一例树鼩毛发上皮瘤的诊断及细胞生物学特性观察

doi:10.12300/j.issn.1674-5817.2022.173

实验动物与比较医学 ›› 2023, Vol. 43 ›› Issue (4): 440-445.DOI: 10.12300/j.issn.1674-5817.2022.173

一例树鼩毛发上皮瘤的诊断及细胞生物学特性观察

翟珊珊¹(), 梁亮¹, 曹颖颖¹, 李竹欣¹, 王青¹, 陶俊宇¹^,², 运晨霞¹^,³, 冷静¹^,²^,³()(), 唐海波¹^,²()()

^1.广西中医药大学, 南宁 530200
^2.广西高发传染病中西医结合转化医学重点实验室, 南宁 530200
^3.广西特色实验动物模型重点实验室, 南宁 530200

收稿日期:2022-11-08 修回日期:2023-04-20 出版日期:2023-08-25 发布日期:2023-09-14
通讯作者: 冷静（1963—），女，博士，教授，研究方向：天然免疫。E-mail： lj986771558@163.com。ORCID： 0000-0001-6115-2153；
唐海波（1983—），男，博士，副研究员，研究方向：抗感染免疫。E-mail: thb514@163.com。ORCID: 0000-0002-1815-699X
作者简介:翟珊珊（1996—），女，硕士，主要从事抗感染免疫研究。E-mail： zhaishanshan369@126.com
基金资助:
广西自然科学基金项目“基于树鼩模型探索TLR8在狂犬病发病过程中的作用机理”(2020GXNSFAA297188);广西高等学校千名中青年骨干教师培育计划“广西中医药大学-唐海波”（第三期）(桂教师范（2019）81号);广西中医药大学高层次人才培育创新团队“冷静团队”(2022B006);2023年广西中医药多学科交叉创新团队项目“中医药生物新材料关键技术研究与智能装备开发学科交叉创新团队”(GZKJ2303)

Diagnosis of Trichoepithelioma in a Tree Shrew and Observation of Cell Biological Characteristics

Shanshan ZHAI¹(), Liang LIANG¹, Yingying CAO¹, Zhuxin LI¹, Qing WANG¹, Junyu TAO¹^,², Chenxia YUN¹^,³, Jing LENG¹^,²^,³()(), Haibo TANG¹^,²()()

^1.Guangxi University of Chinese Medicine, Nanning 530200, China
^2.Guangxi Key Laboratory of Translational Medicine for Treating High-Incidence Infectious Diseases with Integrative Medicine, Nanning 530200, China
^3.Key Laboratory of Characteristic Experimental Animal Models of Guangxi, Nanning 530200, China

Received:2022-11-08 Revised:2023-04-20 Published:2023-08-25 Online:2023-09-14
Contact: LENG Jing (ORCID: 0000-0001-6115-2153), E-mail: lj986771558@163.com;
TANG Haibo (ORCID: 0000-0002-1815-699X), E-mail: thb514@163.com

摘要/Abstract

摘要：

目的观察实验动物树鼩的原发性肿瘤，为研究毛发上皮瘤的发生机制及防治措施提供基础。方法将常规自然饲养过程中发现胸腹部长有肿块的1例树鼩麻醉后，手术摘除肿块，肿块组织经石蜡切片后行HE染色及免疫组织化学染色，并对肿瘤细胞进行分离和传代培养。分离获得的肿瘤细胞接种至健康树鼩及裸小鼠皮下。每天观察1次肿瘤细胞的体内成瘤情况，裸小鼠连续观察2个月，树鼩观察6个月以上。结果 HE染色显示发病树鼩腹部肿块组织中真皮内基底样细胞整体排列似串花瓣样，形成巢状及条索状且边缘界限清晰的肿瘤；进一步放大倍数后观察发现肿瘤细胞呈栅栏状、基底状排列，核染色深，细胞质少。免疫组织化学染色显示，肿瘤细胞中CK蛋白强阳性表达，ki-67蛋白呈低比例阳性表达；肿瘤细胞间质中Vimentin强阳性表达，Bcl2和CD10呈低比例阳性表达。分离获得的肿瘤细胞在含10%胎牛血清的DMEM培养液中生长良好，可传代培养。健康树鼩及裸小鼠皮下接种肿瘤细胞后，均未见肿瘤形成。结论结合肿瘤发生部位、大体形态结构、病理切片HE染色及免疫组织化学结果，综合诊断该例树鼩胸腹部肿块为毛发上皮细胞瘤。

关键词: 树鼩, 毛发上皮瘤, 诊断, 肿瘤细胞移植, 裸小鼠

Abstract:

