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Quantitative Proteomics Analysis of Differentially Expressed Proteins Involved in Renal Clear Cell Carcinoma by Shotgun Approach Coupled with Stable Isotope Dimethyl Labeling

Shih-Shin Liang, Chao-Jen Kuo, Shu-Wen Chi, Wen-Jeng Wu, Shui-Tein Chen and Shyh-Horng Chiou

Renal clear cell carcinoma (RCC) when detected incidentally by ultrasonic imaging generally shows an advanced disease stage of being diagnosed metastatic as a result of lacking specific biomarkers at an early stage. To date, RCC is still ranked as the most common renal cancer and considered as one of the most treatment-resistant metastatic malignancies. In this study, a quantitative proteomics approach by means of gel-free shotgun proteomics methodology and stable isotope dimethyl labeling coupled with nano-liquid chromatography/ tandem mass (nanoLCMS/ MS) have been employed to identify 18 up-regulated and 48 down-regulated proteins in RCC samples. It is worth noting that binding and structural proteins in renal clear cells accounted for 43% and 33% of up-regulated proteins respectively, while catalytic enzymes occupied as high as 73% of down-regulated proteins. Collectively, instead of one universal tumorigenesis enzyme being identified, the pathogenesis of RCC may involve a variety of protein factors including ANXA2, LGALS1, VIM and TPM1 related to metastasis, angiogenesis, tumor invasion as well as tumor growth, in addition to CRYAB, GSTA1, CALB1 and HSPD1, which are linked to apoptosis function. The upshot of this study highlighted by a series of down-regulated proteins suggests that the clear-cut decrease of ATP generation components related to mitochondrial dysfunction and altered energy metabolism may be involved in RCC carcinogenesis.