应用SELDI-TOF-MS技术筛选卵巢癌淋巴结定向高转移特性细胞株差异表达蛋白  

Screening of Differentially Expressed Proteins Associated with Directional Highly Lymphatic Metastasis in Ovarian Carcinoma Cell Lines Using SELDI-TOF-MS Technology

作　　者：冠潇[1] 黎丹戎[1] 王琪[1] 李力[1] 阮和云[1] 张玮[1] 

机构地区：[1]广西医科大学附属肿瘤医院妇瘤科,广西南宁530021

出　　处：《癌症》2008年第11期1155-1160,共6页Chinese Journal of Cancer

基　　金：教育部高等学校博士学科点专项科研基金项目(20050598002);广西自然科学基金项目(桂科自0728193;桂科能0630006-5E2Z);广西自治区大型仪器协作共用网资助项目~~

摘　　要：背景与目的:人卵巢浆液性乳头状腺癌细胞株SKOV3及其在裸鼠体内建立的淋巴结定向高转移亚克隆第二代SKOV3-pm2、第三代SKOV3-pm3细胞株,为卵巢癌转移分子机制的研究提供了细胞模型。本研究应用飞行时间质谱技术结合蛋白芯片分析筛选卵巢癌淋巴结定向高转移与非定向转移细胞株之间的蛋白质表达差异。方法:采用动物实验测定SKOV3、SKOV3-pm2和SKOV3-pm3三种细胞株的淋巴结转移率。应用表面增强激光解吸离子化-飞行时间质谱(surface-enhanced laser desorption and ionization-time of flight-mass spectrometry,SELDI-TOF-MS)技术检测各细胞株提取的胞浆总蛋白和分泌蛋白,每种细胞分别采用CM-10型弱阳离子交换芯片和IMAC-3型固定金属亲和芯片检测。应用Ciphergen Protein Software3.2.0软件和Biomarker Wizard软件对采集3组细胞的蛋白峰进行比较,峰强度相差0.5倍以上者定义为差异蛋白。结果:动物实验显示,细胞株SKOV3及高转移亚克隆SKOV3-pm2、SKOV3-pm3的淋巴结转移率分别为20%、90%和100%,前者与后二者之间的差异有统计学意义(P<0.05)。获得CM-10和IMAC3两种蛋白芯片上SKOV3、SKOV3-pm2、SKOV3-pm3细胞株蛋白质谱图谱,对比发现:质荷比(m/z)为6971、7475、9089、9453、10103、11655的胞浆蛋白及质荷比(m/z)为4746的分泌蛋白在上述三种细胞株中存在不同程度差异表达。结论:应用SELDI-TOF-MS技术结合固定金属亲和芯片和弱阳离子交换芯片,可有效筛选卵巢癌淋巴结定向高转移特性细胞株中的特异性表达蛋白;寻找到的差异蛋白与淋巴结定向高转移潜能密切相关。BACKGROUND ＆ OBJECTIVE. The ovarian serous papillary adenocarcinoma cell line SKOV3 and its subclones SKOV3-pm2 and SKOV3- pro3 are cell models to investigate the molecular mechanism of invasion and metastasis of ovarian cancers. This study was to screen differentially expressed proteins between ovarian carcinoma cell lines with directional （SKOV3-pm2 and SKOV3-pm3） and non-directional （SKOV3） highly lymphatic metastasis potentials using time-of-flight mass spectrometry technology and protein chips. METHODS： The lymphatic metastasis rates of the three cell lines were detected in animal models. Proteins in endochylema and supernatants of the three cell lines were screened using surface- enhanced laser desorption and ionization-time of flight-mass spectrometry （SELDI-TOF-MS）. Each sample was examined using weak cation exchange （CM-10） protein chip assay and immobilized metal affinity capture （IMAC-3） SELDI ProteinChip array. Detected protein peaks were filtrated and analyzed using Ciphergen proteinchip software 3.2.0 and Biomarker Wizard software. Differentially expressed proteins were defined as those whose absolute ratio values were greater than 0.5. RESULTS：Lymphatic metastasis rates in SKOV3, SKOV3-pm2 and SKOV3-pm3 cell xenografts in nude mice were 20%, 90% and 100%, respectively （P〈0.05）. Proteins in endochylema whose mass-to-charge ratio （m/z） were 6971, 7475, 9089, 9453, 10103, 11655, and the protein in supernatants whose m/z was 4746 were differentially expressed in SKOV3, SKOV3-pm2 and SKOV3-pm3 cells. CONCLUSION： Combined with weak cation exchange protein chip assay and immobilized metal affinity capture SELDI ProteinChip array, SELDI-TOFMS technology can be used to screen and identify differentially expressed proteins associated with directional highly lymphatic metastasis in ovarian carcinoma cell lines.

关 键 词：卵巢肿瘤 SKOV3细胞株 SKOV3一pm2细胞株 SKOV3-pm3细胞株 蛋白芯片 差异蛋白 表面增强激光 解吸离子化:飞行时间质谱 

分 类 号：R737.31[医药卫生—肿瘤]