摘要
目的:胃癌下调新基因CAll的细胞定位及其生物学功能的研究。 方法:地高辛标记CAll原位正常胃黏膜组织mRNA杂交。将CAll构建至pcDNA3.1/Myc-His(-)A载体,将其及空载pcDNA3.1/Myc-His(-)A载体稳定转染胃癌7901细胞系,Western blot证实CAll转染细胞CAll融合蛋白表达,行生长曲线及MTT检测,形态学分析结果。 结果:NBT/BCIP显色表明,CAll位于正常胃黏膜上皮细胞胞内,CAll稳定转染胃癌7901细胞系表达CAll融合蛋白,生长曲线及MTT检测均表明,CAll显著抑制胃癌7901细胞的生长(72h,0.379±0.019A us 0.467±0.021 A,P<0.01)。较对照组空载转染7901细胞,CAll稳定转染胃癌7901细胞系形态不规则,核糖体、胞膜纤毛明显减少。 结论:CAll位于正常胃黏膜细胞,能显著抑制胃癌细胞的生长,可能为一功能强大的候选抑癌基因。
AIM:To demonstrate the location of CAll in gastric mucosal cells, and to confirm the function of CAll in gastric cell lines 7901.METHODS: CA11 mRNA was detected in gastric mucosal cells by mRNA hybridization. CAll was cloned into vector pcDNA3. 1/Myc-His (-) A and steadily transfected into gastric cell lines 7901 by lipofectamin. Control vector pcDNA3.1/Myc- His (-) A was also transfected into the cell lines. CAll fusion protein expression was detected by Western blot. Growth curve, MTT test and a morphological analysis were performed in this study.RESULTS: CA11 was located in gastric mucous epithelial cells. Gastric cell lines 7901 steadily transfected by CA11 expressed CA11 fusion protein, and showed a marked decrease in growth rate by growth curve and MTT test (72 h, 0.379 ±0.019 A vs 0.467 ±0.021 A, P<0.01). Irregular shapes, less ribosome or cilia on cell membrane were observed in gastric cell lines 7901 transfected by CA11.CONCLUSION: CA11 is located in gastric mucous epithelial cells. CA11 can markedly suppress growth of gastric cell lines 7901, and may be a candidate of suppressor genes with potential functions.
出处
《世界华人消化杂志》
CAS
2002年第5期525-529,共5页
World Chinese Journal of Digestology
