ATRA对胃癌细胞株MKN-45的诱导分化及其相关酶活性的影响

Effect of ATRA on Differentiation and Related-Enzyme Activities of Gastric Carcinoma Cell Line MKN-45

目的 研究维甲酸核内受体 β(RARβ)基因转染后 ,全反式维甲酸 (ATRA)对低维甲酸反应性胃癌细胞系MKN 4 5的诱导分化及其相关酶活性的影响。方法 脂质体介导真核表达载体pCMV script RARβ转染MKN 4 5细胞系 ,筛选得到阳性克隆 ,Westernblot鉴定后给予RARβ受体相应配体ATRA ,四唑盐比色实验、5 溴脱氧尿嘧啶法测定细胞增殖水平 ,流式细胞仪测定细胞周期分布情况 ,酶标仪测定相关分化标志酶的活性。结果 ATRA可使高表达RARβ受体的MKN 4 5细胞系增殖速度减慢 ,细胞周期出现G0 /G1停滞 ,分化标志酶LDH、ALP和 β G的活性下降。结论 RARβ受体基因转染并结合使用相应配体ATRA可使MKN 4 5胃癌细胞系增殖减慢 。

Purpose. To study the effect of ATRA on differentiation and relatedenzyme activity of low-RA-responsive gastric carcinoma cell line MKN-45 transfected with RAPβ gene. Methods. Eukaryotic expression vector pCMV-script-RARβ was transfected into MKN-45 cell line mediated by Lipofectin AMINE, positive cell clone was screened and verified by Western blot. The proliferation of transfected MKN-45 was assayed by MTT as well as BrdU methods. Their cell cycles were detected by flow cytometer, the related-enzyme activities were analyzed by enzyme reader. Results. ATRA could inhibit the proliferation of MKN-45 cell line with high expression of RARβ cause cell cycle to arrest in G0/G1 phrase and decrease the activities of relative enzymes indicating cell differentiation. Conclusions. Transfecion of RARβ gene as well as using corresponding ligand ATRA could inhibit proliferation and deteriorate phenotype in gastric carcinoma cell line MKN-45.