鼻咽癌干细胞标志物的检测意义

Significance of Detection of Nasopharyngeal Cancer Stem Cell Markers

目的检测鼻咽癌干细胞可能标志物,为鼻咽癌发生机制的研究提供依据。方法采用免疫组化法,检测在鼻咽癌、鼻咽黏膜慢性炎中CD133和CK19表达及鼻咽癌细胞株裸鼠移植瘤中CK19表达;采用免疫细胞化学法检测CD133和CK19在鼻咽癌或永生化细胞株NP69中的表达;PCR法检测鼻咽癌细胞株中CD133mRNA表达情况。结果CK19在大部分鼻咽癌组织中呈强阳性表达(37/39,67.2%),黏膜慢性炎柱状上皮及化生鳞状上皮中均呈强阳性表达。CK19在5-8F、6-10B和CNE2裸鼠移植瘤中都有散在点巢状阳性,但在细胞株中呈阴性表达。39例鼻咽癌组织中8例仅有个别癌巢的边缘见散在CD133+细胞,2例黏膜上皮中主要是基底部细胞呈阳性。11例鼻咽黏膜组织中均未见CD133阳性细胞。PCR检测结果显示CD133基因在3种鼻咽癌细胞株中均有清晰的条带。结论 CK19在鼻咽癌组织和移植瘤中呈强表达及在细胞株中呈罕见表达的差异型,可能与微环境有关,而非鼻咽癌干细胞标志物。CD133在鼻咽癌肿瘤边缘组织中稀少表达,且在细胞株中检测出其RNA表达,可能为鼻咽癌干细胞标志物。

Objective To detect possible stem cell markers of nasopharyngeal carcinoma ( NPC) ,and to provide basis for carcinogenesis of nasopharyngeal carcinoma .Methods CD133 and CK19 were detected by immunohistochemistry in specimens of nasopharyngeal carcinoma ,chronic nasopharyngitis ,xenograft tumors of NPC cell lines ,cancer cell lines and immortalized cell line NP69.CD133 mRNA was detected by PCR in NPC cell lines .Results CK19 was strongly expressed in most of NPC cancer cells (37/39,67.2%),columnar epithelial and metaplastic squamous epithelium of chronic nasopharyngitis .CK19 expression al-most could not be found in cell lines 5-8F,6-10B and CNE2 while in xenograft tumors these cancer cell lines were positively stained with CK19.Several CD133 positive cells could be seen in normal epithelial cells and NP 69.But positive cells could not be found in 5-8F,6-10B and CNE2.Gel electrophoresis of PCR amplification with CD 133 showed that 5-8F and 6-10B and CNE2 had distinct straps .Conclusion CK19 is not the biomarker of nasopharyngeal stem cells;and the expression model of CD 133 in NPC tissues and cell lines suggest that CD 133 maybe a potential marker of nasopharyngeal stem cells .