肝细胞癌患者外周血及肿瘤微环境中自然杀伤细胞的亚群变化及其临床意义

目的 通过检测不同自然杀伤细胞（NK细胞）亚群在肝细胞癌（HCC）外周血和肿瘤微环境中的变化规律，探讨NK细胞亚群与HCC疾病进展的关系。方法 流式细胞术检测40例HCC患者外周血、癌组织及癌旁组织中CD3-CD56+、CD3-CD57+、CD3-CD161+NK细胞亚群的含量，并收集36例健康志愿者的外周血作为对照。免疫组化法检测10例HCC癌组织和癌旁组织及3例肝良性肿瘤患者正常肝脏组织中CD3-CD56+NK细胞的分布。结果 流式细胞术检测显示，HCC组织CD3-CD56+和CD3-CD57+NK细胞比例分别（20.6±10.4）％ 和（6.9±3.7）％，低于癌旁组织中的（27.9±13.5）％和（9.6±5.1）％，差异有统计学意义（P＜0.05）；CD3-CD161+NK细胞在HCC癌组织和癌旁组织中含量的差异无统计学意义（P＞0.05）。3种NK细胞亚群在HCC患者外周血中的含量高于健康志愿者，但差异无统计学意义（P＞0.05）。免疫组化染色显示，癌旁组织中有大量CD3-CD56+NK细胞浸润，在HCC组织中只有少量分布，均少于在肝良性肿瘤的正常肝脏组织中的分布。Ⅰ+Ⅱ期HCC组织中NK细胞的比例高于Ⅲ+Ⅳ期，以CD3-CD56+及CD3-CD161+NK细胞为主。NK细胞不同亚群的比例与性别、甲胎蛋白、谷丙转氨酶、谷草转氨酶、TNM分期、白细胞计数、肝炎病毒感染、病灶数量无明显相关（P＞0.05）。结论 NK细胞比例下降与HCC患者的自然免疫下降及肿瘤进展有关。

The changes of NK cells in peripheral blood mononuclear cells and tumor microenvironment of hepatoceHu- lar carcinoma and their clinical significance

Objective To investigate the frequency of different subsets of natural killer （NK） cells in tumor microenvironment and peripheral blood mononuclear cells（PBMCs） from hepatocellular carcinoma（ HCC）, and discuss the relationship between NK cells subsets and HCC disease progression. Methods CD3-CD56＋, CD3-CD57~ and CD3-CD161~ NK cells were tested from fresh tissue samples and PBMCs by flow cytometry and peripheral blood of 36 healthy volunteers was collected as normal control. Ten HCC speci- mens were tested by immunohistochemical method to detect the corresponding distribution of CD3-CD56~ NK cells, and normal tissues from 3 benign hepatic tumors were collected as control. Results Flow cytometry detection showed that frequency of CD3-CD56~ cells and CD3-CD57~ cells were （20. 6＋ 10. 4）% and（ 6. 9＋3.7）% in 40 HCC tissues, lower than（ 27. 9-＋ 13.5）% and （9. 6-＋5.1 ）% in ad- jacent normal tissues（P〈O. 05）. And there was no significant difference in CD3-CD161~ cells（P〉0. 05）. The frequency of the 3 kinds of NK cells subsets in HCC PBMCs was higher than that in healthy volunteers, but without differences （P 〉 0.05 ）. Immunohistochemical staining showed that there was large amount of CD3-CD56~ NK cells infiltration in adjacent normal tissue of HCC and only a small distribution in tumor tissue, but they were less than CD3-CD56~ NK cells in normal tissues from 3 benign hepatic tumors. The proportion of NK cells in I ＋ lI stage HCC tissues was higher than Ill ＋ IV stage, especially CD3-CD56~ and CD3-CD161 ＋ NK cells. No linear relationship was found between the freauencv of NK cells subsets and zender, number of lesions. AFP. GOT.GUT, leukocytes, hepatic virus infection and TNM stage （P〉0. 05）. Conclusion The decrease of NK cells is correlated to the natural immunity and tumor progression of HCC.