Smad4对人胰腺癌细胞HS766T生长抑制的研究

Inhibitive effect of Smad4 gene on the growth of human pancreatic adenocarcinoma cell line HS766T

目的 探讨Smad4基因在人胰腺癌细胞中的肿瘤抑制作用。方法 应用分子克隆技术构建Smad4真核表达载体 ,然后用脂质体法将其导入到Smad4同源缺失的人胰腺癌细胞HS766T中 ,经G418筛选获得可稳定表达Smad4的人胰腺癌细胞克隆 ,用细胞计数和噻唑蓝 (MTT)法观察细胞生长速度及增值率 ,流式细胞仪 (FCM)检测Smad4表达、细胞凋亡及细胞周期情况。结果 获得了Smad4真核表达质粒pcDNA3 .1 Smad4。Smad4导入人胰腺癌HS766T细胞后 ,肿瘤细胞增值能力明显受到抑制、生长速率显著下降 ,并出现细胞周期G1期阻滞。结论 Smad4基因具有抑制胰腺癌细胞增殖的作用 。

Objective To study the inhibitory effect of Smad4 gene on the growth of human pancreatic adenocarcinoma cell line(HS766T)cells. Methods Smad4 cDNA was cloned into the pcDNA3.1 vector to construct a recombinant expression vector plasmid pcDNA3.1 Smad4, which was then transfected by liposome method into HS766T cells with homozygous deletion of Smad4. After G418 selection, Smad4 gene expression, cell cycle and cell apoptosis were assessed by flow cytometry, and the cell growth rate by cell count and MTT analysis. Results The Smad4 gene recombinant expression vector was constructed. After transfection with pcDNA3.1 Smad4, the proliferation of cultured HS766T pancreatic adenocarcinoma cells was suppressed with the suppression rate higher than 40% on the fourth day. The same results were found by MTT assay. The cells of G 1 phase increased obviously while the cell number of S phase decreased, although no obvious increase of apoptosic cell number was observed. Wild type Smad4 gene attributed to the increase of G 1 phase cells and the decrease of S phase cells in HS766T cells and could inhibit the growth of HS766T cells. Conclusion Wild type Smad4 gene can inhibit the proliferation of human pancreatic adenocarcinoma cells and can be used as one of the target genes of pancreatic adenocarcinoma gene therapy.