摘要
目的分析胃癌发生各阶段组织中增殖诱导配体(APRIL)及其受体mRNA的表达水平.方法在APRIL及其受体基因的高保守区设计相应引物和荧光探针,实时检测PCR产物的荧光强度,根据标准品建立标准曲线,由软件自动计算出待测样本中靶基因mRNA的准确含量,并以靶基因和内参β2微球蛋白(β2M)mRNA含量比值作为评价靶基因表达水平的指标.结果采用实时荧光定量PCR(RFQ-PCR)检测靶基因mRNA含量的线性范围为101～109pg/ml,批内和批间重复性测定的变异系数(cv)分别为6.52%～12.02%和8.76%～14.16%.APRIL在肠上皮化生、异型增生和胃癌组织中的表达水平显著高于正常胃黏膜(P＜0.05),在胃癌组织中的表达水平又显著高于肠上皮化生和异型增生(P＜0.05),而其受体B细胞成熟抗原(BCMA)和穿膜蛋白活化物(TACI)在各种胃黏膜病变之间表达水平差异无统计学意义(P＞0.05).结论RFQ-PCR检测APRIL及其受体mRNA含量,具有较好的灵敏度和重复性.APRIL在胃癌病变演进过程中呈现累积和渐进趋势,可能在胃癌发生、发展过程中起重要作用,有可能成为胃癌早期诊断和抗癌治疗的靶分子.肿瘤组织可能还表达APRIL未知受体.
Objective To investigate the role of the expression of a proliferation inducing ligand （APRIL） and its receptors in the gastric carcinogenesis. Methods The real-time fluorescence quantita rive polymerase chain reaction （RFQ-PCR） was used to detect expression of APRIL and its receptors. Based on the standard curves, the quantitative levels of target genes in tissue samples were determined by using software, and the results were presented as the ratios of mRNA levels of target genes to β2 micro gluobulin（β2M）. Results The detection linear range of RFQ-PCR was 10^1 -10^9 pg/ml and the coefficient of variation values for both intrakexperimental and inter experimental reproducibility ranged 6.52%- 12.02% and 8.76%- 14. 16%, respectively. The expression levels of APRIL in the tissue of intestinal metaplasia , dysplasia and gastric cancer were significantly higher than those in normal tissue（P〈0.05）, and the expression level of APRIL in the tissue of gastric cancer was also significantly higher than those in the tissue of intestinal metaplasia and dysplasia （P〈0.05） . Besides, there were no significant differences on expression levels of B cell maturation antigen（BCMA）, transmembrane activator, calcium modulator and cyclophilin ligand interactor （TACI） within different groups（P〉0.05 , respectively）. Conclusions The present study indicated that RFQ-PCR had satisfied sensitivity and reproducibility in quantitative measurement of APRIl. and its receptors. APRIl. may play an important role in the development and progress of gastric cancer and could be established as a target molecule for early diagnosis and anti-cancer therapy. Besides, there maybe some unknown receptors of APRIl. expressed on tumor tissue.
出处
《中华消化杂志》
CAS
CSCD
北大核心
2005年第8期476-479,共4页
Chinese Journal of Digestion
