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Quantitative Proteomics Profiling of RKO and HT29 Cell Lines by iTRAQ Coupled LC-MS/MS

[ Vol. 15 , Issue. 3 ]

Author(s):

Jie Zhao*, Yuhong Wang, Xiaoyuan Jia and Xueqing Yao   Pages 241 - 256 ( 16 )

Abstract:


Background: Metastasis often happened in colorectal cancer patients. It is still unclear on the metastatic mechanism on colorectal cancer. Hence, we used a proteomic method to find the multiplex markers associated with metastasis. The colorectal cancer cell lines RKO and HT29 had been identified as the hepatic metastasis model in vitro.

Method: Here, we compared the two cell proteome profiles and found some new molecular events of CRC metastasis. Using iTRAQ based proteomics method, we identified 1985 specific proteins, out of which 212 were found to be significantly changed in two cell lines. 28 candidate genes were confirmed by Q-PCR and 9 candidate proteins were further validated by western blot.

Results: Bioinformatics analysis revealed the altered proteins played roles in actin cytoskeleton, focal adhesion and so on. KEGG analysis of the altered proteins indicated several signal pathways related to tumor invasion and metastasis. The expression of some genes was related to epigenetic changes. These results may supply an perspicacity with some new molecular biomarkers development in colorectal cancer metastasis.

Conclusion: In a word, this study had identify several novel moleculars for CRC metastasis and development.

Keywords:

Colorectal cancer, metastasis, iTRAQ, proteomics, marker, ladinin 1.

Affiliation:

School of Medicine, South China University of Technology, Guang Zhou 510006, School of Medicine, Zhejiang University, Hang Zhou 310012, College of Life Science, Zhejiang Sci-Tech University, Hang Zhou 310018, School of Medicine, South China University of Technology, Guang Zhou 510006

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