title: Down-regulation of STAT3 expression using vector-based RNA interference promotes apoptosis in Hepatocarcinoma cells.
abstract: In this study, we followed a DNA vector-based RNAi approach to silence the signal transducer and activator of transcription 3 (STAT3) expression in Bel-7402 cells, to explore how the Janus kinase (JAK)/STAT3 signaling pathway influences the apoptosis of hepatocarcinoma cells. According to GenBank's STAT3 cDNA, the plasmid pGCsi.U6/neoRFP STAT3, which was designed for expression of STAT3 small interfering RNA (siRNA), was constructed and synthesized, and then transfected into Bel-7402 cells using Lipofectamine 2000. Cells with or without siRNA transfection were treated in wells. The apoptotic rate was detected by flow cytometry (FCM) and by staining with the Annexin V/propidium iodide (PI) apoptosis detection kit. Simultaneously, the mitochondrial membrane potential (ΔΨm) was visualized by JC-1 fluorescence staining and observed using the inverted fluorescence microscope. Furthermore, the expression of caspase-3 protein was analyzed by Western blotting. The results showed that treatment with STAT3 siRNA displayed effects in the Bel-7402 cells, causing a significantly increased apoptotic ratio (P < 0.05). The mitochondrial membrane potential of the STAT3 siRNA group, observed by the JC-1 fluorescence staining, decreased significantly. The protein expression of active caspase-3 increased with STAT3 siRNA treatment, and was significantly higher than that of the control group (P < 0.05). STAT3 gene-silencing significantly improves the apoptotic effect against Bel-7402 cells.
authors.affiliation: a Institute of Oncology, Provincial Hospital Affiliated to Shandong University, Shandong University , Jinan , P.R. China.|b Department of Oncology , The Central Hospital of Panjin , Panjin , P.R. China.
authors.forename: Junwei
authors.initials: J
authors.lastname: Zhang
authors.affiliation: c Department of thoracic surgery , Provincial Hospital Affiliated to Shandong University, Shandong University , Jinan , P.R. China.
authors.forename: Jiajun
authors.initials: J
authors.lastname: Du
authors.affiliation: a Institute of Oncology, Provincial Hospital Affiliated to Shandong University, Shandong University , Jinan , P.R. China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Liu
authors.affiliation: d Department of General Surgery , The Central Hospital of Panjin , Panjin , P.R. China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics*; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism / Q000628:therapy; D005822:Genetic Vectors; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics; D034622:RNA Interference; D050796:STAT3 Transcription Factor / Q000096:biosynthesis / Q000235:genetics
chemical_list: D009363:Neoplasm Proteins; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human
keywords: Hepatocellular Carcinoma; RNAi; STAT3 gene; apoptosis