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Effect of siRNA-Mediated Inhibition of Nuclear Transcription Factor-κB on Apoptosis of Hepatocellular Carcinoma Cells

Author: WangYiLang
Tutor: YaoDengFu
School: Nantong University
Course: Oncology
Keywords: Hepatocellular carcinoma Hepatitis B virus Nuclear factor-κB Small interfering RNA Sequencing Apoptosis
CLC: R735.7
Type: Master's thesis
Year: 2009
Downloads: 50
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Abstract


Objective: Hepatocellular carcinoma (HCC) is one of the most common malignant tumor , the ongoing hepatitis virus infection caused by chronic inflammation of the liver is the major morbidity background . Nuclear transcription factor -κB (NF-κB) overexpression in tumor cells often inhibit apoptosis induced proliferation and promote tumor formation . Studies have shown that abnormal activation of NF-κB in HCC often , but and the relationship between HBV replication and its formation in HCC apoptosis inhibitory mechanism is unclear . This study investigated the relationship of NF-κB expression in HBV-related HCC tissue characteristics and clinical pathology , and small interfering RNA (siRNA) to inhibit NF-κB signaling pathway, analysis of its impact on HepG2 cell apoptosis . Methods: Using immunohistochemistry method were detected 35 cases of HCC cancer tissue and its control of cancer adjacent tissues NF-κB expression ; biotin-labeled HBV - DNA probe to detect cancerous tissue HBV -DNA , and analyze NF-κB in the relationship between HBV replication its clinicopathological features . NF-κB expression in HepG2 cells with siRNA ; using the polymerase chain reaction , immunohistochemistry method , enzyme-linked immunosorbent assay and Western blot detection of NF-κB expression level ; HepG2 cell apoptosis using Annexin V-FITC detection , analysis The relationship between inhibition of NF-κB expression and apoptosis in HepG2 cells . Results: NF-κB in cancer tissue was spotty expression , located in the cytoplasm and nucleus ; adjacent tissues NF-κB no positive expression mainly in the cytoplasm and nucleus . Cancer of NF-κB expression intensity was significantly higher than the adjacent tissues ; cancer tissue NF-κB expression was 100% cancer tissue was 68.6% , the difference was significant (Fisher's value = 0.000 ) . NF-κB expression intensity and degree of tumor differentiation , number of tumors and tumor diameter was no correlation between clinical and pathological features of cancer tissue . NF-κB in HBV-DNA - positive HCC tissue than concentration was significantly higher than the HBV - DNA negative group ( t = 4.7347 , P = 0.000 ) . siRNA transfected HepG2 cells after 72 h , NF-κB mRNA and protein expression were decreased by 93% and 62 % , siRNA down NF-κB in a dose and time dependent inhibition of NF-κB in a 85% increase in apoptosis in HepG2 cells . Conclusion : NF-κB activation abnormalities occur with HBV-related HCC development are closely related , siRNA effectively inhibited HepG2 cells NF-κB expression and induce apoptosis , NF-κB HCC gene therapy is expected to become an ideal target .

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Liver tumors
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