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目的纯化人白细胞介素24的原核重组表达质粒pET-21a(+)-IL-24在大肠杆菌BL(DE3)中的表达产物rhIL-24,并观察重组表达蛋白rhIL-24在体外对胃癌细胞株SGC-7901凋亡的影响。方法IPTG诱导原核表达载体pET-21a(+)-IL-24在大肠杆菌中表达rhIL-24,经纯化、复性后作用于人胃癌细胞株SGC-7901。MTT法检测rhIL-24对细胞生长的影响,流式细胞仪检测凋亡峰,荧光染色观察细胞凋亡形态。结果rhIL-24对体外培养的胃癌细胞SGC-7901有明显的生长抑制作用,且呈时间、剂量依赖性。rhIL-24蛋白30μg/ml作用于SGC-7901细胞24h后,流式细胞仪检测结果显示有明显的凋亡峰出现,荧光染色后显示染色质固缩,细胞核呈致密浓染或呈碎块状致密浓染。结论本实验成功纯化IL-24原核表达产物rhIL-24,对体外培养的胃癌细胞株SGC-7901有明显的生长抑制、诱导凋亡作用。
Objective To purify rhIL-24 from the prokaryotic recombinant plasmid pET-21a (+) - IL-24 of human interleukin-24 in E.coli BL (DE3) and observe the effect of rhIL-24 recombinant protein on gastric cancer cells Effect of strain SGC-7901 on apoptosis. Methods IPTG induced prokaryotic expression vector pET-21a (+) - IL-24 to express rhIL-24 in Escherichia coli. The recombinant plasmid was purified and refolded and then applied to human gastric cancer cell line SGC-7901. MTT assay rhIL-24 on cell growth, apoptosis detected by flow cytometry, fluorescence staining observed apoptotic morphology. Results rhIL-24 had obvious growth inhibitory effect on SGC-7901 cells cultured in vitro in a time-and dose-dependent manner. After treated with rhIL-24 (30μg / ml) for 24 hours, the apoptotic peak appeared after flow cytometry analysis. The chromatin showed chromatin condensation after staining, and the nucleus was densely stained or broken Dense thick dye. Conclusions The recombinant prokaryotic expression product rhIL-24 of IL-24 was successfully purified in this experiment, which has obvious growth inhibition and induction of apoptosis in gastric cancer cell line SGC-7901 cultured in vitro.