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<0.05) . 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 inⅠ+Ⅱstage HCC tissues was higher thanⅢ+Ⅳstage, especially CD3-CD56+and CD3-CD161+NK cells. No linear relationship was found between the frequency of NK cells subsets and gender, 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.%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，并收集例健康志愿者的外周血作为对照。免疫组化法检测例HCC癌组织和癌旁组织及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细胞比例分别为（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。
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