重组人白细胞介素24蛋白对胰腺癌细胞生长的影响

目的构建人白细胞介素24(IL-24)基因的表达载体并在大肠杆菌中诱导表达,观察纯化后的IL-24融合蛋白导致人胰腺癌细胞(Panc-1)生长抑制的作用。方法用限制性内切酶从质粒pET30b-IL-24中酶切获取人IL-24cDNA片段。并将该片段与pET42a原核表达载体基因重组,在大肠杆菌中实现重组基因的表达,提取并纯化rhIL-24蛋白。用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳法(SDS-PAGE)和Westernblot对表达产物进行鉴定。以不同浓度的重组蛋白IL-24与胰腺癌细胞Panc-1、正常肝胚细胞CCC-HEL-1共培养48h,同时设立相应浓度梯度的HIS-GST融合蛋白组对照组。采用四甲基偶氮唑蓝(MTT)法检测各组细胞的生长抑制情况。结果酶切结果证实,成功构建了pET42a-IL24原核表达载体,并在大肠杆菌中获得稳定的表达。rhIL-24蛋白以浓度依赖性的关系抑制Panc-1细胞的生长,抑制率明显高于HIS-GST蛋白。结论rhIL-24蛋白对胰腺癌细胞Panc-1生长有抑制作用。

Impacts of human interleukin-24 on the growth of pancreatic cancer cells

Objective To express human interleukin 24d （IL-24） gene in E coli using a constructed recombinant vector and to investigate the inhibitory effect of HIS-GST-IL-24 fusion protein on the growth of human pancreatic carcinoma cells （Panc-1）. Methods Human IL-24 cDNA fragments were amplified from plasmid pET30b-IL-24 by restricted endonuclease digestion and then cloned into the prokaryotic vector pET42a to express the recombinant gene in E coli. The expressed HIS-GST-IL-24 fusion protein was purified with GST-Sepharose 4B medium and identified by SDS-PAGE and Western blot. In vitro cultured human Panc-1 cells and normal CCC-HEL-1 cells were incubated with different concentrations of HIS-CST-IL-24 fusion protein for 48 hours, respectively, compared to the same treatments using corresponding concentrations of HIS-GIS fusion protein. The cell proliferation in each group was measured by MTr assay. Results Restricted endonuclease digestion showed successful construction of the recombinant prokaryotic expression vector pET42a-IL-24 and stable gene expression in E coli. rhIL-24 proteins inhibited the proliferation of Panc-1 cells in a concentration dependent manner, which was more potential as compared with HIS- GST proteins. Conclusion rhIL-24 protein appeared inhibitory against Panc-1 cell growth in a concentration-dependent manner.