15-羟基前列腺素脱氢酶基因转染抑制鼠胃癌细胞MFC增殖的观察  

Inhibitory effect of 15-hydroxyprostaglandin dehydrogenase gene on the growth of gastric cancer cell MFC

作　　者：李丽华[1,2] 汪晓洁[1,2] 胡静[1,2] 杨立波[1,2] 寿涛[1,2] 

机构地区：[1]云南省第一人民医院肿瘤内科 [2]昆明理工大学附属医院肿瘤内科,云南昆明650032

出　　处：《中华肿瘤防治杂志》2012年第23期1785-1789,共5页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的:构建pcDNA3.1/15-PGDH(15-羟基前列腺素脱氢酶)真核表达载体,并转染鼠胃癌细胞MFC,观察15-PGDH对MFC细胞增殖的影响。方法:通过基因重组技术构建真核表达载体pcDNA3.1/15-PGDH,转染胃癌细胞MFC,建立稳定转染细胞株,RT-PCR检测转染细胞中15-PGDH基因的表达情况;MTT法测A值,绘制细胞生长曲线;克隆形成实验分析15-PGDH对胃癌细胞增殖能力的影响。结果:15-PGDH基因转染成功,转染后鼠胃癌MFC/15-PG-DH细胞中15-PGDH基因的相对表达量为1.06±0.08,明显高于空质粒转染组的0.22±0.01及未转染细胞组的0.21±0.01,差异有统计学意义,P<0.01。且基因转染后细胞生长明显受到抑制,MTT结果显示,细胞生长至第4、6、8天,15-PGDH基因转染组细胞的A值均较低,分别为1.173±0.072、1.405±0.040和1.689±0.116,与空质粒转染组细胞的1.452±0.062、1.955±0.098和2.755±0.324相比,差异有统计学意义;与未转染MFC细胞的1.534±0.082、1.972±0.073和2.477±0.319相比,差异亦有统计学意义,P值均<0.05。15-PGDH基因转染细胞组克隆形成率为18%,与未转染细胞组63%及空质粒转染细胞组59%相比,克隆形成明显受到抑制,差异有统计学意义,P<0.01。结论:15-PGDH基因转染可显著抑制鼠胃癌MFC细胞的增殖和克隆形成能力。OBJECTIVE：To construct eukaryotic expression vector pcDNA3.1/15-PGDH,transfect rat gastric cancer cell MFC,and observe the effect of 15-hydroxyprostaglandin dehydrogenase gene on the growth of MFC cell. METHODS： The eukaryotic expression vector peDNA3.1/15-PGDH was constructed by gene recombination. The recombinant plasmid was transfected into the gastric cancer cell line MFC,and a stably transfected cell line was established. The expression of 15-PGDH gene in MFC/15-PGDH cells was detected by RT-PCR assay; The cell growth curve was made by the result of MTT test The effect of 15-PGDH on the cancer cells was analized by the result of colony formation. RESULTS： The transfection of 15-PGDH gene was successful. The expression of 15-PGDH gene in MFC/15 PGDH cells was 1.06±0.08 after transfection, which was higher than that in MFC/pcDNA3.1 cells （0.22± 0.01）, and MFC ceils （0.21 ± 0.01）, and there was significant difference （P〈0.01）. MTT results showed that in the 6,8,10 days, the growth of MFC/15-PGDH cells was inhibited,the A value was 1. 173±0. 072,1. 405 ±0. 040 and 1. 689±0. 116. There was obvious differences, compared with MFC/pcDNA3.1 cells and MFC cells （P〈0.05）. The A value of MFC/pcDNA3. 1 cells was 1. 4524±0.062,1.955±0.098 and 2. 755±0.324. The A value of MFC cells was 1.534±0.082,1.972±0.073 and 2. 477±0. 319. The clone formation rate of MFC/15-PGDH cell was 18% ,it is decreased significantly compared with MFC/pcD NA3.1 cells and MFC cells,the clone formation rate of two laters were 63% and 59% and there was significant difference （P〈0.01）. CONCLUSION：The transfection of 15-PGDH gene significantly inhibite proliferation and colony formation of rat gastric cancer MFC cells.

关 键 词：胃肿瘤 15-PGDH 表达载体 转染 细胞增殖 基因表达 

分 类 号：R735.2[医药卫生—肿瘤]