Experimental Study On Reversion Of Malignant Phenotypes Of Bel-7402 Human Hepatocarcinoma Cell Induced By Melittin

Objective : To investigate the effect of melittin to change the malignant phenotypes of the hepatoma carcinoma cell line Bel-7402Methods: In the preliminary experiment, different doses of concentration of melittin such as 1, 2, 4, 8, 16ug/ml were used to treat Bel-7402. On the basis of the result of preliminary experiment, the final concentration of 2 g/ml and 4ug/ml were choosed as experiment groups, while All-trans retinoic acid(ATRA) was used as positive control and equal volume of of RPMI-1640 culture medium is used as negative control. After Bel-7402 was treated with melittin, detection methods were applied as followed, 1) measure the change of proliferation by MTT ; 2) light microscope and transmission electron microscope(TEM) were used to observe the morphous and biological behaviour and ConA agglutination test and clonality test were applied to measure the adhesion capability and growth potency of Bel-7402; 3) ELISA , colorimetry , bromocresol green mehod were used to determine AFP level, the activity of y - GT and ALP, ALb level respectively; 4) the change of c-myc gene expression was detect by immunohistochemical method.Result : The study showed, l)the levels of melittin concentration of 2g/ml and 4 g/mlcould both depress the proliferation of the human hepatoma carcinoma cell line Bel-7402 and mediate non-cytotoxic effect and time-effect relationship and dose-effect relationship was obvioued in a limited range; 2)the proliferation of treated Bel-7402 was put off, clonality drastically decreased and the capacity of soft agar colony formation significantly decreased; ultrastructure of treated Bel-7402 was trended to maturity under TEM; 3)AFP level and Y - GT activity both degressed in different degree; ALP activity and Albumin level of treatment groups are both significantly higher than that of contrast groups; 4) c-myc gene expression notably depressed.Conclusion: 1) Low dose of melittin could effectively induce the transformation of the malignant phenotypes of the human hepatoma carcinoma cell line Bel-7402. 2)Inductioneffect of melittin on the concentration of 2-4 g/ml is the most obvious.