Effects of Glutaminase Antisense Gene on Apoptosis of Transplanted Gastric Carcinoma Cells in Nude Mouse_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

LIU Menggang , YANG J untao , LIU Hongming , YUAN Tao , GU Hongguang

Department of Hepatobiliary Surgery , Daping Hospital , The Third Military Medical University, Chongqing 400042, China;

Corresponding author：

Keywords：

Gast ric carcinoma，　　Antisense nucleotide，　　Glutaminase，　　Cell apoptosis

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Objective 　To study the effects of glutaminase (GA) gene blocked by antisense nucleotide on apoptosis of transplanted gastric carcinoma cells in nude mice. Methods 　The plasmid containing antisense sequence of GA gene was trans-fected into gastric carcinoma cells , then the cells were injected to endermic tissue of nude mice to create animal models of gastric carcinoma. Apoptosis of tumor cells was detected by terminal deoxynucleotidyl transferase2mediated nick end labeling (TUNEL) method. The expression of GA mRNA in tumor tissue was measured by reverse transcription polymerase chain reaction (RT2PCR) technique. Results 　After the successful transfection of plasmid containing antisense sequence of GA gene into gastric carcinoma cells , the tumor’s growth speed decreased , apoptosis of tumor cells increased , and the expression of GA mRNA also decreased. Conclusion 　The antisense gene of GA could inhibit the expression of GA gene and significantly increase the apoptosis of gastric carcinoma cells.

Citation： LIU Menggang,YANG J untao,LIU Hongming,YUAN Tao,GU Hongguang. Effects of Glutaminase Antisense Gene on Apoptosis of Transplanted Gastric Carcinoma Cells in Nude Mouse. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2008, 15(1): 4-7. doi: Copy

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1. Medina MA,Sanchez-Jiménez F,Mrquez J,et al.Relevance of glutamine metabolism to tumor cell growth[J].Mol Cell Biochem,1992; 113(1)∶1.
2. Medina MA.Glutamine and cancer[J].J Nutr,2001; 131(9 Suppl)∶2539S.
3. 刘宏鸣,杨俊涛,顾红光.大鼠谷氨酰胺酶反义真核表达载体的构建及鉴定[J].医学研究生学报,2003; 16(7)∶487.
4. Simpson DA,Feeney S,Boyle C,et al.Retinal VEGF mRNA measured by SYBR green Ⅰ fluorescence:A versatile approach to quantitative PCR[J].Mol Vis,2000; 6∶178.
5. Zhang WW,Fang X,Branch CD,et al.Generation and identification of recombinant adenovirus by liposome-mediated transfection and PCR analysis[J].Biotechniques,1993; 15(5)∶868.
6. Medina MA,Sánchez-Jiménez F,Quesada AR,et al.Effect of palmitate,acetate and glucose on glutamine metabolism in Ehrlich ascites tumor cells[J].Biochimie,1988; 70(6)∶833.
7. Pérez-Gómez C,Campos-Sandoval JA,Alonso FJ,et al.Co-expression of glutaminase K and L isoenzymes in human tumour cells[J].Biochem J,2005; 386(Pt 3)∶535.
8. Zacharias DP,Lima MM,Souza AL Jr,et al.Human cutaneous melanoma expresses a significant phosphate-dependent glutaminase activity:a comparison with the surrounding skin of the same patient[J].Cell Biochem Funct,2003; 21(1)∶81.
9. Turner A,McGivan JD.Glutaminase isoform expression in cell lines derived from human colorectal adenomas and carcinomas[J].Biochem J,2003; 370(Pt 2)∶403.
10. Aledo JC,Segura JA,Medina MA,et al.Phosphate-activated glutaminase expression during tumor development[J].FEBS Lett,1994; 341(1)∶39.
11. Aledo JC,de Pedro E,Gómez-Fabre PM,et al.Changes in mRNAs for enzymes of glutamine metabolism in the tumor-bearing mouse[J].Anticancer Res,2000; 20(3A)∶1463.
12. 陈道达,郑海.胃癌的基因治疗现状、问题和展望[J].中国普外基础与临床杂志,2002; 9(3)∶141.
13. 朱金明,余佩武.胃癌生物治疗现状与展望[J].中国普外基础与临床杂志,2004; 11(2)∶127.
14. 陆芬英,张尤历.胃癌基因治疗研究进展[J].医学综述,2007; 13(3)∶191.
15. 刘孟刚,杨俊涛,刘宏鸣.GA反义基因对裸鼠胃癌移植瘤的抑制作用[J].消化外科,2005; 11(4)∶430.
16. Fulda S,Debatin KM.Death receptor signaling in cancer therapy[J].Curr Med Chem Anticancer Agents,2003; 3(4)∶253.
17. Oka T,Yoshikawa M,Kobayashi T,et al.Effects of antisense oligodeoxynucleotides against opioid receptors on the receptor mRNA contents[J].Life Sci,2001; 68(19-20)∶2221.
18. Franek F,Fismolova I,Eckschlager T.Antiapoptotic and proapoptotic action of various amino acids and analogs in starving MOLT-4 cells[J].Arch Biochem Biophys,2002; 398(1)∶141.
19. Ghosh S,Roy S,Banerjee M,et al.Modulation of tumor induced angiogenesis in Ehrlich ascites tumor[J].J Exp Clin Cancer Res,2004; 23(4)∶681.
20. Lora J,Alonso FJ,Segura JA,et al.Antisense glutaminase inhibition decreases glutathione antioxidant capacity and increases apoptosis in Ehrlich ascitic tumour cells[J].Eur J Biochem,2004; 271(21)∶4298.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Effects of Glutaminase Antisense Gene on Apoptosis of Transplanted Gastric Carcinoma Cells in Nude Mouse

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