GFRa1不同剪接变异体mRNA在胃癌及结肠癌组织中的表达

The mRNA Expression of Different Splice Variants of GFRa1 in Gastric Cancer and Colon Cancer Development Process

为检测正常胃粘膜及胃癌组织和正常结肠及结肠癌组织中GFRa1 m RNA表达量,以及两种剪接体GFRa1a和GFRa1b的m RNA含量。初步明确胃及结肠组织中GFRa1剪接体存在形式以及与胃癌、结肠癌发生的相关性。采用收集23例正常胃粘膜/胃炎组织、20对胃癌及其切缘组织和6例非癌患者正常结肠活检组织、20对结肠癌及其切缘组织的方法。选择并提取结肠癌细胞系RKO,正常人胃粘膜上皮细胞系GES-1和人胃癌细胞SGC-7901这3种细胞系的RNA。以Alu m RNA为内参照,用定量RT-PCR的方法对胃组织、结肠组织以及所选的细胞系中GFRa1总m RNA含量进行分析。运用DHPLC技术区分GFRa1不同的剪接体,定量测定两种转录本剪接体的比值。结果显示,胃及结肠组织中,与正常及癌组织相比,切缘组织GFRa1总m RNA、剪接体a及剪接体b的m RNA相对拷贝数均显著增加(P<0.05)。所有组织中GFRa1剪接体b的m RNA相对拷贝数均高于剪接体a。细胞系RKO、GES-1和SGC-7901均检测到GFRa1剪接体b的表达,而未检测到剪接体a。由此可知,胃和结肠组织以及相应的细胞系中GFRa1主要以剪接体b的形式存在;GFRa1剪接体b的高表达可能在胃及结肠组织癌变过程发挥作用。

To detect the total quantity of GFRal mRNA expression in normal gastric mucosa and Gastric cancer tissue and normal colon and colon cancer tissue,as well as the two splice variants of GFRal named GFRala and GFRalb.Preliminary clear what kind of forms spliceosome exists in gastric cancer and colon cancer tissue,and discuss the influence of the existence form on the development process of colon cancer and gastric cancer. 23 cases normal gastric mucosa/gastritis tissues,20 cases gastric cancer and its cut edge group and 6 cases normal colonic tissues,20 cases colon cancer tissues and its cutting edge were collected respectively.The RNA of colon cancer cell lines RKO,RNA normal human gastric epithelial cell lines GES-1 and human gastric cancer cell lines SGC-7901 were selected and extracted.Take Alu mRNA for internal reference,GFRal mRNA content were analyzed in stomach tissue and colon tissue by the method of quantitative RT-PCR; The product of RT-PCR was analyzed by DHPLC,and further to distinguish the different spliceosome of GFRal,quantitative determinate the ratio of the two kinds of the spliceosome transcription. In stomach tissues and colon tissues ,the mRNA relative copy numbers of GFRala and GFRalb and total mRNA relative copy numbers of GFRal in cut edge tissue were significantly higher than that of normal gastric tissues/gastritis and cancer tissues （P〈0.05）.Besides,mRNA relative copy numbers of GFRalb were higher than spliceosome GFRala.The expression of GFRal spliceosome b of cell lines RKO,GES-1 and SGC-7901 were all detected.In the stomach and colon tissue,GFRal mainly exists in the form of spliceosome b; the high expression of GFRalb probability paly a role in stomach and colon carcinogenesis process.