谷氨酰胺酶反义基因对胃癌细胞恶性增殖的逆转作用

目的利用反义核酸技术封闭胃癌细胞的谷氨酰胺酶（glutaminase，GA）基因，观察反义GAcDNA对胃癌细胞恶性增殖能力的影响。方法构建含有GAcDNA片段的反义真核表达载体pcDNA3．0／GA，经脂质体介导转入胃癌细胞SGC-7901（命名为SGC-7901^GA），并对胃癌细胞生长曲线、细胞核增殖抗原（proliferating cell nuclear antigen，PCNA）表达、软琼脂克隆形成试验进行观察。结果与对照组比较，8GC-7901^GA在各时相点的细胞生长数目显著性降低（P〈0．05），PCNA表达较对照组显著下降，在软琼脂上克隆形成数减少。提示SGC-7901^GA生长速度减慢，增殖活性明显减弱。结论通过特异性抑制GA基因的表达，反义GAcDNA对胃癌细胞恶性增殖能力有显著抑制作用。

Inhibitory Effects of Antisense GAcDNA Oligonucleotides on Proliferation of Gastric Carcinoma Cells

Objective To investigate the effects of antisense GAcDNA on the proliferation of gastric carcinoma cells by blocking the glutaminase gene with antisense nueleotide techniques. Methods The antisense eukaryotic expression vector containing the segment of GAcDNA, named pcDNA 3. 0/GA, was constructed by standard gene recombinant techniques. Then it was transfected into gastric carcinoma cells （SGC-7901） by liposomes. These transfected cells were named SGC-7901^GA after filtration and identification. The cell growth curves and proliferating cell nuclear antigen （PCNA） and colony formation in soft agar were observed. Results The growth speed of SGC-7901^GA was significantly slower than control group （P〈0. 05）. The expression of PCNA and the ability of gastric carcinoma cells to form clones in soft agar is significantly lower in SGC-7901^GA than in control group （P〈0. 01）. Conclusion Antisense GAcDNA otigonucleotides can inhibit the proliferation of gastric carcinoma ceils.