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NK细胞对人鼻咽癌细胞CNE2裸鼠皮下移植瘤的抑制作用

Effect of NK Cells on Tumor Growth Inhibition of Human Nasopharyngeal Carcinoma Cell (CNE2) Xenografts in Nude Mice

  • 摘要: 目的 研究同种异体NK细胞对人鼻咽癌细胞(CNE2)裸鼠皮下移植瘤的抑制作用。方法PCR-SSP法检测CNE2细胞HLA-A、B、Cw表型、NK细胞KIR表型(选择3例健康者为试验对象),磁珠分离法分离NK细胞并进行体外培养扩增,LDH释放法测定NK细胞对CNE2细胞的体外杀伤活性。12只BALB/c裸鼠分为两组,每组6只,对照组裸鼠每只皮下接种1×106CNE2细胞,治疗组裸鼠每只皮下接种1×106CNE2细胞,同时每只经尾静脉注入3×107NK细胞,观察两组裸鼠成瘤时间、成瘤率、肿瘤体积变化、计算抑瘤率。结果 CNE2细胞表面HLA-A、B、Cw表型为A2,24;B18,35;Cw4,7,3例健康者均表达KIR2DL1、KIR2DL3、KIR3DL1、KIR3DL2。效靶比5∶1、10∶1、20∶1、30∶1时,NK细胞对CNE2细胞的杀伤活性分别为(9.37±2.14)%、(27.14±1.82)%、(36.40±4.28)%and(54.67±2.80)%。对照组和NK细胞治疗组肿瘤出现时间分别为(10.00±2.68)d、(18.80±1.64)d,(P〈0.01),成瘤率分别为100%(6/6)、83.33%(5/6),对照组和NK细胞治疗组裸鼠的瘤重分别为(2.22±0.09)g、(1.42±0.09)g,(P〈0.01),NK治疗组的抑瘤率为36.04%。肿瘤组织石蜡切片病理学鉴定为低分化鳞状上皮细胞癌,NK细胞治疗组可见角化肿瘤细胞,较多的淋巴细胞浸润和大量细胞坏死区。结论 NK细胞对鼻咽癌裸鼠皮下移植瘤有明显的抑制作用,有希望成为治疗鼻咽癌的新方法。

     

    Abstract: Objective  To study the effect of natural killer (NK) cells on tumor growth inhibition of human nasopharyngeal carcinoma cell (CNE2 ) xenograft in BALB/c nude mice. Methods  HLA typing of CNE2 cells and KIR (killer cell immunoglobulin-like receptor) genotypes were determined by PCR-SSP. Cytotoxicity of NK cells (isolated f rom 3 healthy persons ) against CNE2 cells were detected by LDH releasing assay. 12 BALB/c nude mice were divided into two groups ( the control group and the treatment group), 6 BALB/c nude mice in the treatment group were injected subcutaneously 1 ×106 CNE2 cells together with 3 ×10 NK cells were injected intravenously into the tail veins, while the cont rol group were just injected 1 ×106 CNE2 cells subcutaneously. The changes of tumor volume were observed, and the tumor growth inhibition was calculated. Results  The HLA genotypes of CNE2 cells were A2, 24, B18, 35, Cw4, 7. KIR genotypes of the 3 healthy persons were KIR2DL1, KIR2DL3, KIR3DL1, KIR3DL2.Cytolysis of NK cells against CNE2 cells in vit ro were (9. 37 ±2. 14) %, (27. 14 ±1. 82) %, (36. 40 ±4. 28) % and (54. 67 ±2. 80) % respectively at 5∶1, 10∶1, 20∶1, 30∶1 E: T ratios. NK cells significantly inhibited the tumor growth in vivo, in the cont rol and treatment group, the rate of tumor formation were100 %(6/ 6 ), 83. 33 % ( 5/ 6 ), respectively ; the time of tumor formation were ( 18. 80 ±1. 64 ) d, (10. 00 ±2. 68) d, ( P < 0. 01) ; the average weight of tumors were (2. 22 ±0. 09) g, (1. 42 ±0. 09) g, ( P< 0. 01), respectively, The growth inhibitory rate was 36. 04 %. HE stain show that there were more necrosis regions and keratinization of tumor cells in the treatment group than that in the control group. Conclusion  Our findings suggest that NK cells can inhibit the human nasopharyngeal carcinoma xenografts in nude mice and may become a novel treatment approach for human nasopharyngeal carcinoma.

     

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