| Objective:To observe the inhibition effects of adriamycin andNS-398 on protein of survivin expression and mechanisms of apoptosis inhepatocellular carcinoma cell line HepG2.Methods: HepG2 were treated with adriamycin and NS-398 in variousconcentrations and then MTT cell proliferation assay was used to observedthe inhibition of HepG2 cells and the expression of survivin protein andmRNA were detected by immunofluorescence and RT—PCRrespectively.Results: (1) Adriamycin can significantly inhibit the cells increase,and the inhibition was enhanced by combining with NS-398; (2) Thesurvivin proteinum and mRNA was highly expressed in HepG2 cells andthe predominant localization of survivin proteinum was cytoplasm beforeHepG2 were interfered with drugs。The quantity of survivin fluorescencewas 68.971±1.155 and the survivin mRNA/GAPDH mRNA ratio was0.480±0.013; (3) Expression of surviving protein was reduced moremarkedly in cells that treated with adriamycin, the quantity of survivinfluorescence was 47.629±0.864 and the survivin mRNA/GAPDH mRNA ratio was 0.748±0.0138; when treated with a combination of adriamycin andNS-398 the quantity of survivin fluorescence was 27.5434-0.747, and thesurvivin mRNA/GAPDH mRNA ratio was 0.480±0.013.(P＜0.05)Conclusion: (1) The survivin proteinum and mRNA was highlyexpressed in HepG2 cells and the predominant localization of survivinproteinum was cytoplasm, there were less survivin proteinum in nucelus;(2) adriamycin can significantly inhibit the cell increase and it appears dosedependent and the inhibition was enhanced by combining with NS-398; (3)adriamycin can downregulate the expression of survivin proteinum andmRNA in HepG 2 cells and this effect can be enhanced by combination ofthese two drugs; (4) adriamycin and NS-398 may induce HepG2 cellsapoptosis through the downregulation of survivin protein expression andthus exert their anti—cancer effect. Their combination may enhance theiranti—tumor effect and reduce the dosage of adriamycin.
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