论文部分内容阅读
目的探讨NK细胞输入后在荷Heps肝癌小鼠体内的分布规律,光动力疗法(PDT)对输入的NK细胞在肿瘤组织内分布的影响。方法 96只KM小鼠建立Heps肝癌移植瘤模型后随机分为对照组、细胞组(NK组)和光免疫组(PIT组),每组32只。对照组小鼠尾静脉输入生理盐水,NK组小鼠尾静脉输入4,’6-二脒基-2-苯基吲哚(DAPI)标记的NK细胞,PIT组小鼠PDT处理后即刻输入DAPI标记的NK细胞。治疗后1、2、4、6和8 d摘取小鼠肝脏、脾脏、肺脏和肿瘤组织,在荧光显微镜下观察NK细胞在上述脏器的动态分布情况;治疗后2 d观察瘤组织的病理学变化;治疗后1、2、4、6和8 d摘除小鼠眼球取血以流式细胞仪检测外周血NK细胞百分率;治疗后6 d LDH释放法检测小鼠脾细胞杀伤活性。结果 (1)各治疗组,输入后1、2、4、6和8 d,NK细胞在小鼠肝脏、脾脏、肺脏和肿瘤组织中均有分布,其中以肿瘤组织内最多;NK细胞在瘤内浸润密度:以输入后2 d最高,而各时间点PIT组又显著高于NK组(P<0.05);(2)外周血中NK细胞含量:在治疗后1、2、4和6 d,PIT组较对照组明显增高(P<0.01);治疗后1、2和4 d,NK组较PIT组和对照组增高(与PIT组相比,P<0.05;与对照组相比,P<0.01);(3)效靶比为10∶1、20∶1和50∶1时,NK组和PIT组脾细胞杀伤活性均显著高于对照组,其中PIT组又显著高于NK组(P<0.01)。结论 NK细胞输入荷瘤小鼠体内后能选择性聚集于肿瘤组织内;PDT可增加输入的NK细胞在瘤内的浸润密度,增强小鼠脾细胞对Heps肝癌的杀伤活性。
Objective To investigate the distribution of NK cells in Hep Hepatic Carcinoma-bearing mice and the effect of photodynamic therapy (PDT) on the distribution of NK cells in tumor tissues. Methods Ninety-six KM mice were randomly divided into control group, NK group and PIT group with 32 mice in each group. The rats in the control group were given physiological saline through the tail vein, NK cells were labeled with 4,6-diamidino-2-phenylindole (DAPI) in the tail vein of the mice in the NK group, and DAPI Labeled NK cells. The livers, spleens, lungs and tumor tissues of mice were harvested at 1, 2, 4, 6 and 8 days after treatment, and the dynamic distribution of NK cells in the organs was observed under a fluorescence microscope. The tumor tissues were observed 2 days after treatment Neonatal mice were sacrificed at 1, 2, 4, 6, and 8 days after treatment. The percentage of NK cells in peripheral blood was detected by flow cytometry. The LDH release assay was used to detect the cytotoxic activity of mouse spleen cells 6 days after treatment. Results (1) NK cells were distributed in the liver, spleen, lung and tumor tissues at 1, 2, 4, 6 and 8 d after treatment in each treatment group, The infiltration density in infiltration was the highest at 2 d after infusion, and PIT at each time point was significantly higher than that in NK group (P <0.05). (2) The content of NK cells in peripheral blood at 1, 2, 4 and 6 d , PIT group was significantly higher than the control group (P <0.01); 1, 2 and 4 days after treatment, NK group increased compared with PIT group and control group (PIT group compared to P <0.05; compared with the control group, P <0.01). (3) When the target ratios were 10: 1, 20:1 and 50:1, the cytotoxic activity of spleen cells in NK group and PIT group were significantly higher than that in control group, PIT group was significantly higher than that in NK group P <0.01). Conclusion NK cells can be selectively aggregated into tumor tissues after they are introduced into tumor-bearing mice. PDT can increase the infiltration density of NK cells in the tumor and enhance the killing activity of mouse spleen cells on Heps liver cancer.