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Originally published In Press as doi:10.1074/mcp.M500055-MCP200 on June 20, 2005.
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Molecular & Cellular Proteomics 4:1480-1486, 2005.
© 2005 by The American Society for Biochemistry and Molecular Biology, Inc.

Research

An Approach to Studying Lung Cancer-related Proteins in Human Blood*,S

Ting Xiaoa,b, Wantao Yingc,b, Lei Lic,b, Zhi Hua,b, Ying Maa, Liyan Jiaoc, Jinfang Maa, Yun Caic, Dongmei Lind, Suping Guoa, Naijun Hana, Xuebing Dia, Min Lia, Dechao Zhange, Kai Sue, Jinsong Yuana, Hongwei Zhenga, Meixia Gaoa, Jie Hea, Susheng Shid, Wuju Lif, Ningzhi Xua, Husheng Zhanga, Yan Liua, Kaitai Zhangc,g, Yanning Gaoa,h, Xiaohong Qianc,i and Shujun Chenga,j

From the Departments of a Etiology and Carcinogenesis, d Pathology, and e Chest Surgery, Cancer Institute (Hospital), Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100021, China, c Department of Genomics and Proteomics, Beijing Institute of Radiation Medicine and f Center of Computational Biology, Beijing Institute of Basic Medical Sciences, 27 Taiping Road, Beijing 100850, China

Early stage lung cancer detection is the first step toward successful clinical therapy and increased patient survival. Clinicians monitor cancer progression by profiling tumor cell proteins in the blood plasma of afflicted patients. Blood plasma, however, is a difficult cancer protein assessment medium because it is rich in albumins and heterogeneous protein species. We report herein a method to detect the proteins released into the circulatory system by tumor cells. Initially we analyzed the protein components in the conditioned medium (CM) of lung cancer primary cell or organ cultures and in the adjacent normal bronchus using one-dimensional PAGE and nano-ESI-MS/MS. We identified 299 proteins involved in key cellular process such as cell growth, organogenesis, and signal transduction. We selected 13 interesting proteins from this list and analyzed them in 628 blood plasma samples using ELISA. We detected 11 of these 13 proteins in the plasma of lung cancer patients and non-patient controls. Our results showed that plasma matrix metalloproteinase 1 levels were elevated significantly in late stage lung cancer patients and that the plasma levels of 14-3-3 {sigma}, ß, and {eta} in the lung cancer patients were significantly lower than those in the control subjects. To our knowledge, this is the first time that fascin, ezrin, CD98, annexin A4, 14-3-3 {sigma}, 14-3-3 ß, and 14-3-3 {eta} proteins have been detected in human plasma by ELISA. The preliminary results showed that a combination of CD98, fascin, polymeric immunoglobulin receptor/secretory component and 14-3-3 {eta} had a higher sensitivity and specificity than any single marker. In conclusion, we report a method to detect proteins released into blood by lung cancer. This pilot approach may lead to the identification of novel protein markers in blood and provide a new method of identifying tumor biomarker profiles for guiding both early detection and therapy of human cancer.

g To whom correspondence may be addressed. E-mail: zhangkt{at}vip.sina.com

h To whom correspondence may be addressed. E-mail: yngao{at}pubem.cicams.ac.cn

i To whom correspondence may be addressed. Tel.: 86-10-68279585; Fax: 86-10-68279585; E-mail: qianxh{at}nic.bmi.ac.cn

j To whom correspondence may be addressed: P O. Box 2258, Beijing 100021, China. Tel.: 86-10-6776-6703; Fax: 86-10-6776-7548; E-mail: chengshj{at}263.net.cn


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