通过对石蜡切片中解剖组织进行实时逆转录聚合酶链反应，定量分析肾上皮性肿瘤中G250 mRNA的表达。

Quantification of G250 mRNA expression in renal epithelial neoplasms by real-time reverse transcription-PCR of dissected tissue from paraffin sections.

作者信息

Bismar Tarek A, Bianco Fernando J, Zhang Hongquan, Li Xingli, Sarkar Fazlul H, Sakr Wael A, Grignon David J, Che Mingxin

机构信息

Department of Pathology, Harper University Hospital, Wayne State University, 3990 John R., Detroit, MI 48201, USA.

出版信息

Pathology. 2003 Dec;35(6):513-7. doi: 10.1080/00313020310001619910.

DOI:10.1080/00313020310001619910

PMID:14660102

Abstract

AIM

Renal cell carcinoma (RCC) represents the most common malignant tumour in the kidney and is resistant to conventional therapies. G250 is a tumour-associated antigen and is expressed mainly in clear cell RCC. It is a potential therapeutic target as well as a diagnostic marker for RCC. Previous studies relating to G250 protein or G250 mRNA expression in RCC using human tissue have largely been limited to immunohistochemical or conventional RT-PCR approaches that lack reproducibility, are subject to observer bias and are at most semi-quantitative. In this study, we evaluated the feasibility of using real-time RT-PCR in quantifying G250 mRNA expression using tissue dissected from paraffin sections of renal epithelial neoplasms. Using this approach, we performed a preliminary study investigating the relationship between G250 expression and tumor behavior in clear cell RCC.

METHODS

A total of 53 radical nephrectomy specimens with renal epithelial neoplasms (nine oncocytoma, seven chromophobe RCC, four papillary RCC and 33 clear cell RCC) were included in this study. SYBR Green real-time RT-PCR was performed using the ABI PRISM 7700 Sequence Detection System. A TATA box-binding protein (TBP) was used as an endogenous reference gene.

RESULTS

Our study showed that real-time RT-PCR is a reliable approach in quantifying G250 mRNA expression using paraffin sections. Using this methodology, G250 mRNA was detected in all of the clear cell RCCs, but none of the other types of tumours studied or in normal kidney tissue. Our study also suggested that the expression of G250 mRNA might correlate with tumour stage and overall survival.

CONCLUSION

Among common renal epithelial neoplasms, G250 is a specific marker for clear cell RCC. Quantification of G250 mRNA expression by real-time RT-PCR using paraffin sections is a feasible approach for future clinical studies in evaluating the prognostic and predictive value of G250 in clear cell RCC.

摘要

目的

肾细胞癌（RCC）是肾脏最常见的恶性肿瘤，对传统疗法具有抗性。G250是一种肿瘤相关抗原，主要在透明细胞RCC中表达。它是RCC的一个潜在治疗靶点及诊断标志物。以往使用人体组织对RCC中G250蛋白或G250 mRNA表达的研究，很大程度上限于免疫组织化学或传统逆转录聚合酶链反应（RT-PCR）方法，这些方法缺乏可重复性，存在观察者偏倚，且最多为半定量。在本研究中，我们评估了使用实时RT-PCR对从肾上皮肿瘤石蜡切片中解剖的组织进行G250 mRNA表达定量分析的可行性。采用这种方法，我们进行了一项初步研究，调查G250表达与透明细胞RCC肿瘤行为之间的关系。