摘要
目的探讨T—Synthase活性对胃癌组织中Tn、STn、T、ST抗原表达的影响。方法不连续密度梯度离心分离胃癌组织中的肿瘤细胞并进行培养，流式细胞仪检测胃癌组织中Tn＋与Tn-细胞的表达状况；免疫磁珠分选Tn＋、Tn-细胞，Westernblot检测Tn、STn、T、ST抗原的表达状况，RT—PCR检测T—SynthasemRNA的表达水平，液体闪烁计数仪检测T—Synthase活性。结果39．66％的病人胃癌细胞均为Tn＋细胞，29．31％Tn＋、Tn-I细胞同时存在，31．03％均为Tn-细胞；Tn＋细胞同时表达Tn／STn抗原，不表达T／sT抗原，Tn＋I及Tn-细胞均表达sT抗原，部分Tn。I细胞可见T抗原表达；Tn＋、Tn- I及Tn-细胞间T—SynthasemRNA无明显差别，T．Synthase活性在Tn’细胞明显下降。结论不同TNM期胃癌组织中Tn、STn、T、ST抗原表达存在差异，T．Synthase活性降低可能导致胃癌细胞表达Tn抗原。
Objective To explore the effects of T-Synthase activity on the expression of tumor-as- sociated carbohydrate antigens（TACAs）, Tn, STn, T and ST in gastric carcinoma tissues. Methods The tumor cells were separated from gastric carcinoma tissues of different TNM stages by density gradient centrifu- gation, and the counts of Tn ~ and Tn- cells were further tested by flow cytometry （FCM）. The Tn ＋ and Tn- cells were selected from cultured gastric tumor cells by immune magnetic bead. In these cells the expression of Tn, STn, T and ST antigen in membrane extracts, and the T-Synthase mRNA were detected by Western blot and RT-PCR respectively, then using GalNAcal-O-phenyl as acceptor for UDP-[ H3]- Gal, T-Synthase activity in cell extracts were determined by radioactivity. The relationship between Tn antigen and T-Synthase activity were analyzed. Results 39.66% patients＇ gastric tumor cells are only Tn ~ cells, 29.31% pa- tients＇ cells are mixture of Tn ~ and Tn- I ceils, 31.03% patients＇ cells are only Tn- cells. Tn and STn anti- gen co-expressed in Tn ＋ cells, have no T and ST antigen. ST antigen expressed in both of Tn- I and Tn- cells, and T antigen expressed in proportion of Tn- I cells. No difference of T-Synthase mRNA was detected between Tn ~ , Tn I and Tn- cells, but T-Synthase activity in Tn ~ cell extracts was much lower than that in Tn- cell. Conclusion The expression of Tn, STn, T and ST antigen in gastric carcinoma tissues of different TNM stages is different. Tn antigen expression in tumor cells may be caused by the decrease of T-Synthase activity.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2009年第8期762-766,共5页
Chinese Journal of Microbiology and Immunology
基金
基金项目：山东省自然科学基金（Y2007C143）
