癌胚抗原过表达对结肠癌细胞生物学特性的影响

Effect of carcinoembryonic antigen overexpression on biological characteristics of colon cancer cells

目的观察癌胚抗原（CEA）过表达对结肠癌细胞生物学特性的影响。方法以重组CEACAM5一eGFP—cDNA慢病毒和荧光对照增强型绿色荧光蛋白（eGFP）慢病毒分别转染鼠结肠癌CT26细胞，构建稳定表达癌胚抗原的CT26CEA单克隆细胞株和仅表达荧光蛋白的CT26GFP单克隆细胞株，并以逆转录一聚合酶链反应（RT—PCR）和Westernblot法进行鉴定，随后用噻唑蓝（MTr）比色法绘制体外生长曲线并计算体外细胞群体倍增时间，用噻唑蓝（MTT）法检测30、60、90、120min黏附率，用Transwell法检测16h迁移率和侵袭率，用流式细胞术检测细胞周期和增殖指数及凋亡指数。结果CT26CEA细胞株与CT26GFP细胞株比较，30、60、90、120rain黏附促进率分别为10．0％（P〉0．05）、13．8％（P〈0．01），14．7％（P〈0．01），4．0％（P〉0．05），16h迁移促进率为21．8％（P〈0．01），侵袭促进率为39．5％（P〈0．01），体外细胞群体倍增时间、细胞周期分布、增殖指数、凋亡指数差异无统计学意义（P〉0．05）。结论癌胚抗原过表达可以促进结肠癌细胞黏附、迁移和侵袭，对细胞增殖和凋亡无显著影响。

Objective To investigate the effect of carcinoembryonic antigen （CEA） overexpression on the biological characteristics of colon cancer ceils. Methods The murine colon cancer CT26 ceils were transfected by recombinant lentivirus conjugated respectively with CEACAM5-eGFPcDNA and eGF- PcDNA to establish CT26CEA cell strain stably expressing CEA or control CT26GFP cell strain only ex- pressing enhanced green fluoresce protein. The expression of CEACAM5 mRNA and protein was detected by using reverse transcription polymerase chain reaction （RT-PCR）, and Western blotting, respectively. Methyl thiazolyl tetrazolium （MTF） assay was used to draw cell growth curve, and examine the cell adhe- sion at 30, 60, 90 and 120 rain and migration proliferation at 16 h. Flow cytometry was used to determine apoptosis rate , and Transwell chambers were used to measure the invasion ability of ceils. Results The CT26CEA cell strain showed positive adhesion promotion rate by 10. 0% （P〉 0. 05）, 13.8% （P 〈 O. 01 ）, 14. 7% （P 〈0. 01 ）, and 4. 0% （P 〉0. 05） respectively at 30, 60, 90 and 120 rain, positive mi- gration promotion rate by 21.8% （P 〈0. 01 ） and positive invasion promotion rate by 39. 5% （P 〈0. 01 ） respectively at 16 h, but no significant difference was found in the cell cycle between CT26CEA cell strain and CT26GFP cell strain （P 〉 0. 05）. Conclusion CEA can promote adhesion, migration, and invasion, while has no direct impact on proliferation and apoptosis of colon cancer cells in vivo.