Tim4基因定量检测方法的建立及在消化道肿瘤中的初步应用

Development of real-time RT-PCR for detection of Tim4 and its application in gastrointestinal carcinomas

目的:建立T细胞免疫球蛋白及黏蛋白域分子4(T cells immunoglobu lin-and muc in-dom ain-contain ing mole-cu le 4,Tim4)基因表达的实时荧光定量PCR(实时RT-PCR)的检测方法,探讨Tim4基因在消化道肿瘤中的表达差异。方法:构建包含Tim4基因片段的标准质粒,通过RT-PCR,实时RT-PCR等检测并比较Tim4基因在37例胃癌组织、19例结直肠癌组织与对应癌旁组织中的表达水平。结果:建立的Tim4基因定量检测方法准确可靠;在37例胃癌组织中,26例癌旁组织Tim4基因表达量高于对应癌组织(26/37),且在26例癌旁组织高表达的样本中,18例(69.2%)胃癌癌旁组织与癌组织表达比率大于2;在19例结直肠癌中12例癌旁组织表达高于对应的癌组织(12/19),其中癌旁组织高表达率(大于2)为75.0%(9/12)。结论:成功建立了检测Tim4基因的实时RT-PCR方法,Tim4基因在结直肠癌和胃癌的癌组织中表达呈低趋势,这为深入探讨Tim4基因与肿瘤的关系提供了实验基础。

Objective： To establish a method of real-time RT-PCR for T cells immunoglobulin-and mucin-domain-containing molecule 4（Tim4） and investigate its expression in colorectal cancer and gastric cancer.Methods： The standard plasmid of Tim4 gene was firstly constructed.The levels of expression of Tim4 in the tumor tissues of 37 patients with gastric cancer and 19 patients with colorectal cancer were detected by RT-PCR and real-time PCR. Results： The standard quantitative curve of Tim4 gene was generated（correlation coefficient r=0.998）.The real-time RT-PCR for Tim4 was accurate and reliable.The expression of Tim4 gene in the gastric cancer tissues（26/37） was lower than that in the matching adjacent non-cancerous tissues.The rate of higher expression of Tim4 gene（2） in adjacent non-cancerous tissues/gastric cancer tissues was 69.2%（18/26）.Likewise,12 samples expression of Tim4 gene in the colorectal cancer tissues of 19 patients were lower than that in the matching adjacent non-cancerous tissues,the rate of higher expression of Tim4 gene（2） in adjacent non-cancerous tissues was 75.0%（9/12）. Conclusion： A stable real-time RT-PCR has been successfully established for detection of Tim4 gene expression.Expression of Tim4 gene is lower in gastric cancer and colorectal cancer tissues,and provide a platform for Tim4 and cancer studies.