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Objective To observe the effects of AMPK activation on cancer cell apoptosis and to investigate the underlying mechanism and its significance to cancer therapy.Methods Human cervical cancer HeLa-S3 cells were divided into six groups:DMSO control,5 μmol/L CD437,CD437+Compound C,CD437+AICAR,Compound C,AICAR.Apoptosis rates in every groups induced by CD437 and other factors were determined by FACS and confirmed with clave Caspase-3 expression by Western blot analysis.Finally,apoptosis rates were detected in HeLa-S3 cells induced by the above factors after wt-LKB1 reintroduction by plasmid transfection.Results FACS results showed that treatment with 5 μmol/L CD437 for 8 hours could induce Hela-S3 cells apoptosis by (87.42±5.95) ％.Co-incubation with 20 μmol/L compound C enhanced apoptotic effect of CD437 by (86.42±5.95) ％,while co-incubation with 2 mmol/L AICAR markedly reduced cell apoptosis induced by CD437 [(51.04±8.26) ％] (P ＜ 0.05).Re-introduction of wt-LKB1 by plasmid transfection attenuated the protective effect of AICAR in CD437 induced apoptosis.Conclusion Endogeneous AMPK activation exerts a protective effect in HeLa-S3 ceils.This effect could be attenuated by re-introduction of wt-LKB1.
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Objective The prokaryotic expression vector of NADP(H)-dependent retinol dehydrogenase/reductase(NRDR)B1 was constructed for the detection of NRDRB1 protein,and to prepare its polyclonal antibodies,in order to lay the foundation to study the function of NRDRB1.Methods The coding region of NRDRB1 was constructed to the Gateway-based expression vector(pDEST 14),which was transformed into the Escherichia coli(BL21-AI)for the native protein expression.Overexpression of the recombinant was induced at mid-log growth phase of BL21-AI(A600=0.6)using 0.2% L-arabinose.After supersonication the inclusion bodies of NRDRB1 were purified.New Zealand rabbits were immunized with NRDRB1 as the immunogen,which was recovered from SDS-PAGE gel and subscapsularly injected.The titer of the antiserum was determined by dot blot assay.The antibody was purified by HiTrap Protein G column,and its activity and specificity were assessed by Western blotting and immunohistochemistry.Results The prokaryotic expression vector pDEST 14 with NRDRB1 was constructed.The constructs were sequenced by dideoxynucleotide method.NRDRB1 was overexpressed in strain BL2l-AI.The concentration of recovered NRDRB1 was 0.42mg/ml with a recovery rate of 52.3%.All the immunized rabbits produced high-titer antisera after the second booster.The titer of the antiserum was 1∶2 000 with a detection limit of 6.4ng NRDRB1.The purified antibody had a high specificity.Conclusion The present study provides an effective method of preparing polyclonal antibody against NRDRB1.The purified NRDRB1 native protein and the specific polyclonal antibody of NRDRB1 would be valuable for the study on the biological function of NRDRB1.
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Retinoid acid and its derivatives have shown promising perspective in clinical use and lab research on the leukemia and other solid tumor cells.Some of these compounds have a stronger apoptotic potential,a lower level of cytotoxicity and a better pharmacokinetic profile than all-trans retinoic acid.The apoptosis pathways induced by these compounds are different from traditional p53-dependent pathway which recruited by many chemotherapeutics.Due to its specific molecular mechanism,these compounds could induce apoptosis in retinoic acid-resistant or multi-drug-resistant tumor cells.This paper reviews the current research on apoptosis induced by analogues of retinoid acid.
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Objective: The antitumor mechanism of Pleurotes sapidus on 4 kinds of tumor cell lines was studied. The active components of its ethanol extracts were studied. Methods:Using techniques of cell culture in vitro, phytochemical methods, FACS and RT-PCR, expressions of p53 and fas gene of four tumor cell lines treated with Pleurotas sapidu were analysed. Results:The apoptosis rates were remarkably increased after treated by ethanol extracts (0.1g/ml) and water extracts (0.1g/ml) for 12 h and 24 h respectively. The expression of P53 protein in the groups of ethanol and water extracts were significantly increased, and the gene expressions of p53 and fas were aslo highly up-regulated. Conclusion:The antitumor effect of Pleurotas sapidus on tumor cell lines in vitro is evident, and its mechanisms are probably associated with the regulation of apoptosis by inducing the gene expressions related to apoptosis.
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Objective: To study the anticarcinogenic effects of raisin grape produced in Turpan in vitro; to determine the content of the components related to anticarcinogenesis.Methods: The effect of Turpan raisin grape on the growth of four tumor cell lines and one normal cell line was observed. The survival rate and protein content of cells were detemined. Four components in the Turpan raisin grape were measured, including vitamin C, polysaccharide, bioflavonoids and selenium.Results: The extracts of Turpan raisin grape significantly inhibited the growth of four tumor cell lines (P