摘要
目的观察应用反义脱氧寡核苷酸（ASODN）体外抑制MUC2对人胃癌细胞株SGC7901黏附活性的影响。方法应用人工合成的MUC2 ASODN经阳离子脂质体包裹后转染人胃癌细胞株SGC7901，采用逆转录-聚合酶链反应（RT-PCR）、免疫组织化学、玫瑰红法检测与内皮细胞黏附能力、噻唑蓝（MTT）法检测其与基质的黏附能力等方法体外观测对胃癌细胞黏附作用的影响。结果RT-PCR结果显示SGC7901细胞转染MUC2 ASODN 48h后，MUC2的mRNA（1．2380±0．1019）和蛋白（1．20±0．00）表达与对照组（1．8130±0．1651，6．50±0．45）比较均明显降低（P〈0．01、P〈0．05）；癌细胞与基质（0．7335±0．0679）及内皮细胞（0．0350±0．0224）的黏附能力与对照组（0．9072±0．1057，0．1262±0．0660）比较都有明显下降（P〈0．01）；CD44V6表达明显降低（1．50±0．09，对照组5．50±0．25），而E-钙黏蛋白表达增高（5．50±0．23，对照组3．50±0．28）（P〈0．05）。结论使用人工合成MUC2 ASODN能有效抑制胃癌细胞株SGC7901的黏附作用，降低肿瘤细胞的侵袭转移潜能。
Objective： To investigate the inhibitory effect in vitro of mucin gene MUC2 antisense oligodeoxynucleotide （ASODN） on its gene expression and cell adhesion ability on gastric cancer cells SGC7901. Methods Phosphorothioate MUC2 ASODN was synthesized and transfected to SGC7901 cells mediated by lipofectin,and its expression on SGC7901 cells was detected by RT-PCR. Its inhibitory effects on cell adhesion of SGC7901 cells to Matrigel after treated with MUC2 ASODN was measured by MTT method ,and adhesion to endothelial cells was measured by Rose Bengal. The expression of MUC2 protein, CD44V6 and E-cadherin protein on gastric cancer was detected by immunohistochemical method. Results As compared with the control cells （ 1.20 ± 0.00 ）, MUC2 ASODN could significantly inhibit the expression of MUC2 mRNA （ 1. 238 ±0.1019） of SGC7901 cells （ P 〈 0.01 ）. MTT method revealed that transfection of MUC2 ASODN into SGC7901 cells could significantly decreased the adherence ability to Matrigel of SGC7901 cells as compared with the control cells （P 〈 0.01 ）. Data of Rose bengal method suggested the number of the cells adherent to endothelial cells ECV304 was decreased （0. 1262±0. 0660） as compared with the control cells （0.0350 ±0. 0224, P 〈 0.01 ）. The immunohistochemistry indicated that transfection with MUC2 ASODN could down-regulate the expression of MUC2 protein （ 1.20 ± 0.0,6.50 ± 0.45 ） and CD44V6 （ 1.50 ±0.09, control 5.50 ±0. 25, P 〈 0. 05 ）, but up-regulate the expression of E- cadherin protein （5.50 ± 0.23, control 3.50 ± 0.28, P 〈 0.05 ）. Conclusion In vitro MUC2 ASODN treatment could significantly inhibit the MUC2 mRNA expression, cells adhesion ability to Matrigel and endothelial cells on gastric cancer cells SGC7901.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2008年第1期63-65,共3页
Chinese Journal of Experimental Surgery
基金
重庆市卫生局科研基金资助项目（渝卫科教2000-48）
关键词
胃癌
反义寡核苷酸
黏附
Gastric carcinoma
Antisense oligodeoxynucleotide
Adhesion
