來自中科院北京基因組研究所蛋白質(zhì)組學(xué)組,，中國醫(yī)學(xué)科學(xué)院& 中國協(xié)和醫(yī)科大學(xué)腫瘤研究所（腫瘤醫(yī)院）等處的研究人員通過對一種M-BE細胞系進行蛋白質(zhì)組學(xué)分析，發(fā)現(xiàn)了一種可以作為潛在肺癌早期診斷的生物標(biāo)記物,，為肺癌治療與檢測提出了一種新方法,。這一研究成果公布在《J. Proteome Res.》雜志上,。
領(lǐng)導(dǎo)這一研究的是來自中國醫(yī)學(xué)科學(xué)院的徐寧志教授，其早年畢業(yè)于中國醫(yī)學(xué)科學(xué)院腫瘤研究所,，主要研究領(lǐng)域包括從細胞,、染色體、基因,、蛋白質(zhì)等不同層次與水平研究腫瘤的發(fā)生機制；研究細胞癌變的分子機制和信號傳導(dǎo)通路,，以及細胞增殖,、凋亡與惡性轉(zhuǎn)化的信號傳導(dǎo)通路等。
肺癌是最常見的肺原發(fā)性惡性腫瘤,，絕大多數(shù)肺癌起源于支氣管粘膜上皮,，故亦稱支氣管肺癌。近50多年來,，世界各國特別是工業(yè)發(fā)達國家,，肺癌的發(fā)病率和病死率均迅速上升,，死于癌病的男性病人中肺癌已居首位。
肺癌的早期診斷是減少惡性腫瘤致死的一種有效方法,，有假說認為肺癌的臨床證據(jù)是原癌基因或腫瘤抑制基因先天或后天逐漸發(fā)生的不同變異的積累,，該過程產(chǎn)生了肺癌標(biāo)記物。許多標(biāo)記物可以反映腫瘤大小,、癌細胞溶解的速度和腫瘤其他潛在的惡性特質(zhì),，可作為肺癌評價和臨床進展演變監(jiān)測的易行而有效的途徑。
在這篇文章中,，研究人員為了尋找肺癌早期血清生物標(biāo)記物,，將一種SV40T-transformed人類支氣管上皮細胞系（human bronchial epithelial cell line）：M-BE放置于條件培養(yǎng)基中培育，檢測其分泌蛋白,。研究人員首先將不同傳代的M-BE細胞分泌出來的蛋白提取出來,，利用2-DE進行分離，并通過MALDI-TOF/TOF質(zhì)譜分析識別2-DE,。
這樣研究人員一共識別了47個蛋白,，其中23個屬于正調(diào)控蛋白，24個負調(diào)控蛋白,。在這些蛋白中,，cathepsin D被認為是一種能在培養(yǎng)基和細胞中豐度遞增的典型分泌蛋白，而且在肺癌患者臨床樣品中也得到了蛋白質(zhì)組學(xué)結(jié)論,。之后利用三明治ELISA實驗,，血漿中cathepsin D的濃度在肺部squamous cell carcinomas (SCC, 104 例) 正常細胞 (36 cases, p 0.015)中呈現(xiàn)出暨大區(qū)別。
這說明不同傳代M-BE細胞可以分泌或釋放一些蛋白到環(huán)境中,，其中cathepsin D也許能作為一種肺癌生物標(biāo)記物的潛在來源,，為肺癌早期檢測提供新的方法。
原始出處：
J. Proteome Res., 6 (3), 1083 -1092, 2007. 10.1021/pr060422t S1535-3893(06)00422-2
Web Release Date: February 7, 2007 Copyright © 2007 American Chemical Society
Cathepsin D Is Secreted from M-BE Cells: Its Potential Role as a Biomarker of Lung Cancer
Xiaomin Lou,# Ting Xiao,# Kang Zhao, Hao Wang, Hongwei Zheng, Dongmei Lin, Youyong Lu, Yanning Gao, Shujun Cheng, Siqi Liu, and Ningzhi Xu*
Division of Proteomics, Beijing Genomics Institute, Chinese Academy of Sciences, Beijing 101318, P. R. China, Department of Etiology and Carcinogenesis, Cancer Institute & Cancer Hospital, Chinese Academy of Medical Sciences & Peking union Medical College, Beijing 100021, P. R. China, Peking University School of Oncology, Beijing Institute for Cancer Research, Beijing 100034, P. R. China, Laboratory of Cell and Molecular Biology, Cancer Institute & Cancer Hospital, Chinese Academy of Medical Sciences & Peking union Medical College, Beijing 100021, P. R. China, and Graduate School of Chinese Academy of Sciences, Beijing 100049, P. R. China
