人胃肠道间质瘤细胞系的建立及其永生化

Establishment and Immortalization of Human Gastrointestinal Stromal Tumor Cells

目的:建立体外培养的永生化胃肠道间质瘤细胞系,并测定其对STI571的药物敏感性。方法:用中性蛋白酶消化法从胃肠道间质瘤新鲜标本中分离出间质瘤细胞,以含端粒酶逆转录酶(hTERT)和猿猴病毒40大T抗原(SV40LT)的重组逆转录病毒感染对其进行永生化;然后,用MTT法测定STI571对该细胞系的药物敏感性。结果:永生化的胃肠道间质瘤细胞,呈短梭形或不规则形状,单层生长。Western blot检测表明该细胞系表达c-kit、c-abl、hTERT、SV40LT蛋白。MTT结果显示STI571对此细胞有较显著的抑制作用。结论:建立的细胞系具有胃肠道间质瘤细胞的特征,可作为体外研究胃肠道间质瘤发病和耐药机制的重要手段之一。

Objective：To establish a grastrointestinal stromal tumor（GIST） cell line and to determine its drug sensitivity to STI571.Methods：GIST cells were separated from fresh GIST specimens through dispase digestion and then were immotalized by ectopic expression of telomerase reverse transcriptase（hTERT） and simian virus 40 large T（SV40LT） antigen.The effect of STI571 on this cell line was detected by MTT dye.Results：The cells showed fusiform shape and formed monolayer in vitro culture;they expressed c-kit,hTERT,and SV40LT antigens.STI571 significantly inhibited the proliferation of the cells.Conclusion：The cells have the properties of GIST and can be used as an important tool for studying the carcinogenesis and resistant mechanism of GIST.