世界科技研究与发展
世界科技研究與髮展
세계과기연구여발전
WORLD SCI-TECH R & D
2014年
6期
660-663
,共4页
结直肠癌%肿瘤干细胞%干细胞标志物%无血清培养%CD1 33
結直腸癌%腫瘤榦細胞%榦細胞標誌物%無血清培養%CD1 33
결직장암%종류간세포%간세포표지물%무혈청배양%CD1 33
colorectal carcinoma cell lines%cancer stem/tumor-initiating cells%stem cell related marker%serum-free cul-ture%CD1 33
目的：探讨人结直肠肿瘤干细胞在体外分化过程中细胞形态和干细胞相关标志物CD133的表达变化，为进一步研究结直肠肿瘤干细胞分化走向提供实验依据。方法取来源于人结直肠癌的细胞系HCT116，无血清培养分离出CD133＋细胞，加血清诱导分化，相差显微镜下观察其形态变化；在未分化状态下无血清培养第7天和14天与血清诱导分化后收集细胞，利用流式细胞仪检测干细胞标志物CD133的表达量，采用激光共聚焦检测CD133表面标记分子的表达。结果1）细胞形态：无血清培养分离的CD133＋细胞，在生长过程中聚集成规则的细胞球，血清诱导后即贴壁生长，贴壁形态与同来源细胞形态一致，且再次无血清悬浮培养后聚集成球稳定生长。2）标志物变化：结直肠肿瘤干细胞未分化时CD133表面标记分子高表达，流式细胞仪检测未分化细胞 CD133第7天表达率为（20．4±0．52）％，第14天表达率为（78．5±2．80）％，分化后表达率为（0．50±0．17）％。结论细胞形态和标志物表达改变均表明高表达CD133＋的HCT116结直肠癌肿瘤干细胞可定向分化为同源的结直肠癌细胞，CD133＋细胞经血清诱导后表达下调而使细胞失去干细胞特性。
目的：探討人結直腸腫瘤榦細胞在體外分化過程中細胞形態和榦細胞相關標誌物CD133的錶達變化，為進一步研究結直腸腫瘤榦細胞分化走嚮提供實驗依據。方法取來源于人結直腸癌的細胞繫HCT116，無血清培養分離齣CD133＋細胞，加血清誘導分化，相差顯微鏡下觀察其形態變化；在未分化狀態下無血清培養第7天和14天與血清誘導分化後收集細胞，利用流式細胞儀檢測榦細胞標誌物CD133的錶達量，採用激光共聚焦檢測CD133錶麵標記分子的錶達。結果1）細胞形態：無血清培養分離的CD133＋細胞，在生長過程中聚集成規則的細胞毬，血清誘導後即貼壁生長，貼壁形態與同來源細胞形態一緻，且再次無血清懸浮培養後聚集成毬穩定生長。2）標誌物變化：結直腸腫瘤榦細胞未分化時CD133錶麵標記分子高錶達，流式細胞儀檢測未分化細胞 CD133第7天錶達率為（20．4±0．52）％，第14天錶達率為（78．5±2．80）％，分化後錶達率為（0．50±0．17）％。結論細胞形態和標誌物錶達改變均錶明高錶達CD133＋的HCT116結直腸癌腫瘤榦細胞可定嚮分化為同源的結直腸癌細胞，CD133＋細胞經血清誘導後錶達下調而使細胞失去榦細胞特性。
목적：탐토인결직장종류간세포재체외분화과정중세포형태화간세포상관표지물CD133적표체변화，위진일보연구결직장종류간세포분화주향제공실험의거。방법취래원우인결직장암적세포계HCT116，무혈청배양분리출CD133＋세포，가혈청유도분화，상차현미경하관찰기형태변화；재미분화상태하무혈청배양제7천화14천여혈청유도분화후수집세포，이용류식세포의검측간세포표지물CD133적표체량，채용격광공취초검측CD133표면표기분자적표체。결과1）세포형태：무혈청배양분리적CD133＋세포，재생장과정중취집성규칙적세포구，혈청유도후즉첩벽생장，첩벽형태여동래원세포형태일치，차재차무혈청현부배양후취집성구은정생장。2）표지물변화：결직장종류간세포미분화시CD133표면표기분자고표체，류식세포의검측미분화세포 CD133제7천표체솔위（20．4±0．52）％，제14천표체솔위（78．5±2．80）％，분화후표체솔위（0．50±0．17）％。결론세포형태화표지물표체개변균표명고표체CD133＋적HCT116결직장암종류간세포가정향분화위동원적결직장암세포，CD133＋세포경혈청유도후표체하조이사세포실거간세포특성。
Objective To pursue the changes of cell morphology,expression of stem cell related markers of colorectal cancer stem cells after differentiation in vitro.Methods Tumor stem cells of the line CD1 33 +were obtained from colorectal cancer cell line HCT1 1 6 cells were seeded in the serum free medium(SFM)supplemented with cell growth factors.Then cultured to differentiate in medium containing 1 0% fetal bovine serum.The morphology of the cells was observed under phase con-trast microscope.Cells were collected respectively before differentiation and 7,1 4 days after the differentiation.The cell sur-face markers such as CD1 33 was investigated by flow cytometry.Results 1 )HCT1 1 6 cells were formed cancer cell spheres in SFM,Sphere cells are able to produce a heterogeneous cell population that can produce spheres.Cancer cell spheres were induced to differentiate by serum supplemented medium,these differentiated cell were adherence growth and kept the same morphology with the parental cells.2)These cancer cell spheres were founded expressing markers such as CD1 33 was higher than differentiated cells.The expression rates of CD1 33 was (20. 4 ±0. 52)% in the 7 days ,(78. 5 ±2. 80)% in the 1 4 days,and (0. 50 ±0. 1 7)% of the differentiated cells(P<0. 05)Conclusion The changes of morphology and cell marker expression in the high-expression CD1 33 +CRC cells were oriented differentiated into the same origin CRC cells,and the CD1 33 +cells were losing stem cell ability through serum-induced.
