三元复合因子Net对人胰腺癌细胞株BxPC3增殖的影响

目的研究三元复合因子Net转染人胰腺癌细胞株BxPC后的表达状态及对原癌基因c—fos表达的影响。方法采用脂质体2000将重组人pEGFP—Net质粒和空质粒pEGFP转染人胰腺癌BxPC3细胞株，建立稳定高表达Net的细胞系。应用MTT、流式细胞仪等方法检测胰腺癌细胞的增殖，应用实时定量PCR和Western blotting检测Net及c—fos mRNA和蛋白的表达。结果BxPC3细胞低表达Net，转染pEGFP-Net后稳定高表达Net，并抑制c-fos的表达。转染pEGFP—Net的细胞生长缓慢，转染后第3、5、7天的抑制率分别为38．8％、55．3％、56．9％，显著高于空质粒转染组的5．1％、12．4％、8．6％（P〈0．05）；G0／G1期细胞占（61．8±5．7）％，显著高于空质粒转染组的（45．1±3．4）％。结论三元复合因子Net能抑制人胰腺癌BxPC3细胞的增殖，其机制可能与抑制c—fos表达有关。

The effect of ternary complex factor Net on the proliferation of human pancreatic carcinoma cell line BxPC3

Objective To investigate the expression of the ternary complex factor Net in human pancreatic carcinoma cell line BxPC3 and its effect on cell proliferation and the expression of c-fos. Methods pEGFP-Net prokaryotic expression plasmid and empty vector pEGFP were transfed into BxPC3 cells by using lipofectamine 2000, then monoclonal cell which stably expressing Net was established. Human pancreatic carcinoma cells proliferation was detected by MTT and flow cytometry. The mRNA and protein expression of Net and c-fos in BxPC3 cells were detected by real-time PCR and Western blot. Results Net was low expressed in BxPC3 cells. After pEGFP-Net transfeetion, Net was stably expressed and the expression of c-los was inhibited, cell proliferation was also inhibited after pEGFP-Net transfeetion, the inhibitory rates at the 3rd, 5th, 7th day was 38.81% , 55.34% and 56.92% respectively,which were significantly higher than those in the empty vector group （5.09% , 12.42% , 8.6% , P 〈 0.05 ）. G0/G1 phase cell was （ 61.79 ± 5.67） %, which were significantly higher than （45.14 ±3.37 ） % in the empty vector group （ P 〈 0.05 ）. Conclusions The ternary complex factor Net could inhibit pancreatic carcinoma cell line BxPC3 proliferation. Its mechanism was possibly repressing expression of oncogene c-fos.