摘要
目的 :探讨甲状腺未分化癌耐药和转移中肿瘤干细胞(cancer stem cells,CSCs)和上皮细胞间质转化(epithelialmesenchymal transition,EMT)的作用。方法 :以甲状腺未分化癌细胞株SW1736为研究对象,采用流式荧光激活分选技术从SW1736细胞株中分选出具有干细胞特性的侧群(side population,SP)细胞。用阿霉素诱导非侧群(non-SP)细胞耐药。在SP、nonSP及non-SP耐药细胞中,采用体外克隆形成试验检测克隆形成能力;四甲基偶氮唑蓝(MTT)法检测阿霉素对增殖的影响;realtime PCR比较干细胞标志nestin、ABCG2,肿瘤耐药基因MDR1,EMT标记物E-cadherin、β-catenin、vimentin、slug、N-cadherin表达情况。结果:SW1736细胞中存在0.8%左右的SP细胞,SP细胞克隆形成率明显高于non-SP细胞。non-SP耐药细胞的半数抑制浓度明显高于non-SP细胞,且non-SP耐药细胞流式分选不能获得SP细胞,只能得到non-SP细胞。real-time PCR结果显示,SP较non-SP细胞高表达nestin、ABCG2、MDR1、N-cadherin、β-catenin、vimentin及slug;non-SP耐药细胞较non-SP细胞MDR1、slug表达上调;均未检出E-cadherin。结论 :阿霉素诱导耐药不能将非肿瘤干细胞(non-SP细胞)转化为获得性肿瘤干细胞(SP细胞),但可导致EMT的异常激活。SP细胞可能是肿瘤耐药、复发、转移的根源。
Objective：To explore the role of cancer stem cells（CSCs） and epithelial-mesenchymal transition（EMT） in drug resistance and metastasis of anaplastic thyroid carcinoma. Methods：Human anaplastic thyroid cancer cell line,SW1736,was sorted for side population（SP） cells which had stem cell characteristics by Fluorescent Activated Cell Sorting（FACS）. Non-SP cells were treated with doxorubicin to establish drug resistant cell modes. Among SP,non-SP and non-SP drug resistant cells,the clonal formation assay was performed to evaluate the self-renewal potential;3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT） cell proliferation assay was adopted to examine effects of doxorubicin on the proliferation;Genes expression of stem cell markers-nestin and ATP-binding cassette superfamily G member 2（ABCG2）,the gene related to cancer resisitance and relapse-multidrug resistance gene（MDR1） and some EMT associated genes-E-Cadherin,β-catenin,vimentin,Slug and N-cadherin,were compared by performing real-time PCR.Results：SW1736 line cells contend 0.8% side population cells. Clonal formation assay revealed that SP cells displayed markedly higher clonogenic potential than non-SP cells. MTT cell proliferation assay showed that the half maximal inhibitory concentration（IC50） of nonSP drug resistant cells was obviously higher than that of non-SP cells. SP cells displayed higher genes expression of nestin,ABCG2,MDR1,slug,β-catenin,vimentin and N-cadherin,compared with non-SP cells. The gene expressions of slug and MDR1 were higher in non-SP drug resistant cells than those of non-SP cells. E-cadherin was not detected in all cell types. Conclusion： Doxorubicin can＇t make the transition of non-CSCs（namely non-SP） to acquired CSCs（namely SP）,but can induce the aberrant activation of EMT. SP cells may have significant impact on tumor drug resistance,recurrence and metastasis.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2014年第12期1627-1631,1637,共6页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家自然科学基金资助(81102032)
关键词
甲状腺
肿瘤干细胞
侧群细胞
上皮细胞间质转化
耐药
thyroid
cancer stem cells
side population
epithelial-mesenchymal transition
drug resistance
