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


CONCLUSIONS
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.
Bismar
Tarek A
TA
Department of Pathology, Harper University Hospital, Wayne State University, 3990 John R., Detroit, MI 48201, USA.
Bianco
Fernando J
FJ
Zhang
Hongquan
H
Li
Xingli
X
Sarkar
Fazlul H
FH
Sakr
Wael A
WA
Grignon
David J
DJ
Che
Mingxin
M
D000951:Antigens, Neoplasm / Q000235:genetics*; D014408:Biomarkers, Tumor / Q000032:analysis*; D000071231:Carbonic Anhydrase IX; D002256:Carbonic Anhydrases / Q000235:genetics*; D006801:Humans; D007680:Kidney Neoplasms / Q000235:genetics*; D009363:Neoplasm Proteins / Q000235:genetics*; D009375:Neoplasms, Glandular and Epithelial / Q000235:genetics*; D010232:Paraffin; D011379:Prognosis; D012333:RNA, Messenger / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D010232:Paraffin; C089226:CA9 protein, human; D000071231:Carbonic Anhydrase IX; D002256:Carbonic Anhydrases