CAJ | 학술논문

目的：观察生殖干细胞自我更新关键调节基因Piwil2重编程人包皮成纤维细胞获得的肿瘤干细胞（ Piwil2?iCSC）体内成瘤组织的组织学特征。方法挑选Piwil2?iCSC肿瘤球样克隆细胞进行裸鼠体内成瘤实验，组织学观察Piwil2?iCSC肿瘤组织病理特点，逆转录聚合酶链反应（ RT?PCR）和免疫组化法检测Piwil2?iCSC肿瘤组织中干细胞标志物和常见三胚层肿瘤标志物，免疫组化法检测Piwil2?iCSC肿瘤组织中生殖细胞肿瘤标志物。结果接种2周时，裸鼠体内皮下100％形成肿瘤。Piwil2?iCSC肿瘤组织DAPI染色显示，Piwil2?GFP 标记定位细胞核，且满视野高密度表达。 HE染色显示，肿瘤细胞核分裂象高，每高倍镜下＞10个核分裂象，核增大畸形。 RT?PCR检测显示，Piwil2?iCSC肿瘤组织仍表达Piwil2、部分干细胞自我更新标志物和三胚层特征标志物。免疫组化染色显示， Piwil2?iCSC肿瘤组织表达相关干细胞标志物、三胚层多种肿瘤标志物和生殖细胞肿瘤标志物AFP、HCG。结论 Piwil2?iCSC肿瘤可能为胚胎性未分化小细胞癌，以未成熟畸胎瘤混有卵黄囊瘤和绒癌成分可能性大，可为研究相关肿瘤的肿瘤干细胞起源或发生机制及诊疗方法提供研究模型。
목적：관찰생식간세포자아경신관건조절기인Piwil2중편정인포피성섬유세포획득적종류간세포（ Piwil2?iCSC）체내성류조직적조직학특정。방법도선Piwil2?iCSC종류구양극륭세포진행라서체내성류실험，조직학관찰Piwil2?iCSC종류조직병리특점，역전록취합매련반응（ RT?PCR）화면역조화법검측Piwil2?iCSC종류조직중간세포표지물화상견삼배층종류표지물，면역조화법검측Piwil2?iCSC종류조직중생식세포종류표지물。결과접충2주시，라서체내피하100％형성종류。Piwil2?iCSC종류조직DAPI염색현시，Piwil2?GFP 표기정위세포핵，차만시야고밀도표체。 HE염색현시，종류세포핵분렬상고，매고배경하＞10개핵분렬상，핵증대기형。 RT?PCR검측현시，Piwil2?iCSC종류조직잉표체Piwil2、부분간세포자아경신표지물화삼배층특정표지물。면역조화염색현시， Piwil2?iCSC종류조직표체상관간세포표지물、삼배층다충종류표지물화생식세포종류표지물AFP、HCG。결론 Piwil2?iCSC종류가능위배태성미분화소세포암，이미성숙기태류혼유란황낭류화융암성분가능성대，가위연구상관종류적종류간세포기원혹발생궤제급진료방법제공연구모형。
Objective To observe the histological features of tumor?bearing tissues formed by human fibroblasts after reprograming by spermatogonial stem cell self?renewal key regulating gene Piwil2 ( Piwil2?iCSC) . Methods Piwil2?iCSC tumor spheroids?like colonies were selected for tumor formation assay in four nude mice. Pathological features of Piwil2?iCSC tumors were observed by histology. Stem cell markers and common triploblastic markers were detected by reverse transcriptase?polymerase chain reaction ( RT?PCR ) assay and immunohistochemistry. Germ cell tumor markers were detected by immunohistochemical examination. Results Two weeks after inoculation, subcutaneous tumors were formed in all the four nude mice with a tumor formation rate of 100%. In the Piwil2?iCSC tumor tissues, Piwil2?GFP+ cells showed high?density nuclear expression and were widely observed in DAPI?stained sections. Numerous mitotic figure of the neoplastic cells were seen (> 10 cells/field of vision under high magnification) in HE?stained sections. Enlarged abnormal cell nuclei were observed. RT?PCR assay showed that Piwil2?iCSC tumors still expressed Piwil2 and some self?renewal and pluripotent markers of stem cells and some markers of triploblastic differentiation. Immunohistochemical staining showed that the tumors expressed stem cell markers, triploblastic markers and germ cell tumor markers AFP and HCG. Conclusions Piwil2?iCSC tumors are probably undifferentiated embryonic small cell carcinoma, most likely to be immature teratoma, mixed with yolk sac tumor and choriocarcinoma components. It can be used as a useful model for the research of origin or genesis mechanism of cancer stem cells and the treatment of relevant tumors.