喉鳞状细胞癌干细胞的BrdU标记鉴定

Identification of squamous carcinoma stem cells in laryngocarcinoma using BrdU

目的利用5-溴脱氧尿嘧啶核苷(bromode-oxyuridine,BrdU)标记滞留细胞的方法,研究小鼠喉黏膜和喉鳞状细胞癌(简称喉鳞癌)实体瘤内可能存在的干细胞及其分布特点。方法出生3 d的昆明小鼠,随机分为实验组和对照组。实验组小鼠皮下注射BrdU,对照组皮下注射生理盐水。第2、4、8周分别处死6只小鼠,并取喉黏膜进行免疫组化检测,计算标记滞留细胞的阳性率。被BrdU标记的喉鳞癌Hep细胞注入到6只裸鼠皮下成瘤,8周后处死并取肿瘤,HE染色及免疫组化检测。结果实验组标记后2、4、8周,小鼠鳞状上皮和腺上皮均有标记滞留细胞表达,对照组标记后无标记滞留细胞表达。不同时间点标记滞留细胞阳性率差异具有显著性(F=140.319,P<0.05)。鳞状上皮基底层中标记滞留细胞阳性率显著高于腺体细胞(t分别为6.376、10.944、15.376,P<0.05)。结论小鼠喉黏膜和喉鳞癌实体瘤中存在标记滞留细胞,这些标记滞留细胞被认为是喉黏膜成体干细胞及喉鳞癌肿瘤干细胞。

OBJECTIVE To identify the existence of stem cells and its distribution in mouse laryngeal mucosa and Laryngeal squamous cell carcinoma by using the label–retaining cells with bromodeoxyurdine（BrdU）.METHODS Thirty six Kunming white mice 3 days after born were randomly divided into two groups.BrdU was subcutaneous injected in experiment group and NS in control group.The 6 mice in each group were killed after injected 2w,4w and 8w.The laryngeal mucosa was fixed for immunohistochemistry and the percentages of label-retaining cells were calculated.A Hep-2 cell line of laryngocarcinoma was labeled with BrdU in vitro and then engrafted into the back of 6 nude mice.The nude mice were killed after injected 8w.The tumors were fixed for immunohistochemistry and pathology.RESULTS The label–retaining cells were found in squamous cell epithelium and gland epithelium at 2w,4w,8w in experimental group.There were no label–retaining cells in control group.The percentages of label–retaining cells in different time were different significantly（F=140.319,P0.05）.The percentages of label–retaining cells in basement membrane of squamous cell epithelium were higher than that in gland epithelium at each time（t=6.376、10.944、15.376,P0.05）.Label–retaining cells were detected in laryngocarcinoma after 8w in nude mice.CONCLUSIONThere are label–retaining cells in the mouse laryngeal mucosa and laryngocarcinoma.It is possible that stem cells exist in these label–retaining cells.