Construction and expression of STAT3 carrying EGFP and its inhibiting effect on HEP-2 cells

Objective To construct signal transduction and activators of transcription 3 ( STAT3) small interference RNA ( siRNA)expression vector and to detect the effect on inhibiting STAT3 expression. MethodsSTAT3 specific oligonucleotides were designed andsynthesized. These oligonucleotides were annealed to form the double strands DNA fragments.Then this fragment was cloned into pGPU6/EGFP/Neo vector. The recombinant STAT3-siRNA expressing construct was confirmed by using BbsⅠand BamHⅠdouble digestionand by sequencing. The STAT3-siRNA was transfected into HEP-2 cell. The inhibitory effect of STAT3-siRNA construct was examined with semi-quantitative reserve transcription PCR and Western blot. Cellular proliferation activities were assayed by tetrazolium bromide (MTT) colorimetry. ResultsSTAT3-siRNA expression vector was successfully constructed, and it could effectively reduce the mRNA and protein levels of STAT3 in transfected HEP-2 cells. HEP-2 cells transfected with STAT3-siRNA expression vector had lower cellular proliferation compared with no transfected HEP-2 cells (P0. 05). ConclusionThe STAT3-siRNA expression vector can effectively inhibit the expression of STAT3 and the proliferation of laryngeal cancer cells.