LCM、IMB联合MALDI-TOF-MS技术在恶性卵巢上皮瘤相关标志物分离检测中的应用

摘要

目的 排除混杂细胞的干扰,获取人卵巢上皮瘤细胞,分离、检测特异相关标志物.方法 采用激光捕获显微切割 (LCM) 技术获取无间质混杂的人正常、良性、恶性卵巢上皮瘤细胞,分别提取细胞总蛋白,用免疫磁珠(IMB)技术和基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)技术分离检测黑色素瘤激活因子CXCL1及IL-8 (1～77 aa)、IL-8(6～77 aa)和IL-8(9～77 aa).结果 利用LCM获取的卵巢上皮细胞数量级达106～107.IMB可将CXCL1及不同亚型IL-8从卵巢正常、良性和恶性上皮瘤细胞的蛋白裂解液中分离并用于MALDI-TOF-MS检测.结论 LCM、IMB联合MALDI-TOF-MS技术可以准确有效地获取卵巢上皮癌细胞的特异标志物.%Objective To exclude the disturb of mixed cell types, segregate and detecte specific biomarkers of human ovarian tumor. Methods Pure cell from normal, nonmalignant and malignant ovarian tissues were captured by the laser capture microdissection(LCM). After cell protein isolated,the specific biomarkers of human ovarian tumor( the different subtype IL-8 and the melanoma activating factor CXCL1 ) were gathered and detected by the immunomagnetic beads(IMB)and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) respectively. Results The pure epithelial cells from ovrian tissues were well discriminated, which get to the 106 ～ 107 order of magnitude. The specific biomarkers IL-8(1 ～77 aa), IL-8(6 ～77 aa), IL-8(9 ～77 aa) and the CXCL1 could be admirably gathered by the IMB and be further detected by the MALDI-TOF-MS. Conclusion The technology platform of LCM and IMB coupled with MALDI-TOF-MS could correctly acquire the human ovarian tumor related biomarkers.