摘要
目的分析人胃癌细胞系HGC-27细胞中肿瘤干细胞相关的SP（sidepopulation）亚群比例，分选出相关的sP和NSP细胞，并鉴别相关的sP是否具有干细胞某些生物学特性。方法制备人胃癌细胞系HGC-27细胞中细胞悬液，经Hoechst33342和PI染色（其中对照组同时加入拮抗剂维拉帕米），流式细胞仪分析相关SP亚群。通过流式细胞仪分选出人胃癌细胞系HGC-27细胞中相关的sP细胞。克隆形成实验和MTY实验检测相关的sP和NSP细胞增殖能力差异。结果SP亚群占HGC-27细胞中的6．00％左右，维拉帕米阻断后减少为0．01％左右。流式细胞仪分选出sP细胞8．5×105个，大部分为NSP细胞。克隆形成实验和MTY实验证实相关的SP细胞和NSP细胞增殖能力有差异。结论人胃癌细胞系HGC-27细胞中存在相关的sP亚群细胞，维拉帕米可抑制染料外排而明显减少sP细胞的比例。可以通过流式细胞仪分选出相关的sP细胞，通过克隆形成实验和MTr实验检测相关的sP和NSP细胞增殖能力有差异。但尚需从分子水平证实胃癌细胞SP细胞相关干细胞特异性。
Objective To identify the ratio of the side population （SP） of cancer stem-like cells in gastric cancer cell line I-IGC-27 and separate the SP and NSP cells. To initially identify whether the SP cells have some stem- cell bionomics. Methods HGC-27 cells were stained with Hoechst33342 and PI in the absence or presence of vera- pamil. Then, the SP was separated and analyzed by flow cytometry. The proliferation of SP and NSP cells was detected by colony formation assay and M33＂ assay. Results SP cells accounted for 6.00% of the HGC-27 cells, which de- creased to 0.01% treatment with verapamil. 8.5 x 105 SP cells were separated by the flow cytometry. The results of colony formation assay and MTT assay showed that the proliferation of SP and NSP cells was significantly different. Conclusion Human gastric cancer cell line HGC-27 contains SP,which can be decreased by verapamil. SP cells can be separated by flow cytometry, The colony formation assay and MTT assay confirmed the difference of SP and NSP cells in proliferation. But we should identify the differences between the SP and NSP cells from molecular level.
出处
《中国基层医药》
CAS
2013年第24期3681-3683,共3页
Chinese Journal of Primary Medicine and Pharmacy
基金
上海市卫生局科学基金资助项目（2010Y079）
