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Objective:To investigate the expression of livin and survivin induced by 5-Fu agent,and determine the role of livin and survivin in anti-chemotherapy of hepatocellular carcinoma(HCC). Methods:Low concentration of 5-Fu was added in cultured HepG2 cells.The expression of livin and survivin was detected by RT-PCR and western-blot on 24h and 48h after 5-Fu treatment.Results:Livin and survivin mRNA 1eve1 was increased by 0.6 ,1.1 and 1.5,2.2 times,respectively.and thant of the protein expression was increased by 1.3 ,2.7 and 0.9,1.9 times,respectively.Conclusion:livin and survivin was expression in HepG2.A high expression level of livin and survivin was induced in HepG2 by 5-Fu,which reduced to apoptosis by 5-Fu.It's valuable to study the curative effect of 5-Fu to hepatocelluar carcinoma further.
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Objective To investingate the effect of 5-Aza2'-deoxycytidine(5-Aza-CdR)on cell growth and to explore the possibility of re-expression of the hypermethylated and silenced RUNX3 gene in hepatocarcinoma cell line HepG2.Methods The change in expression of the tumor suppressor gene RUNX3 mRNA in cultured HepG2 cells was observed by RT-PCR before and after 5-Aza-CdR treatment.Activity of cell growth was observed by MTT assay and colony-forming test.The cell cycle was analyzed by flow cytometry.Apoptotic morphology was observed by transmitting electron microscopy.Results The gene was reactivated by two different doses of 5-Aza-CdR treatment in HepG2 cell without expressing RUNX3.The hepatocarcinoma cell line treated with 5-Aza-CdR displayed a slowed growth rate in contrast to the control group.The colony formation rate of HepG2 cell treated with 5-Aza-CdR decreased dramatically(P
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Objective To study the mechanism of killing and apoptosis-inducing effects of Capparis spinosa alkaloid (CSA) on human hepatocarcinoma cell line HepG2. Methods The killing effect of CSA on human hepatocarcinoma cell line HepG2 was studied by MTT method. Morphological observation of HepG2 cells was carried out by fluorescence microscope. Results The CSA had obvious cytotoxicity on the HepG2 in a dose-dependent manner and its IC50 value was 142.82 ?g/mL. The HepG2 cells showed the characteristic morphologic changes of apoptosis by the function of CSA and the apoptosis percentage is higher than that of the natural one. The progress of cells cycle from S phase to G2 phase had been blocked by CSA. The intracellular Ca2+ level had been increased by the function of CSA, which was positively related with drug concentration. Conclusion CSA has obviously killing and apoptosis-inducing effects on human hepatocarcinoma cell line HepG2 and calcium overload might also be invovled in these events.
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Objective: To further investigate the molecular mechanism of photodynamic therapy. Methods: We used cDNA microarray technique to explore the gene expression profiles of HEPG2 cells after photodynamic therapy with hematoperphyrin monomethyl ether(HMME) in HEPG2 cells. After treated with HMME for 60 min, the HEPG2 cells were irradiated with laser, and observed by microscope with H E staining. To prepare the probes, mRNA from both control and treated cells were isolated and purified, then reversely transcribed to cDNA with the incorporation of fluorecent labeled dUTP. The probes were hybridized with a cDNA microarray representing the 1 538 genes originated from human hepatocarcinoma cells. The fluorencent signals of Cy3 and Cy5 were scanned and analyzed. Results: After laser irradiation, the HEPG2 cells showed the typical feature of apoptosis. The gene expression profiles were also changed greatly. Among the 1 538 target genes, 389(2.47%) different expression genes were detected. Most of the changed genes (nearly 80%) were down regulated. They were functionally related to cell proliferation cycle, replication, metabolism and so on. Several apoptosis associated genes were detected among those up regulated genes, encoding the key proteins involved in apoptosis signal transduction, such as CCP32,AIF,Mch2. Conclusion: The HMME photodynamic therapy can initiate the apoptosis process of HEPG2 cells, which may be regulated by mitochondial pathway.[