Studies on Effects of HC56 Gene on Lymphoma and Leukemia Raji ...

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Cancer Research

[Abstract]

Objective: The HC56 gene was cloned from the human chromosome 17pl3.3 with apositional cloning technique by the National Key Laboratory for Oncogene and Related Gene, Shanghai Cancer Institute. In our previous research, we found HC56 gene rearrangement in Lymphoma /Leukemia Cell line, and the malignant phenotype being significantly prohibited in Raji cells transfected with exogenous HC56 gene. In our present studies, we used the Raji cells stably expressing exogenous HC56 gene as experiment model, incubated with the chemotherapy drug in order to investigated whether the HC56 gene could increase the sensitivity to the chemotherapy drug and to investigate the mechanism of possible synergism.Methods :( 1) exogenous HC56 gene was transfected into Raji cells by medium with liposome and the cells were screened with G418, and expression of exogenous HC56 gene products was verified by western blot. (2)The Raji cells stably expressing exogenous HC56 gene exposured to various concentration of Daunorubicin (DNR) , Etoposide (VP16) and Vincristine (VCR) for 24 hours, the cell viability was measured by trypan blue dye exclusion test, and the 50% inhibitory concentration (IC50) were calculated. (3)The Raji Cells stably expressing exogenous HC56 gene exposured to 10 u g/ml of VP16, the cell apoptosis was observed by means of flow cytometry, Giemsa staining and Hoechst 33258 fluorescence staining.Results: (1) The expression level HC56 gene in the Raji cells transfected with exogenous HC56 gene was obviously higher than the control cells transfected with3empty vector or without any transfection.(2)Cells viability and the level of IC50 in stably expressing exogenous HC56 gene were lower than that in the control cells transfected with vector or the Raji cells without any transfection 24 hours after exposure to various concentrations of DNR and VP16.(3)Compared to the control cells transfected with vector or without any transfection, the Raji cells stably expressing exogenous HC56 gene cell viability was not dramatically inhibited, and the level of ICso was not significantly decreased after exposured to various concentrations of VCR for 24 hours. (4)Apoptosis rate of the Raji cells which stably expressing exogenous HC56 gene was only about 4% higher than the other two control groups of Raji cells 24 hours after exposure to VP16(10u g/ml), by means of flow cytometry, Giemsa staining and Hoechst 33258 fluorescence staining .Conclusion: HC56 gene could increase the Raji cells sensitivity to DNR and VP16,except VCR. (2)The mechanism of this synergism between HC56 gene and VP16 maybe not through inducing apoptosis.

Title: Studies on Effects of HC56 Gene on Lymphoma and Leukemia Raji Cell?s Sensitivity of Chemotherapy

Category: Ovarian Cancer

Filename: Studies on Effects of HC56 Gene on Lymphoma and Leukemia Raji Cell?s Sensitivity of Chemotherapy.pdf

Pages: 105

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