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The effects of DHAA-urea,a novel dehydroabietylamine(DHAA) derivatives,on cell viability and glucose metabolism,in hypoxia and normoxia human hepatoma HepG2 cells were investigated.Hypoxia cells were achieved using DMEM containing high concentration of glucose without serum and pre-incubating of CoCl_2 (final concentration 150 μmol/L) for 24 h.The antiproliferation effect of DHAA-urea was measured by colorimetric MTT assay.The cellular ATP concentration,the lactate dehydrogenase(LDH) and glucose-6-phosphate dehydro genase (G6PD) activity were detected by their kits.It was shown that DHAA-urea markedly inhibited cell viability,cellular ATP level,LDH and G6PD activity in either aerobic or anaerobic circumstance in a dose-and time dependent manner.This suggested that DHAA-urea possibly inhibited HepG2 cells growth via the inhibition of glucolysis and glucolysis-dependent ATP depletion.DHAA-urea could be a promising candidate in the development of a novel class of agents used for human hepatocellular carcinoma.
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Objective To assess the effects of p27mt gene transfection on the proliferation and apoptosis of human hepatocellular carcinoma cell (HCC) lines SMMC-7721. Methods A replication deficient adenovirus vector encoding p27mt (Ad-p27mt) was used and p27mt cDNA was transfected into human SMMC-7721 cell lines in vitro. The synthesis of DNA in SMMC-7721 cells was determined by using 3H-thymidine incorporation; the cell apoptosis was determined by flow cytometry, TUNEL method and DNA fragmentation analysis. Results The virus titer was 7.95?1012 cfu/ml, the transduction efficiency was 100 % when multiplicity of infection ≥50, FCM analysis revealed a sub-G1 cell peak in Ad-p27mt transduced hepatocellular carcinoma cell lines. Agarose electrophoresis showed marked ladder .The difference of apoptotic index between the Ad-p27mt group and the control group was statistically significant (58.6?4.3, vs 4.5?1.6, P
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OBJECTIVE:To investigate the serumpharmacological effects of Compound Banmao Capsule(FBC)on prolif?eration of SMMC-7721cells in human hepatocellular carcinoma.METHODS:The effects of FBC were investigated in vitro using serum pharmacological approach.Different doses(clinic equivalent dosage,duplation and triplicity)of FBC suspension were irrigated into the rabbit stomachs.The inhibitory effects of FBC serum on SMMC-7721cells was observed by MTT assay.The inhibitory effects at different collecting hours in the clinical equivalent group were also observed.RESULTS:All of the drug serums in the three groups could inhibit the proliferation of SMMC-7721cells(P
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AIM To elucidate whether apoptosis is involved in the development of multidrug resistance in 5-fluorouracil resistant cells Bel7402/5-FU. METHODS Multidrug resistance of Bel7402/5-FU cells was determined by SRB assay, apoptosis was measured by Annexin V-FITC/PI double labeled assay, cell cycle phases were analysed by flow cytometry, and expression of genes related to apoptosis bcl-2, bcl-xl, bcl-xs, bax, fas, p53, and cpp32 was detected by RT-PCR assay. RESULTS Bel7402/5-FU cells were resistant not only to 5-FU but also to multiple anticancer drugs. When treated with the same concentration of 5-FU (30 ?mol?L -1 or 150 ?mol?L -1 ), the proportion of apoptosis was significantly increased and G_0/G_1 phase was increased and S and G_2/M phases were reduced in Bel7402 cells, but the proportion of apoptosis and cell cycle was not changed in Bel7402/5-FU cells. The expression of bcl-xl, bcl-xs, and bax mRNA were significantly increased and the expression of p53 and cpp32 mRNA were significantly decreased in resistant Bel7402/5-FU cells, as compared with Bel7402 cells. CONCLUSION Decrease of apoptosis may be related to multidrug resistance of human hepatocellular carcinoma cells Bel7402/5-FU. The up-regulation of anti-apoptotic genes and down-regulation of pro-apoptotic genes are involved in the development of multidrug resistance in Bel7402/5-FU cells.