粘蛋白1介导的抗胃癌细胞MKN-45侵袭作用的研究  

The role of mucin gene 1 mediated anti-MKN-45 cell invasion

作　　者：陶春华[1] 陈腾飞[1] Praveen K.Yadav 邬瑞金[1] 邱骅婧[1] 吴维[1] 刘占举[1] 

机构地区：[1]上海同济大学附属第十人民医院胃肠内科,200072

出　　处：《中华消化杂志》2012年第3期175-179,共5页Chinese Journal of Digestion

摘　　要：目的构建粘蛋白1（MUCl）基因的反义肽核酸（PNA），观察对胃癌细胞MKN一45侵袭能力的影响，并探讨其机制。方法根据MUC1基因的序列设计反义PNA序列，经脂质体介导转染胃癌细胞MKN45，并设空载体组（随机对照）和空白对照组（阴性对照）运用荧光定量PCR检测MUC1的表达，并观察E-粘附素的表达变化；TransWell小室实验观察对胃癌细胞侵袭力的影响。结果构建的3个MUCl基因的反义PNA均能有效抑制该基因表达，其表达水平分别为0．62±0．18，0．49±0．12和0．60土0．21，显著低于阴性对照组（1．18±0．03，P〈0．01）。阴性对照组与随机对照组之间差异无统计学意义（1．00±0．04，P=0．657）。取抑制效率最高的PNA进行后续研究转染MUClPNA后的胃癌细胞侵袭能力显著下降（t=2．09，P=0．005），并伴有E-粘附素基因及蛋白的表达上调。结论胃癌细胞MKN-45中MUCl基因与E_粘附素的表达存在负相关，抑制MUCI基因的表达能显著抑制肿瘤细胞的侵袭能力。Objective To create Mucin gene 1 （MUC1） antisense peptide nucleic acid （PNA）, and to observe its effects on MKN-45 cell invasion and explore the mechanism. Methods The sequence of antisense PNA was designed according to MUC1 gene sequence and transfected into human gastric cancer cells （MKN-45） by liposome, and the empty vector group （randomized control group） and blank control group （negative control group） were involved. The expression of MUC1 was detected by real time quantitative PCR and the changes of E^cadherin expression were also observed. The effects on gastric cancer cell invasion were tested with transwell chamber assays. Results The expression of MUC1 gene was effectively suppressed by the 3 created antisense PNA, and their expression level （0.62 ＋ 0.18,0. 49 ztz 0. 12 and 0. 60 -I- 0. 21 ） was significantly lower than that of negative control group （1. 18 4-0. 03, P ~ 0. 01）. There was no significant difference between radomized control group and negative control group （1.00±0.04, P=0. 657）. After MUC1 PNA transfected, the capability of gastric cancer cell invasion decreased significantly （P= 0. 005）. And the expression of E-cadherin at mRNA and protein level was up-regulated. Conclusions There is negative correlation between MUC1 and E-eadherin expression in gastric cancer cell MKN-45. The capability of tumor cell invasion is significantly inhibited by suppressing MUC1 gene expression.

关 键 词：胃肿瘤 肽核酸类 寡核苷酸类 反义 粘蛋白类 肿瘤细胞 培养 肿瘤浸润 

分 类 号：R735.2[医药卫生—肿瘤]