Shijiazhuang, China 2: Inflammatory Bowel and Immunobiology Research Institute, Cedars-Sinai Medical Center, Los Angeles, California, USA

Publication date: 2011-06-01

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