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

The study on the lowered tumorigenicity of hepatocellular carcinoma cells cotransfected with chemokine mMIP-1α and costimulatory molecule m4-1BB L in vivo

目的观察巨噬细胞炎性蛋白-1α（MIP1α）联合4-1BB配体（4-1BBL）对肝癌细胞体内致瘤性的影响。方法以小鼠4-1BBL（m4—1BBL）重组逆转录病毒感染Hepa1-6小鼠MIP-1α（mMIP-1α），筛选并扩增抗性克隆，以流式细胞术检测m4—1BBL的表达，绘制并比较mMIP-1α和m4—1BBL同时或单独表达的Hepa1—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Hepa1-6mMIP-1α＋m41BBL、Hepa1-6m4—1BBL、Hepa1-6mMIP-1α、Hepa1-6pBabepuro、Hepa1-6、Hepa1-6pLXSHD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mMIP1α和m4—1BBL的小鼠肝癌细胞Hepa1—6mMIP—1α＋m4—1BBL，mMIP-1α和m4—1BBL同时或单独表达不影响Hepa16的生长曲线。观察5周，Hepa1-6mMIP-1α＋m4—1BBL接种的小鼠均未生长肿瘤，Hepa1-6mMIP-1α＋m4—1BBL的体内致瘤性低于Hepa1-6mMIP-1α和Hepa1—6m4—1BBL。接种Hepa1-6mMIP1α＋m41BBL的小鼠12周末存活率（9／9）高于接种Hepa1-6m41BBL小鼠（6／9）和Hepa1-6mMIP-1α小鼠（1／9）。结论趋化因子MIP-1α联合共刺激分子4-1BBL降低了肝癌细胞体内致瘤性，并使小鼠的生存期延长。

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-la and m4-1BB L were successfully established. The expression of mMIP-la 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 1-6 mMIP-1α＋m4-1BB L was lower than that of Hepa 1-6 mMIP-la 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-la injected mice （1/9）. Conclusion Chemokine MIP-la combined with eostimulatory 4-1BB L lowered the tumorigenicity of hepatoeellular carcinoma ceils in vivo, and prolonged the mice survival period.