摘要
目的探讨rhIL-24蛋白体外对胃癌生长的影响及作用机制。方法用已构建成功的真核表达载体pcDNA3.0-IL24质粒转染中国仓鼠卵巢细胞CHO细胞,使其上清稳定表达rhIL-24,同时用已构建成功的原核表达载体PET21a(+)IL-24-BL-21,抽提BL-21菌中表达的rhIL-24,行SDS-PAGE电泳和Western-Blotting鉴定rhIL-24的表达,MTT法检测rhIL-24对SGC7901胃癌细胞生长的影响;Hoechst染色、流式细胞术检测CHO细胞和BL-21菌表达的rhIL-24诱导胃癌细胞凋亡;鸡胚绒毛尿囊膜试验观察rhIL-24对血管形成的影响。结果rhIL-24对胃癌细胞的生长有明显的抑制作用;rhIL-24可诱导SGC7901胃癌细胞凋亡;rhIL-24具有抑制血管形成的作用。结论rhIL-24蛋白有多重抗肿瘤能力,其可能的作用机制是明显抑制胃癌细胞增殖,诱导肿瘤细胞凋亡,抑制血管生成。
Objective To approach the effect and its mechanism of anti-gastric cancer of the recombinant human IL-24（rhIL-24） in vitro. Methods The constructed eukaryotic expression vector of hIL-24 cDNA expressed in ClIO cells and the constructed prokaryotic expression vector of PET21a （ ＋ ）IL-24 expressed in BL-21 was made, then the CHO cells and the BL-21 to express rhIL-24 highly and stably was made. The expression of rhIL-24 in ClIO cells and in BL-21 were detected by SDSPAGE and western-blotting. The effects on the growth of SGC7901 gastric cancer line of recombinant protein hIL-24 were tested by MTT assay, and apoptosis-inducing activities by Hoechst ＆ FCM assay. The effect of rhIL-24 on angiogenesis was evaluated by chick embryo chorioallantoic membrane vascellum model. Results Expression of rhIL-24 in ClIO cells and BL-21 was identified by SDS- PAGE and western-blotting. The growth of SGC7901 was significantly inhibited by rhIL-24, the apoptosis of SGC7901 cells induced by rhIL-24 was proved by Hoechst ＆ FCM assay, and rhIL-24 could significantly inhibit the angiogenesis. Conclusion The protein of rhIL-24 revealed it＇ s multimechanisms of inhibiting gastric cancer growth. The mechanisms maybe include restraining the growth of SGC7901 cells, inducing the apoptosis of cancer cells and inhibiting the cancer-angiogenesis.
出处
《苏州大学学报（医学版）》
CAS
北大核心
2006年第3期370-374,共5页
Suzhou University Journal of Medical Science
基金
苏州大学医学发展基金(No.EE134517)
江苏省高校自然科学基金(05KJB320124)资助
