摘要
目的探讨CoCl_(2)诱导卵巢癌细胞株SKOV3产生的多倍体瘤巨细胞(PGCC)的形态及生物学特性。方法采用300μmol/L CoCl_(2)模拟缺氧环境诱导培养人卵巢癌细胞株SKOV336 h,活细胞继续常规培养、传代,20 d后收集细胞,即为PGCC组细胞;以常规培养的SKOV3细胞为对照组细胞。采用倒置显微镜观察PGCC的形成过程及形态学特点;采用免疫细胞化学法检测两组细胞肿瘤干细胞标志物OCT4、CD117的表达;采用人骨髓间质干细胞成脂诱导分化试剂盒、人骨髓间质干细胞成骨诱导分化试剂盒检测PGCC的脂肪分化及骨分化潜能;划痕实验检测PGCC的细胞迁移能力。采用1μmol/L紫杉醇分别处理PGCC组和对照组SKOV3细胞,分别在0、24、48 h用显微镜观察两组细胞形态,检测PGCC对化疗药物的抗性。结果显微镜下观察,常规培养液培养的SKOV3细胞中有极少量PGCC;CoCl_(2)可以诱导SKOV3细胞形成PGCC,其形态近似圆形、缺乏分枝,体积为SKOV3细胞的3倍及以上,细胞核多为巨核或多核,PGCC可以出芽方式产生子代细胞。免疫细胞化学染色结果显示,部分PGCC中OCT4阳性表达,无CD117阳性表达;SKOV3细胞中OCT4和CD117均未表达。使用人骨髓间质干细胞成脂诱导分化培养液培养,在第3个周期时可观察到PGCC胞质内有较大的空泡形成,油红O染色显示为橙红色、类圆形的脂滴;使用人骨髓间质干细胞成骨诱导分化培养液培养20 d,茜素红染色可观察到PGCC组细胞较对照组细胞有明显的钙结节形成。细胞划痕实验结果显示,划痕后24、48 h,使用无血清培养液培养PGCC的迁移率[(59±1)%、(66±3)%]均高于对照组细胞的迁移率[(11±3)%、(14±5)%](t值分别为32.20、19.55,均P<0.001);使用10%血清培养液培养PGCC的迁移率[(92±3)%、(100±0)%]均高于对照组细胞的迁移率[(20±6)%、(59±9)%](t值分别为16.19、8.00,均P<0.001)。1μmol/L紫杉醇处理48 h后,PGCC组细胞仍大部分存活,对照组SK
Objective To investigate the morphological and biological characteristics of polyploid tumor giant cells(PGCC)produced by ovarian cancer cell line SKOV3 induced by CoCl_(2).Methods Human ovarian cancer cell line SKOV3 was induced-cultured with 300μmol/L CoCl_(2)in the simulated hypoxic environment for 36 h,the live cells continued to be conventionally cultured and passaged,and the cells collected 20 days later were PGCC group;SKOV3 cell line cultured conventionally was the control group.The formation process and morphological characteristics of PGCC were observed by inverted microscope.The expression of tumor stem cell markers OCT4 and CD117 were detected by immunocytochemistry.The adipogenic differentiation and osteogenic differentiation potential of PGCC were detected by using human bone marrow mesenchymal stem cell adipogenic differentiation assay kit and human bone marrow mesenchymal stem cell osteogenic differentiation assay kit.The cell migration ability of PGCC was detected by scratch assay.PGCC group and control group SKOV3 cells were treated with 1μmol/L paclitaxel,and the cell morphology of the two groups was observed by microscope at 0,24 and 48 h to detect the resistance of PGCC to chemotherapy drugs.Results A small amount of PGCC was observed in SKOV3 cell line cultured in conventional medium under the microscope.CoCl_(2)can induce SKOV3 cells to form PGCC,which was nearly round in shape and lacked branching.Its volume was 3 times or more than that of SKOV3 cells,and the nuclei were usually megakaryons or multinucleates,PGCC can produce daughter cells by budding.Immunocytochemical staining showed that OCT4 was positive in some PGCC,but no CD117 was positive.Neither OCT4 nor CD117 was expressed in SKOV3 cells.When cultured with lipid-induced differentiation medium of human bone marrow mesenchymal stem cells,the formation of large vacuoles in the cytoplasm of PGCC was observed at the 3rd cycle,and orange-red,round-like lipid droplets were shown by oil red O staining.Human bone marrow mesenchymal stem
作者
乔爱琪
闫晓燕
梁钢
郗彦凤
李灵敏
Qiao Aiqi;Yan Xiaoyan;Liang Gang;Xi Yanfeng;Li Lingmin(Department of Pathology,the First Hospital of Shanxi Medical University,Taiyuan 030001,China;Department of Gynecology,the First Hospital of Shanxi Medical University,Taiyuan 030001,China;Department of Pathology,Shanxi Province Cancer Hospital,Shanxi Hospital Affiliated to Cancer Hospital,Chinese Academy of Medical Sciences,Cancer Hospital Affiliated to Shanxi Medical University,Taiyuan 030013,China;Pathological Teaching and Research Office,Shanxi Medical University,Taiyuan 030001,China)
出处
《肿瘤研究与临床》
CAS
2024年第3期199-204,共6页
Cancer Research and Clinic
关键词
卵巢肿瘤
多倍性
肿瘤干细胞
细胞分化
抗药性
Ovarian neoplasms
Polyploidy
Neoplastic stem cells
Cell differentiation
Drug resistance
