重组双歧杆菌HSV-1TK/GCV联合抑制肝癌的研究

Research on the Effects of Anti-liver Cancer by Bifidobacterial Recombinant Herpes Simplex Virus Thymidine Kinase Combined with Ganciclovir

双歧杆菌被用来作为实体瘤靶向治疗的载体得到广泛应用。本研究探索重组双歧杆菌TK联合GCV(BFTK/GCV)治疗肝癌小鼠模型的效果。构建重组质粒p ET32-TK、p ET32-TK83,表达的6×His-TK、6×His-TK83融合蛋白与d T、d U、GCV作用,剩余产物用HPLC法检测。裸鼠腋下注射肝癌细胞株Hep G2细胞,构建裸鼠腋下肿瘤模型。药物处理后,测量肿瘤大小,绘制肿瘤生长曲线。已成功构建重组质粒p ET32-TK、p ET32-TK83。成功表达6×His-TK、6×His-TK83融合蛋白。TK83融合蛋白组的d U峰面积较大。成功构建裸鼠腋下肝癌肿瘤模型,BFTK/GCV明显抑制肿瘤的生长,治疗组和对照组肿瘤体积的差异有统计学意义。

Bifidobacterial was widely used as the vector of targeted therapy of solid tumors. The study in order to evaluate the effect of bifidobacterial recombinant HSV-1 TK combine with GCV(BFTK/GCV) on hepatic cancer.The TK gene and TK83 gene were cloned into the plasmid p ET32. The 6×His-TK fusion protein and 6×His-TK83 fusion protein were purified with Ni-NTA resin and used to be reacted with d T, d U and GCV, the remaining product was detected by HPLC. Naked mouse hepatic cancer model was established with hepatic cancer cell Hep G2 by injected into the oxter of nude mice. The tumor growth curve were drawn after 2w treatment by BFTK/GCV. Recombinant plasmid p ET32-TK and p ET32-TK83 were constructed successfully. The expressed fusion protein 6×His-TK and 6×His-TK83, with a relative molecular mass of about 60 k D. The peak area of d U in group 6 ×His-TK83 fusion protein was higher than that in 6 ×His-TK fusion protein group. Naked mouse hepatic cancer model was established successfully. BFTK/GCV significantly inhibited tumor growth, the difference of tumor volume between the treatment group and the control group was statistically significant.