Hu Li-Hua, Chen Feng-Hua, Li Yi-Rong, Wang Lin
Department of Transfusion, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China.
World J Gastroenterol. 2004 Dec 1;10(23):3514-7. doi: 10.3748/wjg.v10.i23.3514.
To set up a real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-PCR) assay, to detect human telomerase reverse transcriptase (hTERT) messenger RNA in gastric carcinomas, and to evaluate quantitative determination of hTERT mRNA in the diagnostic value of gastric carcinomas, and to analyze the correlation between the expression level of hTERT mRNA and clinicopathological parameters in patients with gastric cancer.
A real-time quantitative RT-PCR (RQ-PCR) based on TaqMan fluorescence methodology and the LightCycler system was used to quantify the full range of hTERT mRNA copy numbers in 35 samples of gastric carcinomas and corresponding adjacent non-cancerous tissues. The normalized hTERT (NhTERT) was standardized by quantifying the number of GAPDH transcripts as internal control and expressed as 100X (hTERT/GAPDH) ratio. Variables were analyzed by the Student's t-test, chi2 test and Fisher's exact test.
NhTERT from gastric carcinomas and corresponding adjacent non-cancerous tissues was 6.27+/-0.89 and 0.93+/-0.18, respectively (t = 12.76, P<0.001). There was no significant association between gastric cancer hTERT mRNA expression level and patient's age, gender, tumor size, location and stage (pTNM), but a significant correlation was found between hTERT mRNA expression level in gastric carcinomas and the degree of differentiation.
Quantitative determination of hTERT mRNA by RQ-PCR is a rapid and sensitive method. hTERT might be a potential biomarker for the early detection of gastric cancer.
建立实时荧光定量逆转录-聚合酶链反应（RT-PCR）检测方法，检测胃癌组织中人端粒酶逆转录酶（hTERT）信使核糖核酸（mRNA），评估hTERT mRNA定量检测在胃癌诊断中的价值，并分析胃癌患者hTERT mRNA表达水平与临床病理参数的相关性。
采用基于TaqMan荧光法和LightCycler系统的实时定量RT-PCR（RQ-PCR），对35例胃癌组织及相应癌旁非癌组织中hTERT mRNA的拷贝数进行全范围定量分析。以甘油醛-3-磷酸脱氢酶（GAPDH）转录本数量作为内参对hTERT进行标准化处理，以100×（hTERT/GAPDH）比值表示。采用Student's t检验、χ²检验和Fisher精确检验对变量进行分析。
胃癌组织及相应癌旁非癌组织的标准化hTERT（NhTERT）分别为6.27±0.89和0.93±0.18（t = 12.76，P<0.001）。胃癌hTERT mRNA表达水平与患者年龄、性别、肿瘤大小、部位及分期（pTNM）无明显相关性，但与胃癌组织的分化程度有显著相关性。
RQ-PCR定量检测hTERT mRNA是一种快速、灵敏的方法。hTERT可能是胃癌早期检测的潜在生物标志物。
