中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2012年
9期
1667-1670
,共4页
曾令成%万锋%韩林%叶飞%郭东生%雷霆
曾令成%萬鋒%韓林%葉飛%郭東生%雷霆
증령성%만봉%한림%협비%곽동생%뢰정
CD133%AC133%胶质瘤%脑肿瘤干细胞%分化
CD133%AC133%膠質瘤%腦腫瘤榦細胞%分化
CD133%AC133%효질류%뇌종류간세포%분화
CD133%AC133%Glioma%Brain tumor stem cell%Differentiation
目的 探讨CD133 mRNA及蛋白表达与胶质瘤细胞未分化状态的相关性.方法 对富含AC133表达的胶质瘤干细胞进行了2周诱导分化,实时荧光定量聚合酶链反应(PCR)检测分化前后CD133 mRNA表达,Western blot法检测CD133总蛋白表达,流式细胞学检测细胞表面及细胞内AC133蛋白表达,酶联免疫吸附试验(ELISA)检测培养基上清中AC133蛋白的表达.结果 AC133表达随胶质瘤干细胞分化出现显著下调(P＜0.05),CD133 mRNA定量表达在NCH421k干细胞中分化前为1.08 ±0.39,血清环境下分化为1.08±0.51,血清+1μmol/L全反式维甲酸(ATRA)分化下为1.07 ±0.54,差异无统计学意义(P＞0.05),在NCH441干细胞中分别为2.61±1.72、2.53±1.18、2.18±1.73,差异也无统计学意义(P＞0.05).CD133总蛋白表达在分化前后无显著改变.结论 细胞表面AC133是相对于CD133 mRNA及蛋白,能够更敏感地反映胶质瘤细胞未分化状态的指标.
目的 探討CD133 mRNA及蛋白錶達與膠質瘤細胞未分化狀態的相關性.方法 對富含AC133錶達的膠質瘤榦細胞進行瞭2週誘導分化,實時熒光定量聚閤酶鏈反應(PCR)檢測分化前後CD133 mRNA錶達,Western blot法檢測CD133總蛋白錶達,流式細胞學檢測細胞錶麵及細胞內AC133蛋白錶達,酶聯免疫吸附試驗(ELISA)檢測培養基上清中AC133蛋白的錶達.結果 AC133錶達隨膠質瘤榦細胞分化齣現顯著下調(P＜0.05),CD133 mRNA定量錶達在NCH421k榦細胞中分化前為1.08 ±0.39,血清環境下分化為1.08±0.51,血清+1μmol/L全反式維甲痠(ATRA)分化下為1.07 ±0.54,差異無統計學意義(P＞0.05),在NCH441榦細胞中分彆為2.61±1.72、2.53±1.18、2.18±1.73,差異也無統計學意義(P＞0.05).CD133總蛋白錶達在分化前後無顯著改變.結論 細胞錶麵AC133是相對于CD133 mRNA及蛋白,能夠更敏感地反映膠質瘤細胞未分化狀態的指標.
목적 탐토CD133 mRNA급단백표체여효질류세포미분화상태적상관성.방법 대부함AC133표체적효질류간세포진행료2주유도분화,실시형광정량취합매련반응(PCR)검측분화전후CD133 mRNA표체,Western blot법검측CD133총단백표체,류식세포학검측세포표면급세포내AC133단백표체,매련면역흡부시험(ELISA)검측배양기상청중AC133단백적표체.결과 AC133표체수효질류간세포분화출현현저하조(P＜0.05),CD133 mRNA정량표체재NCH421k간세포중분화전위1.08 ±0.39,혈청배경하분화위1.08±0.51,혈청+1μmol/L전반식유갑산(ATRA)분화하위1.07 ±0.54,차이무통계학의의(P＞0.05),재NCH441간세포중분별위2.61±1.72、2.53±1.18、2.18±1.73,차이야무통계학의의(P＞0.05).CD133총단백표체재분화전후무현저개변.결론 세포표면AC133시상대우CD133 mRNA급단백,능구경민감지반영효질류세포미분화상태적지표.
Objective To explore the correlation between the expression of CD133 and undifferentiated state of glioma cells.Methods Differentiation was induced for 2 weeks in glioma stem cells enriched with AC133 expression.Real-time quantitative polymerase chain reaction (PCR) was performed to study CD133 mRNA expression.Western blotting was used to detect the CD133 protein expression.By using flow cytometry,extracellular and intracellular AC133 expression was examined.Enzyme linked immunosorbent assay (ELISA) was applied to measure the AC133 expression in the culture supernatant.Results The extracellular AC133 expression in glioma stem cells was significantly down-regulated upon differentiation (P＜ 0.05 ). Quantitative CD133 mRNA expression in NCH421k cells before differentiation was 1.08 ±0.39,1.08 ± 0.51 in serum upon differentiation,and 1.07 ± 0.54 in serum plus 1 μmol/L alltrans retinoic acid (P ＞0.05),and that in NCH441 cells was 2.61 ± 1.72,2.53 ± 1.18 and 2.18 ± 1.73respectively (P ＞ 0.05). No significant difference in the CD133 protein expression was observed before and after differentiation.Conclusion Extracelluar AC133 more sensitively mirrors undifferentiated state of glioma cells than CD133 mRNA and protein.
