巨噬细胞炎性蛋白-1α联合4-1BB配体降低肝癌细胞体内致瘤性研究

目的 观察巨噬细胞炎性蛋白-1α(MIP-1α)联合4-1BB配体(4-1BB L)对肝癌细胞体内致瘤性的影响.方法 以小鼠4-1BB L(m4-1BB L)重组逆转录病毒感染Hepa 1-6小鼠MIP-1α(mMIP-1α),筛选并扩增抗性克隆,以流式细胞术检测m4-1BB L的表达,绘制并比较mMIP-1α和m4-1BB L同时或单独表达的Hepa 1-6细胞的生长曲线.C57B/L小鼠随机分为7组,每组9只,各组分别接种Hepa 1-6 mMIP-1α+m4-1BB L、Hepa 1-6 m4-1BB L、Hepa 1-6 mMIP-1α、Hepa 1-6 pBabe puro、Hepa 1-6、Hepa 1-6 pLXSHD和PBS,观察比较各组肝癌细胞的致瘤性,比较各组小鼠的存活率.结果 成功获得同时表达mMIP-1α和m4-1BB L的小鼠肝癌细胞Hepa 1-6 mMIP-1α+m4-1BB L,mMIP-1α和m4-1BB L同时或单独表达不影响Hepa 1-6的生长曲线.观察5周,Hepa 1-6 mMIP-1α+m4-1BB L接种的小鼠均未生长肿瘤,Hepa 1-6 mMIP-1α+m4-1BB L的体内致瘤性低于Hepa 1-6 mMIP-1α和Hepa 1-6 m4-1BB L.接种Hepa 1-6 mMIP-1α+m4-1BBL的小鼠12周末存活率(9/9)高于接种Hepa 1-6 m4-1 BB L小鼠(6/9)和Hepa 1-6 mMIP-1α小鼠(1/9).结论 趋化因子MIP-1α联合共刺激分子4-1BB L降低了肝癌细胞体内致瘤性,并使小鼠的生存期延长.

Abstract：

Objective To investigate the effects of macrophage inflammatory protein-1α (MIP-1α) combined with molecule 4-1BB L on the tumorigenicity of hepatocellular carcinoma cells in vivo. Methods Mouse MIP-1α (mMIP-1α) expressed Hepa 1-6 cells were transfected with m4-1BBL recombinant retrovirus, the anti-histidinol cells clones were selected and amplified. The expression of m4-1BB L was confirmed by flow cytometry. The growth curve of Hepa 1-6 cells transfected with mMIP-1α and m4-1BBL alone or together was drawn and compared. C57B/L Mice were randomly divided into 7 groups, 9 mice in each group, injected with mMIP-1α+m4-1BB L Hepa 1-6 cells, m4-1BB L Hepa 1-6 cells, mMIP-1α Hepa 1-6 cells, Hepa 1-6 cells, pLXSHD Hepa 1-6 cells or PBS respectively. The tumorigenicity of hepatocellular carcinoma cells and the mice survival rate were compared between each groups. Results Hepa 1-6 mMIP-1α+m4-1BB L cells which expressed both mMIP-1α and m4-1BB L were successfully established. The expression of mMIP-1α and m4-1BB L alone or together did not affect the growth curve of Hepa 1-6 cells. Observed for 5 weeks, no tumor developed in Hepa 1-6 mMIP-1α+m4-1BB L injected mice. The tumorigenicity of Hepa 1-6 mMIP-1α+m4-1BB L was lower than that of Hepa 1-6 mMIP-1α or Hepa 1-6 m4-1BB L in vivo. The survival rate of Hepa 1-6 mMIP-1α+m4-1BBL injected mice(9/9) was higher than that of Hepa 1-6 m4-1BB L injected mice (6/9)or Hepa 1-6 mMIP-1α injected mice (1/9). Conclusion Chemokine MIP-1α combined with costimulatory 4-1BB L lowered the tumorigenicity of hepatocellular carcinoma cells in vivo, and prolonged the mice survival period.