人端粒酶RNA的cDNA探针制备及其对胃粘膜细胞端粒酶RNA表达的检测  

Preparation of Probe of hTR and Its Application for Expression of hTR in Gastric Mucosa

作　　者：何兴祥[1] 王家 吴捷莉[3] 袁顺玉[2] 艾莉[2] 

机构地区：[1]广州医学院第二附属医院消化内科,广州510260 [2]华中科技大学同济医学院附属同济医院消化内科,武汉430030 [3]第一军医大学分校生物化学教研室,广州510315

出　　处：《华中科技大学学报（医学版）》2003年第3期259-262,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

摘　　要：目的　建立人端粒酶RNA表达的检测方法。方法　制备人端粒酶RNA ,(humantelomeraseRNA ,hTR)的cDNA探针 ,分别应用RNA斑点杂交与端粒重复序列扩增法 (TRAP)分析检测不同胃粘膜的端粒酶RNA的表达与端粒酶活性。结果　人端粒酶RNA的cDNA探针制备成功。 18例活检胃癌组织及 4 5例手术胃癌组织RNA斑点杂交检测的阳性率均为 10 0 % ,相应TRAP分析的阳性率分别为 88 89%、 86 6 7% ,低于RNA斑点杂交 (P <0 0 5 )。同时RNA斑点杂交结果提示在非癌胃组织中随着肠化程度增高人端粒酶RNA表达也增强。结论　RNA斑点杂交检测人端粒酶RNA ,具有高度的敏感性和特异性 。Objective To establish the method for detection of the expression of human telomerase RNA(hTR). Methods The probe of hTR was obtained by RT-PCR using total RNA of HeLa cells. The expression of hTR and telomerase activity in gastric mucosa was measured by RNA dot blot and telomere repeat amplification protocol (TRAP) assay , respectively. Results RNA dot blot was accomplished using the probe of hTR. The result showed that the positive rate was 0, 0, 100 % and 100 % in 10 cases of normal gastric mucosa, 46 cases of chronic superficial gastritis mucosa, 18 cases of biopsy specimens and 45 cases of resected gastric carcinoma specimens, respectively. In contrast, the positive rate of TRAP was 0, 0, 88.89 % and 86.67 % in those samples, respectively (P<0.05). At the same time, RNA dot blot revealed that the expression of hTR was increased with intensified intestinal metaplasia in gastric mucosa. Conclusion RNA dot blot has high sensitivity and specificity to detect the expression of hTR, and remedies defect of low sensitivity of TRAP.

关 键 词：胃癌 端粒酶 核糖核酸 端粒重复序列扩增法 RNA斑点杂交法 

分 类 号：R735.2[医药卫生—肿瘤]