世界科技研究与发展
世界科技研究與髮展
세계과기연구여발전
WORLD SCI-TECH R & D
2013年
6期
745-748
,共4页
结直肠癌%亲代细胞%肿瘤细胞球%无血清培养%上皮-间质转化
結直腸癌%親代細胞%腫瘤細胞毬%無血清培養%上皮-間質轉化
결직장암%친대세포%종류세포구%무혈청배양%상피-간질전화
colorectal cancer%monolayer cells%cancer cell spheres%serum-free medium%epithelial-mesenchymal transition
目的：探索人结直肠癌细胞系形成肿瘤细胞球的培养方法及球体内间质化标志物的表达检测。方法结直肠癌细胞系SW480、SW620、LOVO、HT29及HCT116在添加了不同细胞生长因子的无血清培养基（SFM）中悬浮培养，传代更新，诱导分化。免疫荧光技术和RT-PCR分别检测细胞球中间质化标志物N-cadherin及Vimentin的表达情况。结果5种细胞系均能在特制的SFM中形成细胞球，并稳定的传代增殖。在用血清诱导分化后，又重新贴壁生长，且与亲代细胞保持一致的细胞形态。细胞球中间质化标志物N-cadherin、Vimentin表达均上调。结论结直肠癌细胞系在低黏附及添加了细胞生长因子R-Spondin 1、Noggin的无血清环境中更容易形成悬浮生长的肿瘤细胞球，细胞球细胞比亲代细胞更具有转移侵袭能力。
目的：探索人結直腸癌細胞繫形成腫瘤細胞毬的培養方法及毬體內間質化標誌物的錶達檢測。方法結直腸癌細胞繫SW480、SW620、LOVO、HT29及HCT116在添加瞭不同細胞生長因子的無血清培養基（SFM）中懸浮培養，傳代更新，誘導分化。免疫熒光技術和RT-PCR分彆檢測細胞毬中間質化標誌物N-cadherin及Vimentin的錶達情況。結果5種細胞繫均能在特製的SFM中形成細胞毬，併穩定的傳代增殖。在用血清誘導分化後，又重新貼壁生長，且與親代細胞保持一緻的細胞形態。細胞毬中間質化標誌物N-cadherin、Vimentin錶達均上調。結論結直腸癌細胞繫在低黏附及添加瞭細胞生長因子R-Spondin 1、Noggin的無血清環境中更容易形成懸浮生長的腫瘤細胞毬，細胞毬細胞比親代細胞更具有轉移侵襲能力。
목적：탐색인결직장암세포계형성종류세포구적배양방법급구체내간질화표지물적표체검측。방법결직장암세포계SW480、SW620、LOVO、HT29급HCT116재첨가료불동세포생장인자적무혈청배양기（SFM）중현부배양，전대경신，유도분화。면역형광기술화RT-PCR분별검측세포구중간질화표지물N-cadherin급Vimentin적표체정황。결과5충세포계균능재특제적SFM중형성세포구，병은정적전대증식。재용혈청유도분화후，우중신첩벽생장，차여친대세포보지일치적세포형태。세포구중간질화표지물N-cadherin、Vimentin표체균상조。결론결직장암세포계재저점부급첨가료세포생장인자R-Spondin 1、Noggin적무혈청배경중경용역형성현부생장적종류세포구，세포구세포비친대세포경구유전이침습능력。
Objective To study on the best cultivation method for cell spheres from colorectal cancer cell lines and examine the mesenchymal markers expression in those cell spheres.Methods Colorectal cancer cell lines SW480,SW620,LOVO,HT29 and HCT116 cells were seeded in the serum free medium(SFM)supplemented with cell growth factors,the cancer cell spheres were subculture and induced to differentiate by serum supplemented medium,the expression of mesenchymal markers were investigated by immunofluorescence and RT-PCR.Results Cancer cell spheres could be generated from five colorectal cancer cell lines in SFM.These spheres were stable subculture and self-renewal, and were induced to differentiate by serum supplemented medium,these differentiated cell were adherence growth and kept the same morphol-ogy with the parental cells.These cancer cell spheres were founded expressing mesenchymal markers such as N-cadherin and Vimentin was higher than monolayer cells.Conclusion Tumor spheres can be generated and matained from colorectal cancer cell lines in SFM,and The me-tastasis ability of cell sphere was higher than monolayer cells.
