肿瘤相关巨噬细胞浸润在垂体腺瘤卒中发生中的作用及相关性

Tumor-associated macrophages infiltration in pituitary adenoma apoplexy

目的探讨肿瘤相关巨噬细胞浸润在垂体腺瘤卒中发生中的作用和相关性。方法筛选获得华中科技大学同济医学院附属同济医院神经外科经影像资料、术中和病理结果证实垂体腺瘤或垂体腺瘤卒中患者病例样本67例,采用苏木精-伊红(HE)染色、免疫组织化学(IHC)染色的方法检测样本中免疫细胞的分布,分析其浸润水平与临床特征(性别、年龄、激素类型、侵袭性、肿瘤大小)的相关性。在体外细胞实验中,运用膜联蛋白V-异硫氰酸荧光素(Annexin V-FITC)/碘化丙锭(PI)双染色的流式细胞技术,评估被垂体瘤细胞系(GH3、AtT20、TtT/GF)培养基活化的巨噬细胞对垂体瘤细胞系的影响。结果HE染色结果显示20.9%的病例样本组织(卒中组13/39,非卒中组1/28)有广泛干酪样坏死伴纤维素样组织沉积。Pearson’s相关分析显示离子钙接头蛋白分子-1(Iba-1)染色强度与卒中相关(P<0.01),与性别(P>0.05)、年龄(R^(2)=0.011,P>0.05)、激素类型(P>0.05)、侵袭性(P>0.05)、肿瘤大小(R^(2)=0.009,P>0.05)无相关。在体外细胞培养实验中发现被垂体瘤细胞系(GH3、AtT20、TtT/GF)培养基刺激的巨噬细胞系(Raw246.7)会促进垂体瘤细胞系的凋亡[GH3:(24.39±0.860)%比(6.467±0.709)%,t=20.56,P<0.01;AtT20:(20.87±1.896)%比(7.403±0.261)%,t=10.94,P<0.01;TtT/GF:(27.970±1.399)%比(8.563±0.632)%,t=26.20,P<0.01]。结论垂体腺瘤卒中肿瘤组织相较于非卒中肿瘤组织存在更高水平的肿瘤相关巨噬细胞浸润,活化的巨噬细胞可能通过促进垂体腺瘤细胞凋亡的方式促进垂体腺瘤的出血转化。

Objective To explore the relationship between tumor-associated macrophages infiltration and the occurrence of pituitary adenoma apoplexy(PAA).Methods Tissues of 67 PAA patients were obtained from the Department of Neurosurgery,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology.The diagnosis was confirmed by magnetic resonance imaging,histopathology,and intraoperative evidence.The morphological changes of tissue and cells were observed through hematoxylin and eosin(HE)staining.Infiltration level of macrophages marked by Ionized calcium binding adaptor molecule-1(Iba-1)in PAA specimens was detected by immunohistochemistry.Correlation between infiltration level of macrophages in pituitary specimens and the occurrence of PAA,clinical characteristics(gender,age,hormone type,invasion,size)was analyzed.Flow cytometry with annexin V-fluoresceine isothiocyanate(Annexin V-FITC)/propidium iodide(PI)double staining was used to evaluate the apoptosis level of pituitary adenoma cells cultured with macrophages.Results Immunohistochemical staining showed that 20.9%of the specimens(13/39 in the PAA group and 1/28 in the non-apoplexy group)had extensive caseous necrosis with fibrous tissue deposition.Pearson’s correlation analysis showed that the intensity of Iba-1 staining was related to apoplexy(P<0.01),and had no significant correlation with hormone type(P>0.05),aggressiveness(P>0.05),gender(P>0.05),tumor size(R^(2)=0.009,P>0.05),and age(R^(2)=0.011,P>0.05).Macrophages activated by culture medium of pituitary tumor cell lines(GH3,AtT20,TtT/GF)promoted the apoptosis level of cell lines[GH3:(24.39±0.860)%vs.(6.467±0.709)%,t=20.56,P<0.01;AtT20:(20.87±1.896)%vs.(7.403±0.261)%,t=10.94,P<0.01;TtT/GF:(27.970±1.399)%vs.(8.563±0.632)%,t=26.20,P<0.01].Conclusion Pituitary adenomas with apoplexy have higher level of tumor-associated macrophages infiltration than non-apoplexy.Activated macrophages can promote the apoptosis of pituitary adenoma cells.