Objective To observe the primary tumor of tree shrews and to provide a basis for studying the pathogenesis and prevention of trichoepithelioma. Methods A tumor was discovered in the chest and abdomen of a tree shrew during natural cultivation. The tree shrew was anesthetized, and the tumor was surgically removed. Hematoxylin and eosin (HE) staining and immunohistochemical staining were performed on the tumor tissue after paraffin section, and the tumor cells were isolated and cultured by passage. The isolated tumor cells were subcutaneously injected into healthy tree shrews and nude mice. The tumorigenesis of tumor cells in vivo was observed once a day, with nude mice continuously observed for 2 months and tree shrews observed for more than 6 months. Results HE staining showed that the basal cells in the dermis were arranged as a whole, like a string of petals, forming nests and stripe-like structures with clear boundaries. The observation results after magnification revealed that the tumor cells were arranged in a pallisade-like and basal pattern, with deep nuclear staining and minimal cytoplasmic. Immunohistochemical staining showed the high expression of CK protein and low proportion expression of ki-67 protein in tumor cells, as well as the high expression of vimentin and low expressions of Bcl2 and CD10 in tumor cell mesenchyme. The isolated tumor cells grew well in DMEM medium containing 10% fetal bovine serum and could be cultured by passage, but no tumor formation was observed in healthy tree shrews and nude mice inoculated with tumor cells. Conclusion Combined with the location of the tumor, overall morphology, HE staining, and immunohistochemical results, the thoracoabdominal mass of the tree shrew was diagnosed as a trichoepithelioma.

Key words: Tree shrew, Trichoepithelioma, Diagnosis, Tumor cell transplantation, Nude mice

中图分类号:

R-332

翟珊珊, 梁亮, 曹颖颖, 李竹欣, 王青, 陶俊宇, 运晨霞, 冷静, 唐海波. 一例树鼩毛发上皮瘤的诊断及细胞生物学特性观察[J]. 实验动物与比较医学, 2023, 43(4): 440-445.

Shanshan ZHAI, Liang LIANG, Yingying CAO, Zhuxin LI, Qing WANG, Junyu TAO, Chenxia YUN, Jing LENG, Haibo TANG. Diagnosis of Trichoepithelioma in a Tree Shrew and Observation of Cell Biological Characteristics[J]. Laboratory Animal and Comparative Medicine, 2023, 43(4): 440-445.

图/表 3

图1 树鼩原发肿瘤组织病理学变化（HE染色）注：Ａ，50倍视野（比例尺大小为200 μm）下，肿块组织边缘轮廓清晰，细胞呈条索状、栅栏状排列；Ｂ，200倍视野（比例尺大小为50 μm）下，肿瘤细胞呈基底细胞样，核深染，胞质少。

Figure 1 Histopathological changes of primary tumor in the tree shrews （HE staining）Note： A， The edge of the tumor tissue was clearly outlined and the tumor cells were arranged in a striated and palisade pattern （×50）； B， HE-stained tumor cells were basal cell-like with deep nuclear staining and minimal cytoplasm （×200）.

图2 树鼩毛发上皮瘤的免疫组织化学染色结果（DAB染色，×100）注：A，肿瘤细胞中CK强阳性表达（红色箭头所示）；B，肿瘤细胞间质中vimentin呈强阳性表达（红色箭头所示）；C，肿瘤细胞中ki-67低指数阳性表达（红色箭头所示）；D，肿瘤细胞间质中Bcl2呈低比例阳性表达（红色箭头所示）；E，肿瘤细胞间质中CD10呈低比例阳性表达（红色箭头所示）；F，肿瘤细胞中CD34表达呈阴性。

Figure 2 Immunohistochemistry of trichoepithelioma from a tree shrew（DAB staining，×100）Note： A， Strongly positive expression of CK in tumor cells （As indicated by the red arrow）； B， Strongly positive expression of vimentin in tumor cell mesenchyme （as indicated by the red arrow）； C， Low index expression of ki-67 in tumor cells （as indicated by the red arrow）； D， Low positive expression of Bcl2 in tumor cell mesenchyme （as indicated by the red arrow）； E， Low proportion expression of CD10 in tumor cell mesenchyme （as indicated by the red arrow）； F， Negative expression of CD34 in tumor cells.

图3 树鼩自发性毛发上皮瘤原代肿瘤细胞分离培养及形态特征观察注：A～C，肿瘤细胞分离培养后第1天、第2天、第5天（×100）。D，肿瘤细胞呈多形性、不规则、有触角（×400）。

Figure 3 Culture of primary tumor cells from a tree shrew trichoepithelioma and morphological observationNote：A， Day 1 after tumor cell isolation and culture （×100）； B， Day 2 after tumor cell isolation and culture （×100）； C， Day 5 after tumor cell isolation and culture （×100）. D， Tumor cells were pleomorphic， irregular， and tentacled （×400）.

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一例树鼩毛发上皮瘤的诊断及细胞生物学特性观察

Diagnosis of Trichoepithelioma in a Tree Shrew and Observation of Cell Biological Characteristics

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