Received August 20, 2006
Abstract:
The early diagnosis of lung cancer is an effective approach to reduce the mortality caused by malignancy. To explore serum biomarkers of lung cancer at early stage, M-BE, a SV40T-transformed human bronchial epithelial cell line with the phenotypic features of early tumorigenesis at high passage, was cultured in the conditioned media to collect its secretory proteins. The proteins secreted from different passage M-BE cells were extracted and then separated by two-dimensional electrophoresis (2-DE). MALDI-TOF/TOF mass spectrometry was adopted to identify the passage-dependent 2-DE spots. Totally, 47 proteins were identified, including 23 that were up-regulated and 24 that were down-regulated. Of these proteins, cathepsin D was a typical secretory protein that exhibited the increased abundance either in culture media or in cells during passaging. Furthermore, the proteomic conclusions were validated in the clinical samples of lung cancer patients. When sandwich ELISA was used, the concentrations of cathepsin D in plasma showed significant differences between lung squamous cell carcinomas (SCC, 104 cases) and normal donors (36 cases, p 0.015). When tissue microarray (TMA) was used, cathepsin D expression levels in SCC tissues (178 cases) were significantly higher than those in normal donors (40 cases, p < 0.001). The present study has revealed that M-BE cells at different passages could secrete or release some proteins into the living environment, which might serve as the potential resource for exploring the biomarkers of lung cancer.
Keywords: lung cancer conditioned media secretory protein Cathepsin D
附：
徐寧志
1986年畢業(yè)于中國醫(yī)學(xué)科學(xué)院腫瘤研究所,，獲醫(yī)學(xué)碩士學(xué)位,，
1989年－1996年在美國國家衛(wèi)生研究院（NIH），從事細胞癌變的分子生物學(xué)機理和信號傳導(dǎo)通路等研究,。
1997年3月回到中國醫(yī)學(xué)科學(xué)院腫瘤研究所工作,，任細胞生物學(xué)與分子生物學(xué)研究室主任，博士生導(dǎo)師,。
國家自然科學(xué)基金生命科學(xué)部學(xué)科評審組專家,，北京抗癌協(xié)會理事。主要學(xué)術(shù)研究方向是從多個水平研究惡性腫瘤發(fā)病的分子機理,，尤其是信號傳導(dǎo)通路異常在癌變過程中的作用,。主持973項目、國家杰出青年基金,、國家自然科學(xué)基金,，教育部博士點基金等多個基金項目,，已培養(yǎng)研究生6名，在讀5名,。共發(fā)表學(xué)術(shù)論文40余篇,。參編英文專著兩部，以及《腫瘤學(xué)》,、《臨床腫瘤學(xué)》等中文專著六部,。
多次主持或參加國家重大科研項目，現(xiàn)主要研究方向是從細胞,、染色體,、基因、蛋白質(zhì)等不同層次與水平研究腫瘤的發(fā)生機制,；研究細胞癌變的分子機制和信號傳導(dǎo)通路,，以及細胞增殖、凋亡與惡性轉(zhuǎn)化的信號傳導(dǎo)通路,。
e-mail: [email protected]
