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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 ,弥补了TRAP分析敏感性不足的缺点
Objective To establish a method for the detection of human telomerase RNA expression. Methods cDNA probes of human telomerase RNA (hTR) were prepared and the expression of telomerase RNA in different gastric mucosa and telomerase activity were detected by RNA dot blot and telomere repeat amplification (TRAP) active. Results Human telomerase RNA cDNA probes were successfully prepared. The positive rates of RNA dot blot hybridization in 18 biopsied gastric cancer tissues and 45 surgical gastric cancer tissues were 100%. The positive rates of the corresponding TRAP assays were 88 89% and 86 6 7%, respectively, which were lower than those of RNA dot blot hybridization (P < 0 0 5). At the same time, the result of RNA dot blot showed that the expression of telomerase RNA increased with the increase of intestinal metaplasia in non-cancerous gastric tissues. Conclusion The detection of human telomerase RNA by RNA dot blot is highly sensitive and specific, which makes up for the shortcomings of the lack of sensitivity of TRAP analysis