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| 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 | |
| 引用本文： | 何兴祥,王家.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[J].华中科技大学学报(医学版),2003,32(3):259-262. |
| 作者姓名： | 何兴祥 王家 |
| 作者单位： | 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 |
| 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，(human telomeraseRNA，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45例手术胃癌组织RNA斑点杂交检测的阳性率均为l00％，相应TRAP分析的阳性率分别为88．89％、86．67％，低于RNA斑点杂交(P＜0．05)。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，弥补了TRAP分析敏感性不足的缺点。 |
| 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 |
| 修稿时间： | 2001年12月6日 |
Preparation of Probe of hTR and Its Application for Expression of hTR in Gastric Mucosa |
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| He Xingxiang ,Wang Jialong ,Wu Jieli et al.Preparation of Probe of hTR and Its Application for Expression of hTR in Gastric Mucosa[J].Journal of Huazhong University of Science and Technology(Health Sciences),2003,32(3):259-262. | |
| Authors: | He Xingxiang Wang Jialong Wu Jieli |
| Institution: | He Xingxiang 1,Wang Jialong 2,Wu Jieli 3 et al 1 Department of Internal Medicine,Second Affiliated Hospital of Guangzhou Medical College,Guangzhou 510260 2 Department of Internal Medicine,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030 3 Department of Biochemistry,Branch Campus of First Military Medical University,Guangzhou 510315 |
| Abstract: | 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. |
| Keywords: | telomerase RNA dot blot gastric cancer telomere repeat amplification protocol |
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