[Knockdown of STAT3 inhibits proliferation and migration of HepG2 hepatoma cells induced by IFN1].

Objective To prepare lentiviruses expressing shRNA sequences targeting human signal transducer and activator of transcription 3 and detect the effect of knockdown on type I interferon (IFN1)-induced proliferation and migration in HepG2 cells. Methods Four shRNA sequences (shRNA1-shRNA4) and one control sequence (Ctrl shRNA) were selected and cloned respectively into pLKO.1-sp6-pgk-GFP to construct shRNA-expressing vectors. Along with backbone psPAX2 and pMD2.G vectors, they were separately transfected into HEK293T cells to prepare lentiviruses. HepG2 cells were infected with the lentiviruses. Cytoplastic duanyu18133 level was detected by Western blotting to screen effective shRNA sequence(s) targeting Proliferation and migration of HepG2 cells were analyzed by CCK-8 assay and Transwell^TM migration and scratching assay, respectively. To detect the effect of IFN1 on cell proliferation and migration of HepG2 cells, the cells were treated with 2000 U/mL IFNα2b for indicated time and the activation of IFN-triggered signal transduction was assayed by Western blotting. Results Two most effective duanyu18133-targeting shRNA sequences shRNA1 and shRNA2 were selected, and the expression of both duanyu18133 shRNA significantly decreased proliferation and migration of HepG2 cells. When treated with IFNα2b, 2000 U/mL of IFN1 showed more competent in attenuating growth and migration of HepG2 cells. Our data further proved that knockdown of duanyu18133 increased the phosphorylation of and IFNα2b further enhanced the activation of duanyu18131 signaling in HepG2 cells. Conclusion Knockdown of duanyu18133 inhibits cell migration and growth, and rescues IFN response through up-regulating duanyu18131 signal transduction in HepG2 hepatoma cells.

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