CAJ | 학술논문

目的：利用紫杉醇联合无血清培养完成对 A549肺腺癌始动细胞的富集并鉴定富集亚群的干细胞特性。方法对数生长期的 A549细胞经胰酶消化，干细胞培养基重悬，得到成球状生长的细胞；传至第2代时加入紫杉醇作用48 h ，离心去除死细胞和紫杉醇，换新鲜干细胞培养基培养，至存活细胞恢复克隆生长后鉴定其干细胞相关特性。结果紫杉醇联合无血清培养方式成功从 A549细胞中富集得到肿瘤干细胞，该群细胞高表达分化抗原簇蛋白133/人上皮细胞黏附分子（CD133/CD326），具有多向分化潜能、高表达干细胞相关基因及更强的致瘤能力，具备干细胞相关特性。结论紫杉醇联合无血清培养富集得到高表达CD133/CD326分子的细胞亚群，该亚群具备干细胞相关特性，可用于后续肺腺癌干细胞生物学特性的研究。
목적：이용자삼순연합무혈청배양완성대 A549폐선암시동세포적부집병감정부집아군적간세포특성。방법대수생장기적 A549세포경이매소화，간세포배양기중현，득도성구상생장적세포；전지제2대시가입자삼순작용48 h ，리심거제사세포화자삼순，환신선간세포배양기배양，지존활세포회복극륭생장후감정기간세포상관특성。결과자삼순연합무혈청배양방식성공종 A549세포중부집득도종류간세포，해군세포고표체분화항원족단백133/인상피세포점부분자（CD133/CD326），구유다향분화잠능、고표체간세포상관기인급경강적치류능력，구비간세포상관특성。결론자삼순연합무혈청배양부집득도고표체CD133/CD326분자적세포아군，해아군구비간세포상관특성，가용우후속폐선암간세포생물학특성적연구。
Objective To obtain the lung adenocarcinoma initiating cells from the A 549 cell line based on paclitaxel treatment combination with serum-free cultivation and to validate spared cells can represent tumor initiating cells (TICs) .Methods After dis-sociated by trypsogen ,about 106 /mL cells were suspended in serum-free medium supplemented with 0 .4% bovine serum albumin (BSA) ,insulin ,basic fibroblast growth factor (bFGF) ,human recombinant epidermal growth factor (EGF) and obtained spheroid cells .At the second passage ,paclitaxel was added at a concentration of 100 nmol/L for 48 h and then replaced with completely fresh medium once or twice per week until new spheroids emerged .Results The subpopulation of cells that survived serum-free cultiva-tion and paclitaxel treatment could highly express the cluster of differentiation 133/cluster of differentiation (CD133/CD326) mo-lecular markers and have features of stemness including differentiation ,high expression of cancer stem cells (CSCs)-associated genes and stronger capability of tumorigenesis .Conclusion The survived subpopulation that highly express the CD 133/CD326 molecu-lar markers presenting the characteristics of stemness in vitro and in vivo ,and could be used in future researches of biological functions .