[婴儿双歧杆菌介导的CD和UPRT基因与5-氟胞嘧啶体外转染用于黑色素瘤的基因治疗]

[Gene therapy for melanoma by bifidobacterium infantis-mediated transfer of CD and UPRT genes with 5-FC in vitro].

作者信息

An Li-na, Li Zhu-hua, Yue Yang, Wang Shu-ren, Guo Zhi-ying

机构信息

Pathophysiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.

出版信息

Sichuan Da Xue Xue Bao Yi Xue Ban. 2007 Jan;38(1):27-30, 63.

PMID:17294721

Abstract

OBJECTIVE

This study was conducted by using Bifidobacterium Infantis as a delivery system to transport suicide gene CD and UPRT to tumors in an attempt to assess the UPRT-enhanced antitumor effect of CD/5-FC.

METHODS

The recombinant plasmid pGEX-UPRT was constructed and transfered into Bifidobacterium Infantis by electroporation and then identified. The synergistic antitumor effect of coexpression CD and UPRT was determined by MTT method. And the morphologic changes of B16-F10 cells were observed.

RESULTS

Recombinant Bifidobacterium Infantis could express UPRT correctly. The suvival rate of cells administrated CD+ UPRT and 5-FC was significantly lower than that of control (P<0.01), and the 5-FC sensitivity (IC50 = 0.015 micromol/mL) exhibited a 8. 5-fold increase when compared with that of cells administrated CD alone (IC50 = 0.127 micromol/mL). The cells treated with CD+UPRT were remarkably damaged morphologically, and the growth of cells was significantly inhibited as compared with that of other groups.

CONCLUSION

Recombinant Bifidobacterium Infantis with UPRT gene can significantly enhance the killing effect of CD/5-FC suicide gene system on melanoma B16-F10 cells of mice.

摘要

目的

本研究以婴儿双歧杆菌作为递送系统，将自杀基因CD和UPRT转运至肿瘤，旨在评估UPRT增强CD/5-FC的抗肿瘤作用。

方法

构建重组质粒pGEX-UPRT，通过电穿孔法将其转入婴儿双歧杆菌并进行鉴定。采用MTT法测定共表达CD和UPRT的协同抗肿瘤作用，并观察B16-F10细胞的形态学变化。

结果

重组婴儿双歧杆菌能够正确表达UPRT。给予CD+UPRT和5-FC处理的细胞存活率显著低于对照组（P<0.01），与单独给予CD处理的细胞相比，5-FC敏感性（IC50 = 0.015 μmol/mL）提高了8.5倍（IC50 = 0.127 μmol/mL）。经CD+UPRT处理的细胞在形态上受到显著损伤，与其他组相比，细胞生长受到明显抑制。