人γδT细胞、CD3AK细胞和NK细胞对裸鼠移植人结肠癌的抑制作用及对肿瘤细胞凋亡的影响

The study of inhibition and apoptosis effects when using human γδT Cells, CD3AK Cells and NK Cells together in nude mice implanted with human colorectal cancer cells

目的 探讨人γδT细胞、CD3AK细胞和NK细胞对裸鼠移植人结肠癌的抑制作用及对肿瘤细胞凋亡的影响.方法 将人结肠癌细胞株（SW-116）接种于BALC /c裸鼠皮下,建立结肠癌裸鼠模型,随机分为4组,每组6只.设定建立模型为第1天,分别于第1天、第4天、第7天每组给予相应的治疗.A组：尾静脉注射RPMI 1640培养液;B组：注射NK细胞＋γδT细胞;C组：注射NK细胞＋CD3AK细胞;D组：注射NK细胞＋CD3AK细胞＋γδT细胞.并于第10天处死裸鼠,测量肿瘤体积,用Tunel法检测肿瘤细胞凋亡率.结果 24只裸鼠接种5×106结肠癌细胞后第4天,在接种部位均有肿瘤形成,皮下结节最大径为2～3 mm,成瘤率为100%.治疗组肿瘤体积增加小于对照组,第1次治疗后10天时测A、B、C、D组平均肿瘤体积分别为（86.78±7.19）、（52.80±5.59）、（54.05±6.53）、（43.16±7.40）mm3,各治疗组与对照组比较差异有统计学意义（P〈0.05）,B、C组组间无显著差异（P〉0.05）,D组与B、C组相比有显著差异（P〈0.05）.Tunel检测结果显示,与对照组相比,治疗组肿瘤细胞的凋亡率升高（P〈0.05）,以D组为甚.结论 人的γδT细胞、CD3AK细胞和NK细胞能不同程度延缓裸鼠移植的人结肠癌瘤体生长,免疫治疗能提高肿瘤细胞的凋亡率.

Objective To investigate the inhibition and apoptosis effects when using human γδT cells, CD3AK cells and NK cells together in nude mice implanted with human colorectal cancer cells. Methods We inoculated the colorec- tal cancer cell （SW - 1116） into 24 BALC/c nude mice. Then the mice were randmly divided into four groups. Group A was injected RPMI 1640 into vena caudalis as a control group, group B was injected NK cells ± γδT cells; group C was injected NK cells ± CD3AK cells; group D was injected CD3AK cells ± NK cells and ± γδT cells. We set the day when the module was established as the first day. The corresponding treatment was given on the first day, the fourth day, and the seventh day. Mice were killed on the tenth day. The transplanted tumor size of each mouse was computed and we also detected the apoptosis rate by TUNEL method. Results A tumor of 2 mm in diameter was formed in the inoculation site on the fourth day, the tumor formation rate was 100%. On the tenth day after treatment, the tumor average size of group A, B, C, and D were （86.78 ±7.19）, （52.80 ±5.59）, （54.05 ±6.53） , （43.16 ±7.40） mm^3, respectively. There was significant difference （P 〈 0.05 ） between control groups and treatment groups in tumor size. The difference was also exsited between D group and B, C groups （ P 〈 0.05 ） , but there was no significant difference between group B and C （ P 〉 0.05 ）. TUNEL assay also showed that the apoptosis rate was significantly increased in treatment gTOUpS compared to the control group （P 〈 0.05 ）, the increase trend was especially obvious in group D. Conclusion The human NK cells, CD3AK cells, γδT cells therapy can delay colorectal cancer growth in nude mice. The immunotherapy can inhibit cancer cell proliferation and promote apoptosis.