中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2011年
1期
65-67
,共3页
黄涛%宫东伟%高全立%张旭华%吕晓东%周进学
黃濤%宮東偉%高全立%張旭華%呂曉東%週進學
황도%궁동위%고전립%장욱화%려효동%주진학
癌,肝细胞%侧群细胞%肿瘤干细胞
癌,肝細胞%側群細胞%腫瘤榦細胞
암,간세포%측군세포%종류간세포
Carcinoma,hepatocellular%Side population cell%Tumor stem cell
目的 分选肝癌细胞株SMMC-7721中的侧群(SP)细胞,并分析其干细胞标记的表达.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,以实时荧光定量聚合酶链反应(real-time PCR)技术和流式细胞术对两个亚群细胞干细胞标记mRNA和蛋白表达进行分析.结果 SMMC-7721细胞株中分选出的SP细胞比例为(9.2±0.2)%.SP细胞ABCG2、CD133、Oct4、Sox2和NANOG等干细胞标记mRNA的表达水平分别是NSP细胞的7.132倍、4.985倍、8.642倍、5.095倍和5.164倍,差异均有统计学意义(P＜0.01);ABCG2、CD133、Oct4、Sox2和NANOG蛋白在肝癌SP细胞中的含量分别为(92.65±3.92)%、(12.75±1.62)%、(17.35±2.31)%、(9.57±1.71)%和(28.39±5.28)%,在NSP细胞中的含量分别为(0.26±0.06)%、(2.51±0.17)%、(1.74±0.38)%、(1.52±0.41)%和(3.37±1.02)%,差异有统计学意义(P＜0.01).结论 肝癌SMMC-7721细胞中的SP细胞可能富集了肝癌干细胞,联合应用多种干细胞标记筛选肝癌SP细胞可能会获得纯化的肝癌干细胞.
目的 分選肝癌細胞株SMMC-7721中的側群(SP)細胞,併分析其榦細胞標記的錶達.方法 採用流式細胞熒光激活分選(FACS)技術將SMMC-7721細胞分為SP細胞和非側群(NSP)細胞兩箇亞群,以實時熒光定量聚閤酶鏈反應(real-time PCR)技術和流式細胞術對兩箇亞群細胞榦細胞標記mRNA和蛋白錶達進行分析.結果 SMMC-7721細胞株中分選齣的SP細胞比例為(9.2±0.2)%.SP細胞ABCG2、CD133、Oct4、Sox2和NANOG等榦細胞標記mRNA的錶達水平分彆是NSP細胞的7.132倍、4.985倍、8.642倍、5.095倍和5.164倍,差異均有統計學意義(P＜0.01);ABCG2、CD133、Oct4、Sox2和NANOG蛋白在肝癌SP細胞中的含量分彆為(92.65±3.92)%、(12.75±1.62)%、(17.35±2.31)%、(9.57±1.71)%和(28.39±5.28)%,在NSP細胞中的含量分彆為(0.26±0.06)%、(2.51±0.17)%、(1.74±0.38)%、(1.52±0.41)%和(3.37±1.02)%,差異有統計學意義(P＜0.01).結論 肝癌SMMC-7721細胞中的SP細胞可能富集瞭肝癌榦細胞,聯閤應用多種榦細胞標記篩選肝癌SP細胞可能會穫得純化的肝癌榦細胞.
목적 분선간암세포주SMMC-7721중적측군(SP)세포,병분석기간세포표기적표체.방법 채용류식세포형광격활분선(FACS)기술장SMMC-7721세포분위SP세포화비측군(NSP)세포량개아군,이실시형광정량취합매련반응(real-time PCR)기술화류식세포술대량개아군세포간세포표기mRNA화단백표체진행분석.결과 SMMC-7721세포주중분선출적SP세포비례위(9.2±0.2)%.SP세포ABCG2、CD133、Oct4、Sox2화NANOG등간세포표기mRNA적표체수평분별시NSP세포적7.132배、4.985배、8.642배、5.095배화5.164배,차이균유통계학의의(P＜0.01);ABCG2、CD133、Oct4、Sox2화NANOG단백재간암SP세포중적함량분별위(92.65±3.92)%、(12.75±1.62)%、(17.35±2.31)%、(9.57±1.71)%화(28.39±5.28)%,재NSP세포중적함량분별위(0.26±0.06)%、(2.51±0.17)%、(1.74±0.38)%、(1.52±0.41)%화(3.37±1.02)%,차이유통계학의의(P＜0.01).결론 간암SMMC-7721세포중적SP세포가능부집료간암간세포,연합응용다충간세포표기사선간암SP세포가능회획득순화적간암간세포.
Objective To study the expression of stem cell markers in side population cells sorted from SMMC-7721 cell line. Methods Fluorescence-activated cell sorting (FACS) was used to sort side population (SP) cells and non-SP (NSP) cells from SMMC-7721 cell line. Real-time polymerase chain reaction (PCR) and flow cytometry (FCM) were used to evaluate the expression of several stem cell markers such as ABCG2, CD133, Oct4, Sox2 and NANOG in SP cells and NSP cells. Results FACS analysis indicated that (9.2 ±0. 2)% of the SMMC-7721 cells were SP cells. Real-time PCR analysis suggested that ABCG2, CD133, Oct4, Sox2 and NANOG were expressed in the SP cells at higher levels than the NSP cells by about 7. 132, 4. 985, 8. 642, 5.095 and 5. 164 folds, respectively ( P ＜0. 01 ). FCM analysis revealed that the expression of ABCG2, CD133, Oct4, Sox2 and NANOG proteins in SP cells was (92. 65 ±3.92)%, (12.75 ±1.62)%, (17.35 ±2.31)%, (9.57 ± 1.71)% and (28.39 ±5.28)% respectively,while in NSP cells that was (0. 26 ±0. 06)%, (2. 51 ±0. 17)%, ( 1.74 ±0. 38)%, ( 1.52 ±0. 41 )% and ( 3.37 ± 1.02) % respectively ( P ＜ 0. 01 ). Conclusion The SP cells sorted from SMMC-7721 cell line may enrich tumor stem cells. Purified liver cancer stem cells may be obtained by screening SP cells using a variety of stem cell markers.
