Study Of Apoptosis Induced By Adriamycin And Apoptosis-associated Genes And "Killer Factors" On Hepatoma Cells

Apoptosis was closely related to tumors. The study on apoptosis will be expoloiting a new way to overcome the tumors, potentially. However, the molecular mechanisms of apoptosis is unknown up to now. Therefore, it is important and significant to further investigate and explore the molecular mechanisms of apoptosis on tumor cells. Hepatoma is one of the common malignant tumors in our country. Aimed at further exploring and demonstrating the molecular mechanisms of apoptosis on hepatoma, the apoptosis was studied in more detail on hepatoma cells by a serise of techniques in molecular biology and molecular pathology.This study consisted of four major parts, which were introduced as follows:Firstly, the apoptosis of human cultured hepatoma cells was induced by adriamycin and the model of apoptosis was established in hepatoma cells. The results showed that the induction of apoptosis was significantly inhibited by actinomycin D at 0.5 ug/ml in the hepatoma cells treated with adriamycin at 20 umol/L,which suggested that some new large molecular proteins were synthesized actively during the process of apoptosis in hepatoma cells.The morphological changes of hepatoma cells were accorded with the characteristics of apoptotic cells at the lever of acridine orange staining, light microscopy and electron microscopy. The apoptosis-associated DNA fragmentation occured, referred to as a 'DNA ladder'. The typical apoptotic peak was observed by flow cytometry in hepatoma cells.The apoptosis of hepatoma cells occurred mainly in G1 phase of cell cycles. Therefore, the establishment of model of apoptosis in hepatoma laid a foundation for the further study of apoptosis on hepatoma.Secondly, in order to investigate the influence of wild-type p53 gene transferred to hepatoma cells, the wild-type p53 gene was transfected into HCC-9204 hepatoma cell line. The endogenous p53 gene of HCC-9204 cell line was detected by polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) analysis and immunohistochemistry methods. The results showed that the mutation in exon 7 of p53 gene was found in HCC-9204 hepatoma cell line. And also p53 proteins were positive immunohistochemically in hepatoma cells,which further proved that the endogenous p53 gene of HCC-9204 hepatoma cells existed in the pattern of...