The Expression And Meaning Of ABCC4Gene In NK/T Cell Lymphoma

Objective：1．To detect the expression of ABCC4gene in NK/T cell lymphoma cell lines and tissue2．To discuss the relationship between ABCC4gene expression and the clinical therapeutic effect3．Build slow virus vector of ABCC4gene high expression and low expression, and further discuss the effect of the ABCC4gene expression quantity on NK/T cell lymphoma chemotherapyMethods：1．The expression of ABCC4gene in NK/T cell lymphoma cell lines was preliminarily detected by the technology of digital gene expression (the DGE) and the drug resistance related gene PCR gene chip.2．The expression of ABCC4/MRP4gene in NK/T cell lymphoma SNK-6, YTS cell lines and normal NK cells was detected by real-time fluorescent quantitative PCR technique.3．The protein expression levels of ABCC4/MRP4gene in110cases of tissue samples including NK/T cell lymphoma group (60cases) and rhinitis and nasal polyp group (50cases) was detected by immune histochemical method (IHC).4．According to ABCC4/MRP4gene mRNA sequence, two groups of ABCC4/MRP4gene targeting shRNA sequences were designed and the whole sequence of effective expression of ABCC4/MRP4gene was synthesized.The pCDH ABCC4, pLL3.7ABCC4-siR slow virus expression vector were also synthesized, the recombinant lentivirus Lenti ABCC4and Lenti-ABCC4–siR were prepared, and then enzyme digestion and sequencing identification were performed.5．The ABCC4/MRP4gene expression in NK/T cell lymphoma cell lines after slow virus infection was evaluated through Real time PCR and Western blot technique, at the same time, the most efficient shRNA silence was selected.6．The sensitivity of different drugs act on NK/T cell lymphoma cell lines which normally express as well as high and low express ABCC4/MRP4genes was test by CCK8experiment and flow cytometry.Results：1．Real-time fluorescent quantitative PCR (Real time PCR) technology and immune histochemical method suggested that ABCC4/MRP4genes were highly expressed in NK/T cell lymphoma cell lines and tissues, and the expression quantity was negatively correlated with the clinical curative effect.2．The slow virus expression vector pCDH ABCC4, pLL3.7ABCC4–siR were successfully builded, the recombinant lentivirus Lenti ABCC4and Lenti-ABCC4-siR1were prepared, and the ABCC4gene expression in NK/T cell lymphoma cell lines after infection of the two recombinant lentivirus respectively was effectively risen and lowed.3．Reducing ABCC4/MRP4gene expression can improve the sensitivity of NK/T cell lymphoma cell lines with different chemotherapy drugs.Conclusions：1．Multi-resistant related protein gene4(ABCC4gene) is high expressed in NK/T cell lymphoma. 2．Successful preparation of recombinant lentivirus Lenti-ABCC4and Lenti-ABCC4-siR1can effectively raise and reduce the expression of ABCC4gene in NK/T cell lymphoma cell lines.3．The lower the amount of ABCC4gene expression in NK/T cell lymphoma cell line, the more obvious the effects of chemotherapy drugs...