An Experimental Study On The Effect Of Using Antisense Nucleotide Technique To Inhibit The Expression Of Gulutaminase Gene

Objectives: To estimate the inhibition effect on tumor growth after the glutaminase gene was blocked with antisense nucleotide technique, we injected the gastric carcinoma cells transfected with plasmid containing the C' distal end antisense sequence of glutaminase gene into nude mouse body and compared its proliferation and neoplasia capability with control gastric carcinoma cell.To provid a new academic basis for the gene therapy of tumor.Methods: The gastric carcinoma cells had been successfully transfected with plasmid containing the C' distal end antisense sequence of glutaminase gene and called SGC-7901GA in previous researches. The SGC-7901GA cells and control cells SGC-7901Neo were cultured in vitro,and then were respectively injected into the endermic tissue of 6-weeks-old BALB/C node mice to create the animal models of tumor. The neoplasia and growth of tumor were observed. Use MTT method to detect the difference of growth capability of two kinds of cells in vitro. The expression of proliferating cell nuclear antigen(PCNA) in tumor cells was detected by immunohistochemistry (S-P method). The apoptosis of tumor was detected by terminal deoxynucleotidyl transferase-mediated nick end labeling technique(TUNEL). The content of glutaminase mRNAs in tumor tissue was measured by reverse transcription polymerase chain reaction (RT-PCR) technique.Results:1. The growth curve obtained with MTT method showed that the SGC-7901GA cells' proliferation speed is significantly slower than that of control group.2. The neoplasia time of experimental group is significantly later than that of control group after injected cells at same concentration. The tumor's growth speed of experimental group is also significantly slower than that of control group. The tumor's mean weight of experimental group is significantly less than that of control group after ablated.3. The tumor's proliferate index of experimental group is significantly less than thatof control group. While it's apoptosis index is significantly higher than control group.4. Glutaminase mRNAs in tumor tissue of experimental group is significantly less than that in control group.Conclusions: Blocking the gene of glutaminase in gastric carcinoma with antisense nucleotide technique can inhibit the expression of glutaminase protein and decrease the activity of glutaminase. It can hinder gastric carcinoma's utilization of glutamine, so as to inhibit the proliferation of tumor and promote the apoptosis of tumor cells. It may be a effective therapy of tumor by blocking the gene of glutaminase with antisense nucleotide technique.