WtCosmc质粒转染对Tn^+肿瘤细胞恶性行为的影响

Study on the malignant behavior of Tn^+ tumor cells after transcription with wild type Cosmc

目的：以Tn＋肿瘤细胞为靶标,研究野生型Cosmc（WtCosmc）质粒转染对其恶性行为的影响,以期为肿瘤的诊断及治疗提供新思路.方法：流式细胞术检测肿瘤细胞表面Tn抗原阳性率,免疫磁珠分选Tn＋与Tn-肿瘤细胞;分选后的Tn＋肿瘤细胞经Fugene 6转染WtCosmc质粒,并用免疫荧光检测细胞表面Tn抗原表达状况.分别采用荧光法、CCK-8和Transwell检测转染前后Tn＋肿瘤细胞的T-synthase活性、细胞增殖、迁移能力及对Apo2L/TRAIL的敏感性.结果：与Tn-细胞相比,Tn＋肿瘤细胞T-synthase活性及对Apo2L/TRAIL诱导凋亡的敏感性较低;细胞增殖、迁移能力较强.经WtCosmc质粒转染后,Tn＋细胞T-synthase活性及其对Apo2L/TRAIL诱导凋亡的敏感性明显增高;Tn抗原表达受抑;细胞增殖及迁移能力明显下降.结论：转染WtCosmc可恢复Tn＋细胞T-synthse活性,抑制Tn抗原的表达,进而增强其对Apo2L/TRAIL诱导凋亡的敏感性,降低细胞增殖、迁移能力,有效抑制Tn＋肿瘤细胞的恶性行为.

Objective In order to provide a new idea of diagnosis and treatment for human tumor% Tn positive（Tn＋ ） tumorcells were regard as targets to stuJy its change of malignant behavior affected by WtCosmc transfection. Methods： The percentage of Tn antigen expression in tumor cells line was detected by flow cytometry（FCM） % Tn antigen positive（Tn＋ ） and negative （Tn ） tumor cells in theirs were separated by immune magnetic bead. T-synthase activity in by fluorescent assay using GalNAc-cr（4-MU） as acceptor and UDP-Gal as the donor. The status of Tn antigen expression in Tn＇ cells transfected by wild-type （WtCosmc） using Fugene 6 transfection reagent were examined through immunofluorescence. Before and after being treated with WtCosmc% the ability of tumor cell proliferation monitored with the Cell Counting Kit 8 （CCK8） % and the ability of tumor cell migrationResults： Compared with Tn cells% T-synthase activity and the sensitivity of tumor lower in Tn＇ tumor cells% while the ability of migration and proliferation was much higher. After transfected with WtCosmc%T-synthase activity was restored and Tn antigen expression was decreased in Tn＇ tumor cells. Moreover % the ability ofproliferation, metastasis of Tn＇ cells was reduced% and these cells were more sensitive to Apo2LWtCosmc transfection could restore T-synthase activity and eliminate expression of the Tn antigens in Tn＇ tumor cells.Furthermore% Tn＇ tumor cells were more sensitive to Apo2L/TRAIL after WtCosmc transfection% which contributes toinhibit its malignant behavior.