应用SELDI-TOF-MS技术筛选人肝癌细胞多药耐药相关差异表达蛋白

Screen the differential expressed proteins as to multidrug resistance in human hepatocellular carcinoma cell line by SELDI-TOF-MS

【目的】通过SELDI-TOF-MS技术筛选体外培养人肝癌细胞多药耐药相关差异表达蛋白。【方法】采用体外培养人肝癌细胞Bel-7402及其耐5-氟尿嘧啶细胞Bel-FU,MTT法检测Bel-FU的多药耐药性;SELDI-TOF-MS技术结合CM-10芯片检测Bel-7402与Bel-FU蛋白质指纹图谱。【结果】细胞Bel-FU对于5-氟尿嘧啶、阿糖胞苷、长春新碱、阿霉素、柔红霉素的耐药指数分别为:89.6倍、3.9倍、37.2倍、16.5倍、3.7倍。在质荷比(M/Z)2 000～50 000的范围内,Bel-FU与Bel-7402间共检测到8个差异蛋白(P<0.05),其中上调蛋白有3个,下调蛋白有5个。【结论】SELDI-TOF-MS技术为筛选肝癌MDR蛋白标志物提供了一个简便、有效的方法。

【Objective】 To sreen the differential expressed proteins as to multidrug resistance in human hepatocellular carcinoma cell lines cultured in vitro by SELDI-TOF-MS.【Methods】 Human hepatocellular carcinoma cell lines BEL-7402 and multidrug-resistant hepatocellular carcinoma cell lines Bel-Fu were cultured in vitro.The multidrug resistance（MDR） of Bel-Fu was detected by methyl thiazolyl tetrazolium（MTT） assay.Protein fingerprints about Bel-7402 and Bel-FU was detected by SELDI-TOF-MS and CM-10 chips.【Results】 The resistance index（IR） of Bel-FU for 5-fluorouracil,cytarabine,vincristin,doxorubicin,daunorubicin were 89.6,3.9,37.2,16.5 and 3.7 respectively.Ranging from 2 000~50 000（M/Z）,8 differential expressed proteins were detected between cell lines Bel-7402 and Bel-Fu（P0.05）,3 differential expressed proteins were up-regulated and 5 differential expressed proteins were down-regulated.【Conclusion】 SELDI-TOF-MS provides a convenient and effective method for screening biomarkers of HCC Multidrug Resistance.