Objective This study aimed to analyze the differences between the human placenta cancer cell line JAR and the methotrexateresistant JAR / MTX and screen out the drug resistance-associated proteins in the choriocarcinoma. Methods The protein was extracted from culture cell and to examine the cytoplasmic and secreted proteins using the surface-enhanced laser desorption ionization-time of flight mass spectrometry (SELDI-TOF- MS). This cell lines were tested using CM-10 weak cation exchanging protein chip. Results The cytosolic proteins with mass charge ratio(m/z) of 8 132.043,10 738.36 in the cytoplasmic protein was highly expressed in JAR / MTX cells. The secreted protein with mass charge ratio (m/z) of 11 555.09 and the cytolosic protein with the mass charge ratio (m/z) of the 4 896.68 was highly expressed in JAR cells, but lowly in JAR/MTX cells. Conclusion The combination of the time of flight mass spectrometry technology with a weak cation exchanging protein chip might effectively screen specific proteins of MTX-resistant choriocarcinoma cell line and explore the relation between protein expression and drug resistance in the choriocarcinoma.%目的 应用飞行时间质谱技术结合蛋白芯片分析筛选人绒毛癌细胞株JAR及其与人绒毛癌耐氨甲喋呤细胞株JAR/MTX之间的蛋白质表达差异,寻找出绒癌耐药的相关蛋白.方法 采用体外细胞培养来提取两种细胞的胞浆蛋白和分泌蛋白,应用SELDI-TOF-MS技术检测各细胞株提取的胞浆蛋白和分泌蛋白,采用CM-10型弱阳离子交换芯片检测.结果 经SELDI技术检测发现质荷比(m/z)为8132.043,10738.36的胞浆蛋白在JAR/MTX细胞中呈高表达;质荷比(m/z)为11555.09的分泌蛋白及m/z为4896.68的胞浆蛋白在JAR细胞中高表达,在JAR/MTX中呈低表达.结论 应用飞行时间质谱技术结合弱阳离子交换芯片,可有效筛选绒癌耐氨甲喋呤细胞株中的特异性表达蛋白;寻找到的差异蛋白与绒癌耐药密切相关,并可能成为本病耐药的标志物.
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