中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2012年
4期
264-268
,共5页
胃肿瘤%侧群细胞%肿瘤干细胞%流式细胞术%BGC-823细胞%小鼠
胃腫瘤%側群細胞%腫瘤榦細胞%流式細胞術%BGC-823細胞%小鼠
위종류%측군세포%종류간세포%류식세포술%BGC-823세포%소서
Stomach neoplasms%Side population cells%Tumor stem cells%Flow cytometry%BGC-823 cells%Mice
目的 检测和分选胃癌细胞系中具有干细胞特性的侧群(SP)细胞,分析其生物学特性.方法 制备胃癌细胞株BGC-823悬液,经Hoechst 33342染色后,用荧光激活细胞分选技术分选出人胃癌SP和非SP(NSP)细胞,测定SP细胞亚群所占的比例.通过平板克隆实验检测SP细胞的克隆形成能力,无血清培养检测SP细胞的增殖能力.以不同浓度的氟尿嘧啶(5-Fu)处理BGC-823细胞后,检测SP细胞的耐药性,通过小鼠体内成瘤实验检测SP细胞的致瘤性.结果 流式细胞仪分析结果表明,BGC-823细胞中含少量SP细胞,占0.9％ ～2.1％.SP细胞组克隆形成数量为(217.67±15.50)个/孔,高于NSP细胞组[(147.00 ±17.58)个/孔,P＜0.01];SP细胞组克隆形成率为(72.56±5.17)％,高于NSP细胞组[(49.00±5.86)％,P＜0.01],表明SP细胞较NSP细胞有较强的克隆形成能力.经2.5、5、10和25 μg/ml的5-Fu作用36 h后,SP细胞所占比例分别为12.9％、9.9％、4.0％和2.1％,化疗药物对SP细胞的抑制作用随着药物浓度的递增而加强.体内成瘤实验显示,SP细胞组瘤体质量为(0.176 ±0.034)g,NSP细胞组瘤体质量仅为(0.045 ±0.046)g,两组细胞所致瘤体质量差异有统计学意义(P＜0.05).结论 人胃癌细胞BGC-823中存在具有干细胞特性的SP细胞,进一步开展对人胃癌干细胞生物学特性的研究,将可能提示胃癌新的诊断和治疗途径.
目的 檢測和分選胃癌細胞繫中具有榦細胞特性的側群(SP)細胞,分析其生物學特性.方法 製備胃癌細胞株BGC-823懸液,經Hoechst 33342染色後,用熒光激活細胞分選技術分選齣人胃癌SP和非SP(NSP)細胞,測定SP細胞亞群所佔的比例.通過平闆剋隆實驗檢測SP細胞的剋隆形成能力,無血清培養檢測SP細胞的增殖能力.以不同濃度的氟尿嘧啶(5-Fu)處理BGC-823細胞後,檢測SP細胞的耐藥性,通過小鼠體內成瘤實驗檢測SP細胞的緻瘤性.結果 流式細胞儀分析結果錶明,BGC-823細胞中含少量SP細胞,佔0.9％ ～2.1％.SP細胞組剋隆形成數量為(217.67±15.50)箇/孔,高于NSP細胞組[(147.00 ±17.58)箇/孔,P＜0.01];SP細胞組剋隆形成率為(72.56±5.17)％,高于NSP細胞組[(49.00±5.86)％,P＜0.01],錶明SP細胞較NSP細胞有較彊的剋隆形成能力.經2.5、5、10和25 μg/ml的5-Fu作用36 h後,SP細胞所佔比例分彆為12.9％、9.9％、4.0％和2.1％,化療藥物對SP細胞的抑製作用隨著藥物濃度的遞增而加彊.體內成瘤實驗顯示,SP細胞組瘤體質量為(0.176 ±0.034)g,NSP細胞組瘤體質量僅為(0.045 ±0.046)g,兩組細胞所緻瘤體質量差異有統計學意義(P＜0.05).結論 人胃癌細胞BGC-823中存在具有榦細胞特性的SP細胞,進一步開展對人胃癌榦細胞生物學特性的研究,將可能提示胃癌新的診斷和治療途徑.
목적 검측화분선위암세포계중구유간세포특성적측군(SP)세포,분석기생물학특성.방법 제비위암세포주BGC-823현액,경Hoechst 33342염색후,용형광격활세포분선기술분선출인위암SP화비SP(NSP)세포,측정SP세포아군소점적비례.통과평판극륭실험검측SP세포적극륭형성능력,무혈청배양검측SP세포적증식능력.이불동농도적불뇨밀정(5-Fu)처리BGC-823세포후,검측SP세포적내약성,통과소서체내성류실험검측SP세포적치류성.결과 류식세포의분석결과표명,BGC-823세포중함소량SP세포,점0.9％ ～2.1％.SP세포조극륭형성수량위(217.67±15.50)개/공,고우NSP세포조[(147.00 ±17.58)개/공,P＜0.01];SP세포조극륭형성솔위(72.56±5.17)％,고우NSP세포조[(49.00±5.86)％,P＜0.01],표명SP세포교NSP세포유교강적극륭형성능력.경2.5、5、10화25 μg/ml적5-Fu작용36 h후,SP세포소점비례분별위12.9％、9.9％、4.0％화2.1％,화료약물대SP세포적억제작용수착약물농도적체증이가강.체내성류실험현시,SP세포조류체질량위(0.176 ±0.034)g,NSP세포조류체질량부위(0.045 ±0.046)g,량조세포소치류체질량차이유통계학의의(P＜0.05).결론 인위암세포BGC-823중존재구유간세포특성적SP세포,진일보개전대인위암간세포생물학특성적연구,장가능제시위암신적진단화치료도경.
Objective Isolate and characterize the side population (SP) cells with potency of stem cells from human gastric carcinoma cell line BGC-823.Methods SP and non-SP cells were sorted from BGC-823 cells by fluorescence-activated cell sorting (FACS) using Hoechst33342 staining.The tumorigenic ability of the SP cells was assessed by in vivo transplantation into non-obese diabetic/severe combined immunodeficiency mice.Results SP cells were isolated from BGC-823 cells in a proportion of 0.9％ to 2.1％ with respect to the whole cell population.The colony formation assay showed that the colony formation rate of the SP cells was significantly higher than that of the non-SP cells (72.56％ vs.49.00％,P ＜ 0.01 ).The drug sensitivity test showed that the SP cells showed stronger drug resistance to 5-Fu than the non-SP cells.The in vivo transplantation of SP cells in mice showed that the tumor weight was (0.176 ± 0.034) g,significantly higher than that of non-SP cells (0.045 ± 0.046) g ( P ＜ 0.01 ).Conclusions The results of this study indicate the existence of cancer stem-like SP cells in the human gastric cancer line BGC-823 cells. Further characterization of this SP cell population may provide new insights for diagnosis and treatment of gastric cancer.
