HSV-TK/GCV联合IL-2对人舌癌移植瘤的抑制作用

Inhibitive effect of HSV-TK and IL-2 on transplanted tumor of human tongue cancer

目的观察HSV-TK联合IL-2对人舌癌细胞生长的抑制作用,并初步探讨其作用机理。方法建立舌癌Tca8113移植瘤动物模型,采用HSV-TK基因联合IL-2基因对人舌癌移植瘤进行裸鼠体内实验。实验分A、B、C、D四个实验组,每组5只裸鼠。A组为对照组,每瘤注射生理盐水;B组为空病毒组,每瘤注射空病毒5×108PFU;C组为单纯TK治疗组,每瘤注射TK5×108PFU;D组为TK联合IL-2治疗组,每瘤注射TK(5×108PFU)+IL-2(5×108PFU)。48小时以后,对B、C、D组裸鼠每日腹腔注射GCV,100mg/kg/日,2次/日,连续10天。最后一次给药后第二天处死裸鼠,测量肿瘤重量并计算抑瘤率;光镜观察移植瘤组织学变化,透射电镜观察细胞的超微结构。结果A、B、C、D组裸鼠瘤重量均数分别为:1.52±0.49g,1.44±0.42g,0.91±0.24g,0.36±0.24g。C组的抑瘤率为40%;D组抑瘤率为76%。经统计学检验发现TK组和TK联合IL-2组的肿瘤生长均受到抑制,A组和C组、A组和D组之间有显著性差异(P<0.01)。与单用TK组比较,TK联合IL-2组抗瘤作用明显增强。光镜观察发现,TK组肿瘤组织呈灶状凋亡,可见核固缩的细胞。TK联合IL-2组凋亡细胞增加,肿瘤细胞减少。电镜观察发现,TK组和TK联合IL-2组出现大量的细胞凋亡:凋亡细胞皱缩,胞核体积缩小,染色质浓缩,电子密度增加,呈新月状边集于核膜下。结论TK联合IL-2治疗可显著抑制Tca8113移植瘤的生长,其治疗效果优于TK单独应用。

Objective To observe the inhibitive effects on tongue cancer by union treated with HSV -TK and IL -2 and to discuss its mechanism. Methods Animal models of nude mice loaded on Tca8113 tongue cancer were set up. Transference of HSV - TK and IL - 2 gene in vivo was done on transplanted tumors. The experiments involved 20 nude mice, divided into four groups A, B, C, D of 5 mice each. Group A was used as the control, and injected by normal saline; group B was used as the virus vector control, and injected by the virus vector 5 × 10^8PFU; group C was the / experimental group, and injected by TK 5 × 10^8PFU; group D was another experimental group, and injected by TK （5× 10^8pF） ＋ IL-2 （5× 10^8pFU）. After 48h, GCV abdominal injection was given to nude mice of group B, C, D, 100mg/ kg/daily, twice every day, for 10 days. The following day after the last medlelne was given, all nude mlee were saerliieed. The transplanted tumors were measured and observed by light microscope and transmission electron microscope （TEM）. Results The weights of the transplanted tumor of group A, B, C, D were 1.52 ±0.49g,1.44 ±0.42g,0.91 ±0.24g, 0.36 ± 0.24g, respectively. The ratio of tumor inhibition was 40% of group C and 76% of group D. The weighs of transplanted tumor were smaller in group C and group D than that in group A and group B （ P 〈 0.01 ）. There were significantly difference between any two groups （ P 〈 0.01 ）, exception group A and group B. The restrlts of light microscope in TK group revealed small apoptotie region and shrunk cells. There were not only many apoptotie cells in TK ＋ IL-2 group, but also extinct tumor cells. The restults of TEM in TK and TK ＋ IL-2 group revealed many apoptotie cells with their cytoplasm shrinkage,the chromatin condensed into block , and dispersed unevenly. A volume of apoptotie bodies were formed. Conclusion HSV- TK/GCV and IL- 2 eonld suppress the growth of Tea8113 transplanted tumors. The effect of HSV -TK ＋ IL- 2 on Tea8113 transplanted tumors was much better than that of HSV- TK alone.