空军医学杂志
空軍醫學雜誌
공군의학잡지
MEDICAL JOURNAL OF AIR FORCE
2013年
1期
36-40
,共5页
张海宏%魏学明%范勤%王建梅%王雪晶%李丽%石玮%袁小英%陈凛
張海宏%魏學明%範勤%王建梅%王雪晶%李麗%石瑋%袁小英%陳凜
장해굉%위학명%범근%왕건매%왕설정%리려%석위%원소영%진름
ATP结合转运盒%侧群细胞%苯咪唑类 (Hoe 33342)%胃癌%干细胞
ATP結閤轉運盒%側群細胞%苯咪唑類 (Hoe 33342)%胃癌%榦細胞
ATP결합전운합%측군세포%분미서류 (Hoe 33342)%위암%간세포
ATP-Binding cassette transporters%Side-population cells%Benzimidazoles (hoe 33342)%Stomach neoplasm%Stem cells
目的探讨胃癌细胞系中是否存在SP细胞及SP细胞的生物学特征。方法采用荧光活化的细胞分选技术(FACS)从胃癌MKN-45细胞株获得SP细胞,进而分析这些SP细胞有无肿瘤干细胞特性;采用实时定量的RT-PCR检测5个干性相关基因OCT-4, SOX-2, NANOG, CD44和ABCG-2的mRNA在SP和MP 细胞中的表达水平,并进行统计学差异分析。结果发现二者之间具有显著差异。进一步使用蛋白印迹法(Western blot)定性分析发现:5个干性相关基因在SP和MP细胞中的蛋白表达水平亦有差异;体内实验结果显示,当SP细胞和MP细胞注射非糖尿病联合重度免疫缺陷(NOD/SCID)的小鼠,SP 细胞较MP细胞具有更强的成瘤性。结论综合分析本实验相关结果,可以推测SP细胞具有肿瘤干细胞特性,由MKN-45分选获得的SP细胞是具有干细胞特性的胃癌干样细胞。SP细胞较主群细胞(MP)具有较高的克隆形成率。
目的探討胃癌細胞繫中是否存在SP細胞及SP細胞的生物學特徵。方法採用熒光活化的細胞分選技術(FACS)從胃癌MKN-45細胞株穫得SP細胞,進而分析這些SP細胞有無腫瘤榦細胞特性;採用實時定量的RT-PCR檢測5箇榦性相關基因OCT-4, SOX-2, NANOG, CD44和ABCG-2的mRNA在SP和MP 細胞中的錶達水平,併進行統計學差異分析。結果髮現二者之間具有顯著差異。進一步使用蛋白印跡法(Western blot)定性分析髮現:5箇榦性相關基因在SP和MP細胞中的蛋白錶達水平亦有差異;體內實驗結果顯示,噹SP細胞和MP細胞註射非糖尿病聯閤重度免疫缺陷(NOD/SCID)的小鼠,SP 細胞較MP細胞具有更彊的成瘤性。結論綜閤分析本實驗相關結果,可以推測SP細胞具有腫瘤榦細胞特性,由MKN-45分選穫得的SP細胞是具有榦細胞特性的胃癌榦樣細胞。SP細胞較主群細胞(MP)具有較高的剋隆形成率。
목적탐토위암세포계중시부존재SP세포급SP세포적생물학특정。방법채용형광활화적세포분선기술(FACS)종위암MKN-45세포주획득SP세포,진이분석저사SP세포유무종류간세포특성;채용실시정량적RT-PCR검측5개간성상관기인OCT-4, SOX-2, NANOG, CD44화ABCG-2적mRNA재SP화MP 세포중적표체수평,병진행통계학차이분석。결과발현이자지간구유현저차이。진일보사용단백인적법(Western blot)정성분석발현:5개간성상관기인재SP화MP세포중적단백표체수평역유차이;체내실험결과현시,당SP세포화MP세포주사비당뇨병연합중도면역결함(NOD/SCID)적소서,SP 세포교MP세포구유경강적성류성。결론종합분석본실험상관결과,가이추측SP세포구유종류간세포특성,유MKN-45분선획득적SP세포시구유간세포특성적위암간양세포。SP세포교주군세포(MP)구유교고적극륭형성솔。
Objective We tried to ascertain gastric cancer cell line MKN-45 contain SP cells, and learned about wether gastric cancer SP cells possess cancer stem-like characteristic. Method We used fluorescence activated cell sorting (FACS) to sort SP cells in human gastric carcinoma (GC) cell line MKN-45 cells labeled with Hoechst 33342, and then characterized the cancer stem-like properties of SP cells. Result The SP cells had higher clone formation efficiency than major population (MP) cells. Five stemness–related genes expression profiles, including OCT-4, SOX-2, NANOG, CD44 and ATP-binding cassette transporters gene ABCG-2, were tested in SP and MP cells using quantitative real-time RT-PCR. Western blot was chosen to show the difference of protein expression between SP and MP cells. Both results showed that there was significantly higher expression in SP cells than in MP cells. When inoculated into non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice, SP cells showed higher tumorigenesis tendency than MP cells. Conclusion SP cells possess cancer stem cell properties and proved that SP cells from MKN-45 were gastric cancer stem-like cells.