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Archive > Volume 20 Issue 5 > 2013,20(5):565-568. DOI:10.3872/j.issn.1007-385X.2013.05.010 Prev Next
Basic Research
Midkine induces anoikis resistance in human hepatocellular carcinoma Hep3B cells
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Abstract:
Objective:To explore the effect of midkine (MK), a heparin-binding cytokine, on the anoikis resistance in hepatocellular carcinoma Hep3B cells. Methods: Human hepatoma-derived cell line Hep3B was kept in suspension to induce an anoikis model and treated with different concentrations of MK (0, 10, 50 and 100 ng/ml) or PBS (control group). The apoptosis of Hep3B cells was detected by flow cytometry and the expressions of apoptosis-related proteins Bcl-2 and caspase-3 were determined by Western blotting. Results:As incubation time was prolonged, the apoptotic rate of hepatocellular carcinoma Hep3B cells cultured in suspension was gradually increased. After incubation for 72 h, the apoptotic rate of Hep3B cells cultured in suspension was significantly higher than that in the adherent-cultured cells (\[38.76±423\]% vs \[6.76±1.43\]%，P<0.01). After exposure of different concentrations of MK for 24 h, the apoptotic rate of the suspension-cultured Hep3B cells was significantly lower than that of the control group and negatively correlated with the concentration of MK (r=0.951, P=0.049). Simultaneously, the expression of intracellular anti-apoptotic protein Bcl-2 was significantly increased, whereas the expression of pro-apoptotic protein caspase-3 was significantly decreased. Conclusion: MK confers enhanced anoikis resistance in hepatocellular carcinoma Hep3B cells cultured in suspension, which may be associated with the up-regulation of Bcl-2 protein and down-regulation of caspase-3 protein.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No.81272669, No.81272668), and the National Key Project for Infectious Disease (No.2012ZX10002012-10)
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