Panel of candidate biomarkers for renal cell carcinoma.
The timely diagnosis and therapeutic monitoring of human renal cell carcinoma (RCC) is limited by the lack of specific biomarkers. To identify candidate RCC biomarkers, we used 2-DE gel electrophoresis with mass spectrometry and 2-DE spot intensity-based ROC analysis to analyze 18 sets of paired normal and RCC tumor tissue including conventional, papillary, and chromophobe subtypes. Validation was performed with RCC patient plasma samples and confirmed by clustergram, shRNA, and immunohistochemistry assays. Cardinal candidates were evaluated by ELISA. The leading candidate biomarker that was upregulated in RCC samples according to the clustergram and validation analysis was nicotinamide N-methyltransferase (NNMT) (13/15, P < 0.0001). Other upregulated candidate biomarkers that were identified by this method include ferritin, hNSE, NM23, secretagogin, and L-plastin. The upregulation of NNMT in RCC was confirmed by immunoblotting and immunohistochemistry. Analysis of fractionated membrane-associated proteins identified CAP-G, mitofillin, tubulin alpha, RBBP7, and HSP27. Of these, RBBP7 and HSP27 were highly expressed in the chromophobe subtype of RCC (3/3) but were absent from conventional RCC (0/3). The triple combination of the NNMT, FTL, and hNSE biomarkers had the highest predictive capacity of 0.993, while NNMT was the single, most powerful candidate diagnostic biomarker for all types of RCC.
Kim
Dong Su
DS
Research Division, DCD, Inc., Pohang Technopark, Pohang, Kyungbuk, 790-834, Korea.
Choi
Yoon Pyo
YP
Kang
Suki
S
Gao
Ming Qing
MQ
Kim
Baekil
B
Park
Haeng Ran
HR
Choi
Young Deuk
YD
Lim
Jong Baek
JB
Na
Hyung Jin
HJ
Kim
Hye Kyung
HK
Nam
Young-Pyo
YP
Moon
Mi Hyang
MH
Yun
Hae Ree
HR
Lee
Dong Hee
DH
Park
Won-Man
WM
Cho
Nam Hoon
NH
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000032:analysis / Q000378:metabolism*; D002292:Carcinoma, Renal Cell / Q000175:diagnosis / Q000378:metabolism*; D045744:Cell Line, Tumor; D016000:Cluster Analysis; D015536:Down-Regulation; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D007680:Kidney Neoplasms / Q000175:diagnosis / Q000378:metabolism*; D008297:Male; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism; D008875:Middle Aged; D050903:Nicotinamide N-Methyltransferase / Q000032:analysis / Q000378:metabolism*; D020543:Proteome / Q000032:analysis / Q000378:metabolism*; D012372:ROC Curve; D015203:Reproducibility of Results; D015854:Up-Regulation
D014408:Biomarkers, Tumor; D008565:Membrane Proteins; D020543:Proteome; C494406:NNMT protein, human; D050903:Nicotinamide N-Methyltransferase