食管癌细胞诱导裸鼠肝血管内皮细胞E-选择素的表达

Expression of E-selectin on Vascular Endothelial Cells in Nude Mice Liver Induced by Esophageal Carcinoma Cells

目的探讨食管癌细胞诱导裸鼠肝血管内皮细胞E-选择素(E-selectin)的表达,及其在食管癌肝转移中的作用。方法 Balb/c裸鼠12只,鼠龄6～8周,重量20～25g。将12只鼠随机分为对照组和实验组,每组6只。实验组鼠用食管癌细胞株EC9706悬液(5×106/0.02ml)注入脾包膜下,1h后切除脾;对照组同样行开腹手术,注射磷酸盐缓冲液(PBS)。注射8h后处死鼠取肝,用逆转录-聚合酶链式反应(RT-PCR)和免疫组织化学链霉亲和素生物素标记(LsAB)法检测裸鼠肝血管内皮细胞E-选择素的表达。结果实验组食管癌细胞株EC9706(5×106个)注射到裸鼠脾内,8h后用RT-PCR可在肝脏检测到E-选择素mRNA的表达;免疫组织化学检测显示:肝窦血管胞浆和胞膜E-选择素蛋白表达呈阳性。而对照组肝脏未能检测到E-选择素mRNA表达,E-选择素蛋白表达亦呈阴性。结论食管癌细胞株EC9706能诱导裸鼠肝脏E-选择素mRNA及蛋白表达,表明食管癌细胞株EC9706可能在肝脏中停留并形成转移灶。

Objective To study the expression of E-selectin on vascular endothelial cells of nude mice liver induced by esophageal carcinoma cells,in order to find out the function of E-selectin in the metastasis of esophageal carcinoma into the liver.Methods Twelve Balb/c nude mice aged from 6 to 8 weeks with their weight ranged between 20 and 25 grams were selected in our research.The mice were equally distributed into the experimental group and the control group（n=6）.EC9706 cell solution （5×106/0.02 ml） were injected beneath the splenic capsule of the mice in the experimental group.One hour later,spleen was removed.For the mice in the control group,after laparotomy,phosphate buffer without EC 9706 was injected beneath the splenic capsule and spleen was also removed one hour after the injection.Eight hour later,we resected the liver of the nude mice,and expression of E-selectin on vascular endothelial cells of the liver was detected with reverse transcription polymerase chain reaction （RT-PCR） and immunohistochemistry （IHC）.Results In the experimental group,8 hours after injection of EC9706 cells （5×106）,the results of RT-PCR showed expression of E-selectin mRNA in the liver,and IHC showed a positive protein expression of E-selectin in the cytosol and membrane of hepatic sinus vessels.However,no E-selectin mRNA expression was found in the control group and IHC showed a negative protein expression of E-selectin.Conclusion Human esophageal carcinoma cell line EC9706 can induce balb/c mice liver vascular endothelial cell E-selectin expression,which shows that EC9706 may stay in the liver and form metastatic focus.