益气解毒方干预鼻咽癌细胞裸鼠移植瘤肿瘤微环境细胞免疫耐受现象的初步研究

Reliminary study on the effects of Qi-Boosting Toxin-Resolving Formula on the phenomenon of immune tolerance in the tumor microenvironment of grafted tumor with nasopharyngeal carcinoma cells among nude mice

目的通过观察益气解毒方对鼻咽癌细胞裸鼠移植瘤的干预效应,探讨调节性T细胞诱导的鼻咽癌肿瘤微环境免疫耐受性特征及其对干预药物的反应性。方法人鼻咽癌细胞株CNE2细胞建立裸鼠移植瘤模型,然后随机分为模型组、益气解毒方治疗组、顺铂阳性对照组、益气解毒方与顺铂联合治疗组,每组8只动物,分别给予相应处理药物,系统观察各组裸鼠移植瘤生长情况及体积变化。观察期满后处死动物,取瘤体称重,分别计算抑瘤率,并切取瘤体组织标本,观察比较各组移植瘤的病理组织学变化,免疫组化法检测各组标本CD4、CD8、Foxp3表达活性,比较其组间差异。结果各组检测结果比较显示,益气解毒方处理后,移植瘤组织中的CD4、FoxP3表达活性受到明显抑制,CD8表达则明显提高,尤以联合治疗组表现明显,与模型组比较,组间差异具有显著性统计学意义。结论益气解毒方能有效改善鼻咽癌裸鼠移植瘤肿瘤微环境的免疫耐受现象而抑制肿瘤生长速度;联合应用化学药物后,该一效应尤为明显。

Objective To investigate the effect of Qi Boosting Toxin Resolving Formula （QBTRF） on the phenomenon of immune tolerance in the microenvironment of grafted tumor with nasopharyngeal carcinoma （NPC） cells （HNE2） among nude mice based on an in vivo experiment. Methods 2Thirty two BALB/c nude mice were used in this study to prepare grafted tumor models with human NPC cell line CNE2 by subcutaneously injecting method at first. Then, modeling mice were randomly divided into four groups, i.e. modeling group （MG） treated by orally taken normal saline, treating group （TG） treated with concentrated QBTRF solution, integrative treating group （ITG） with concentrated QBTRF solution and DDP injected peri／tonearly, and positive control group （PCG） treated by peri／tonearly injected DDP only, with 8 animals in each group. They were treated with corresponding treating procedures for 14 days respectively, when grafted tumors growing to a volumne about 1 cm in their diameter. Followed was observation on the dynamically growing status of grafted tumors during the experimental period among all groups. By the end of experimental period, all mice were put into death to dissect tumors for measuring the weight of grafted tumors to calculate the rate of tumor growinainhibiting and to take tissue samples from the dissected tumors for histopathological observation under light microscope respectively. Meanwhile, immunohistochemistry was made to detect the expressive levels of CD4,CD8 and Foxp3 in the cells of tumor tissues from all groups of animals. These results were also further confirmed by the procedures of Western blots. Results The inhibitory rates of tumor growing were 18.70%, 58.40% and 43.80% for TG, ITG and PCG respectively.Asdetermined by SABC immunohistochemistry followed by image analysis on these groups, the expressive activities of CD4 and Foxp3 were inhibited significantly in the samples of TG, ITG and PCG, while that of CD8 were obviously elevated in the tisuue samples of them （P0.01）. These results were confirmed by the data of Western blotting assays Conclusion Qi Boosting Toxin Resolving Formula, especially when integreted with DDP, can effectively reverse the phenomenon of immune tolerance in the grafted tumor microenvironment of NPC among nude mice to induce a highly elevated tumor-inhibiting rate, with significantly improved quality of life.