食管癌引流区淋巴结细胞裸鼠体内抑瘤实验

In Vivo Tumor-inhibition in Esophageal Cancer Cells in Nude Mice

目的观察肿瘤引流区淋巴结(TDLN)细胞对自体瘤细胞体内杀伤作用。方法将TDLN细胞分为3组进行培养:TDLN-Ⅰ组培养基中添加IL-2,TDLN-Ⅱ组添加IL-2+IL-4+粒-单核细胞集落刺激因子(GM-CSF),TDLN-Ⅲ组添加IL-2+IL-4+GM-CSF+自身食管癌细胞抗原(tAg)。选择60只BALB/C裸鼠,用术中切取的食管癌组织块建立荷瘤裸鼠模型,将裸鼠随机分为空白对照组、IL-2组、TDLN-Ⅰ组、TDLN-Ⅱ组和TDLN-Ⅲ组,各6只,分别给予0.9%氯化钠注射液、IL-2及相应TDLN细胞。每周测量并记录一次裸鼠的肿瘤体积,5周后处死裸鼠取出移植瘤标本,用流式细胞术检测肿瘤细胞凋亡率。结果实验的不同时间5组裸鼠肿瘤体积间差异有统计学意义(F=30.020,P<0.01)。进一步分析显示:空白对照组与IL-2组肿瘤体积间差异无统计学意义(P>0.05);实验进行3、4、5周时TDLN-Ⅰ组、TDLN-Ⅱ组及TDLN-Ⅲ组肿瘤体积均较空白对照组及IL-2组显著降低,差异均有统计学意义(P<0.01);TDLN-Ⅰ组与TDLN-Ⅲ组间差异显著(P<0.05)。各组肿瘤细胞凋亡率间差异有统计学意义(P<0.01),TDLN-Ⅲ组的肿瘤细胞凋亡率最高为(39.1±1.5)%。结论TDLN细胞在体内具有杀伤肿瘤细胞和诱导肿瘤细胞凋亡的功能,可以作为一种新的治疗肿瘤的有效手段。

Objective To study in vivo tumor inhibition effects on tumor-draining lymph nodes （TDLN） cells in nude mice.Methods TDLN mice were cultured in 3 groups：IL-2 was added to the culture medium in TDLN-Ⅰ group,IL-2＋IL＋GM-CSF in TDLN-Ⅱ group,IL-2＋IL-4＋GM-CSF＋tAg in TDLN-Ⅲ group.The mice were divided randomly into groups blank control,IL-2,TDLN-Ⅰ,-Ⅱ,-Ⅲ,which were injected with NS,IL-2,TDLN cells,respectively.Tumor volume of nude mice were measured and recorded once per week.Mice were sacrificed to remove transplanted tumor samples and tumor cell apoptosis detected by flow cytometry.Results There was significant difference in tumor volume between 5 groups at different time of experiment （F=30.020,P〈0.01）.A further analysis showed that no significant difference was noted in tumor volume between groups control and IL-2 （P〈0.05）.Tumor volume decreased in groups TDLN-Ⅰ,Ⅱ,Ⅲ as compared with groups control and IL-2,the difference was significant （P〈0.01）,and TDLN-Ⅰ were significantly different from TDLN-Ⅱ （P〈0.05）.There was significant difference in tumor apoptosis rate between 5 groups （P〈0.01）,the apoptosis rate was the highest in TDLN-Ⅲ group （39.1±1.5）.Conclusion TDLN cells,having functions of aiti-tumor cells and inducing tumor cells to apoptosis,can be used as a new effective method to treat tumors.