IL-10对稳定转染HBx基因的HL-7702细胞增殖的影响

发布时间：2018-04-26 07:31

本文选题：HBx + HL-7702　；参考：《福建医科大学》2014年硕士论文

【摘要】：目的：探讨HBx基因对HL-7702细胞增殖的作用，研究分析IL-10对稳定转染HBx基因的HL-7702细胞增殖的作用以及其与CDKN1B mRNA表达的关系。方法：体外培养HL-7702细胞、HL-7702/MOCK细胞与HL-7702/HBx细胞；CCK8法测定HBx基因对HL-7702细胞增殖的作用及IL-10对稳定转染HBx基因的HL-7702细胞增殖的作用；流式细胞术测定IL-10对HL-7702/HBx细胞凋亡的影响；RT-PCR法测定HBx基因的对HL-7702细胞表达CDKN1B mRNA的作用及IL-10对HL-7702/HBx表达CDKN1B mRNA的作用。结果： CCK8法显示HL-7702/HBx细胞比HL-7702细胞和HL-7702/MOCK细胞增殖速度明显加快，，HL-7702细胞和HL-7702/MOCK细胞增殖无差别；80ng/ml IL-10作用24小时可抑制HL-7702/HBx细胞增殖，而对HL-7702细胞、HL-7702/MOCK细胞增殖无影响；80ng/ml IL-10作用24小时对HL-7702细胞、HL-7702/HBx细胞、HL-7702/MOCK细胞凋亡均无影响；HL-7702/HBx细胞CDKN1B mRNA表达量较HL-7702细胞和HL-7702/MOCK细胞降低，而HL-7702细胞和HL-7702/MOCK细胞CDKN1BmRNA表达量无差别；80ng/ml IL-10作用24小时后HL-7702/HBx细胞较HL-7702/HBx空白组CDKN1B mRNA表达量上调。结论：HBx基因可促进HL-7702细胞增殖，IL-10可抑制HL-7702/HBx细胞增殖而对其凋亡无影响，HBx基因可能通过下调HL-7702/HBx细胞CDKN1B mRNA表达量而促进细胞增殖，IL-10可能通过上调HL-7702/HBx细胞CDKN1B mRNA表达量而抑制细胞增殖。
[Abstract]:Aim: to investigate the effect of HBx gene on the proliferation of HL-7702 cells, and to investigate the effect of IL-10 on the proliferation of HL-7702 cells stably transfected with HBx gene and its relationship with the expression of CDKN1B mRNA. Methods: the effect of HBx gene on the proliferation of HL-7702 cells and the effect of IL-10 on the proliferation of HL-7702 cells stably transfected with HBx gene were determined by HL-7702 cell line HL-7702 / mock and HL-7702/HBx cell line CCK8 in vitro. The effect of IL-10 on apoptosis of HL-7702/HBx cells was determined by flow cytometry. The effect of HBx gene on CDKN1B mRNA expression in HL-7702 cells and the effect of IL-10 on HL-7702/HBx CDKN1B mRNA expression were determined by reverse transcription-polymerase chain reaction (RT-PCR). Results: CCK8 assay showed that the proliferation rate of HL-7702/HBx cells was significantly faster than that of HL-7702 cells and HL-7702/MOCK cells. The proliferation of HL-7702 cells and HL-7702/MOCK cells was similar to that of HL-7702/MOCK cells. The proliferation of HL-7702/HBx cells was inhibited by 80 ng / ml IL-10 for 24 hours. However, there was no effect on the proliferation of HL-7702 / mock cells in HL-7702 cells. The expression of CDKN1B mRNA in HL-7702 / mock cells was significantly lower than that in HL-7702 cells and HL-7702/MOCK cells after 24 hours of exposure to 80 ng / ml IL-10. The apoptosis of HL-7702 / MoCK cells in HL-7702 / HL-7702 / HBX cells was not affected by HL-7702 / Mock cells. However, there was no difference in the expression of CDKN1BmRNA between HL-7702 cells and HL-7702/MOCK cells. After 24 hours of treatment with 80 ng / ml IL-10, the expression of CDKN1B mRNA in HL-7702/HBx cells was higher than that in HL-7702/HBx control group. Conclusion the cell proliferation of HL-7702 cells can be promoted by 1: HBX gene. IL-10 can inhibit the proliferation of HL-7702/HBx cells, but it has no effect on the apoptosis of HL-7702/HBx cells. It may promote the proliferation of HL-7702/HBx cells by down-regulating the expression of CDKN1B mRNA. IL-10 may inhibit the proliferation of HL-7702/HBx cells by upregulating the expression of CDKN1B mRNA.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R512.62

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