摘要
目的 分选肝癌细胞株SMMC-7721中的侧群（SP）细胞,探讨其体外侵袭性与上皮-间充质转化（EMT）的关系.方法 采用流式细胞荧光激活分选（FACS）技术将SMMC-7721细胞分为SP细胞和非侧群（NSP）细胞两个亚群,以Transwell法检测SP细胞和NSP细胞侵袭能力,实时定量逆转录聚合酶链反应（RT-qPCR）和Western blot技术对两个亚群细胞波形蛋白（Vimentin）、E-钙黏蛋白（E-cadherin）、N-钙黏蛋白（N-cadherin）和Snail的表达进行分析.结果 Transwell侵袭实验中SP细胞穿膜细胞数（66.3±7.4）个明显高于NSP细胞穿膜细胞数（27.5±6.5）个（P＜0.01）；SP细胞Vimentin、N-cadherin和Snail mRNA的表达水平分别是NSP细胞的9.527、6.834、8.173倍,NSP细胞E-cadherin mRNA的表达水平是SP细胞的5.353倍,差异均有统计学意义（P＜0.01）；E-cadherin、Vimentin、N-cadherin和Snail蛋白SP细胞和NSP细胞中的表达差异有统计学意义（0.174±0.014和0.935±0.012、1.117±0.012和0.314±0.011、0.975±0.017和0.179±0.013、0.917±0.014和0.202±0.013,P＜0.01）.结论 肝癌SMMC-7721细胞中SP细胞的体外高侵袭性可能与EMT有关.
Objective To study the in vitro invasiveness of side population （SP） cells purified from hepatoma SMMC-7721 cells and the expression of epithelial-mesenchymal transition （EMT） markers of SP cells and non-SP （NSP） cells,and to explore the relationship between invasiveness of SP cells and EMT.Methods Fluorescence-activated cell sorting （FACS） was used to sort SP cells and non-SP （NSP） cells from SMMC-7721 cell line.The in vitro invasiveness of both subpopulations was tested by using Transwell chamber assay.Quantitative real-time reverse transcriptase-pelymerase chain reaction （RT-qPCR）and Western blotting were applied to evaluate the expression of Vimentin,E-cadherin,N-cadherin and Snail in SP cells and NSP cells,respectively.Results Transwell invasion assay revealed the number of SP cells （66.3 ±7.4） penetrating the basilar membrane was significantly greater than that of NSP cells （27.5 ±6.5）（P 〈0.01）.The mRNA expression levels of Vimentin,N-cadherin and Snail were significantly increased in SP cells （9.527-fold,6.834-fold and g.173-fold,respectively） as compared with those in NSP cells （P 〈 0.01）.In contrast,the decrease of E-cadherin mRNA expression in SP cells （5.353-fold） was markedly higher than that in NSP cells （P 〈 0.01）.The protein expression levels of E-cadherin,Vimentin,N-cadherin and Snail in SP cells were markedly different from those in NSP cells （0.174 ±0.014 and 0.935 ±0.012; 1.117 ±0.012 and 0.314 ±0.011; 0.975 ±0.017 and 0.179 ±0.013; 0.917 ±0.014and 0.202 ± 0.013 ; P 〈 0.01）.Conclusion SP cells from hepatoma SMMC-7721 cells have high invasiveness in vitro,which may be related to the changes in the expression of EMT markers.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2014年第1期84-86,共3页
Chinese Journal of Experimental Surgery
基金
河南省科技厅基础与前沿研究项目（082300450100）
关键词
癌
肝细胞
侧群细胞
肿瘤干细胞
侵袭性
上皮-间充质转化
Carcinoma,hepatocellular
Side population cells
Tumor stem cells
Invasion
Epithelial-mesenchymal transition
