Effect of siRNA-mediated inhibition of nuclear transcription factor-kappa B on apoptosis of hepatocarcinoma cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effect of siRNA-mediated inhibition of NF-κB on apoptosis of hepatocarcinoma cells.</p><p><b>METHODS</b>Specific small interfering RNA Targeting NF-κB gene was synthesized and transfected into HepG2 cells by liposomes. Nested RT-PCR and quantitative RT-PCR were used to detect the mRNA expression of NF-κB. Immunohistochemistry, enzyme-linked immunosorbent assay and Western blot were performed to examine the protein expression of NF-κB. Annexin V-FITC was used to test cell apoptosis.</p><p><b>RESULTS</b>The expression of NF-κB in HepG2 cells (1.13+/-0.03) was significantly higher (t=27.02, P<0.05) than that in normal hepatocytes (0.29+/-0.07). The down-regulation of NF-κB expression was depended on the dosage of siRNA and the time after transfection. 72 h after siRNA transfection, NF-κB expression reduced by 93% and 62% at the mRNA and protein levels, respectively. The apoptosis of HepG2 cells increased by 85% with NF-κB inhibition.</p><p><b>CONCLUSIONS</b>NF-κB is abnormally active in HepG2 cells and NF-κB inhibition mediated by siRNA promotes HepG2 cells apoptosis. It suggested that NF-κB could be a potential target for HCC prevention gene therapy.</p>

Dynamic expression and quantitative analysis of hepatic nuclear factor-kappa B and its gene during the development of hepatocellular carcinoma / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the kinetic expression and alteration of nuclear transcription factor-kappa B (NF-kappaB) and its gene in hepatocellular carcinoma (HCC) development.</p><p><b>METHODS</b>A hepatoma model was established with N-(2-fluorenyl) acetamide (2-FAA) using male SD rats. Morphological changes and dynamic alterations of NF-kappaB and NF-kappaB mRNA of the rat livers at different stages of HCC development were observed by pathological examinations. The liver specimens from HCC patients were collected by self-control method. The expression of NF-kappaB was quantitatively analyzed by ELISA.</p><p><b>RESULTS</b>Hepatocytes showed vacuole-like denaturation, atypical hyperplasia, and transformation into highly differentiated cancerous hepatocytes with increasing tendencies of liver NF-kappaB and NF-kappaB mRNA expressions. The NF-kappaB positive material was granule-like and stained brown, with dot-nest-like staining localized in the nuclei and cytoplasm of HCC cells, but only in the cytoplasm of the cells of park cancer tissues. Its expression in HCC cells was stronger than that in their surrounding tissues (chi2 = 13.1, P less than 0.01). No positive relationship was found between NF-kappaB expression and histological grades, the number of tumors, or size of the tumors.</p><p><b>CONCLUSION</b>The expression of NF-kappaB and its gene are associated with the development of HCC. To inhibit the expression may be useful to HCC therapy.</p>