胡 均, 柳建中, 岳 欣, 李 健①, 秦仁义②. 人胆囊癌SP细胞的分离及其干细胞标记物的表达*[J]. 中国肿瘤临床, 2010, 37(1): 44-47. DOI: 10.3969/j.issn.1000-8179.2010.01.012
引用本文: 胡 均, 柳建中, 岳 欣, 李 健①, 秦仁义②. 人胆囊癌SP细胞的分离及其干细胞标记物的表达*[J]. 中国肿瘤临床, 2010, 37(1): 44-47. DOI: 10.3969/j.issn.1000-8179.2010.01.012
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HU Jun1, LIU Jianzhong1, YUE Xin1, LI Jian2, QIN Renyi3. Expression of Stem Cell Marker in Side Population Cells Isolated from Human Gallbladder Carcinoma Cell Line GBC-SD[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2010, 37(1): 44-47. DOI: 10.3969/j.issn.1000-8179.2010.01.012
Citation: HU Jun1, LIU Jianzhong1, YUE Xin1, LI Jian2, QIN Renyi3. Expression of Stem Cell Marker in Side Population Cells Isolated from Human Gallbladder Carcinoma Cell Line GBC-SD[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2010, 37(1): 44-47. DOI: 10.3969/j.issn.1000-8179.2010.01.012
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人胆囊癌SP细胞的分离及其干细胞标记物的表达*

Expression of Stem Cell Marker in Side Population Cells Isolated from Human Gallbladder Carcinoma Cell Line GBC-SD

    摘要
  • 摘要: 目的：探索在人胆囊癌GBC-SD 细胞系中是否存在具有干细胞特性的SP细胞（side population cells），以及SP细胞、非SP细胞和GBC-SD 细胞系表达常见干细胞标记物ABCG2、Oct- 4、CD34的情况。方法：采用荧光激活细胞分选技术（FACS）分选出人胆囊癌SP细胞和非SP细胞，通过RT-PCR、Western blot、流式细胞术以及免疫荧光化学技术检测SP细胞、非SP细胞以及GBC-SD 细胞系表达ABCG2、Oct- 4 和CD34的情况。结果：人胆囊癌GBC-SD 细胞系中存在着少量的具有干细胞潜能的SP细胞，其所占的比例为（0.64± 0.08）% ，SP细胞相对于非SP细胞和GBC-SD 细胞，具有更强的体外侵袭力（P<0.05）；并且ABCG2 在胆囊癌SP细胞中呈现出高表达的状态（89.56±3.86）% ，在非SP细胞中几乎不表达（1.32± 0.49）% ，在未分选胆囊癌细胞系中ABCG2 呈弱表达（12.37± 1.61）% ，P=0.001；Oct- 4 在三种细胞中的表达分别为：（94.87± 1.40）% 、（88.16± 2.34）% 、（90.17± 1.61）% ，P>0.05；CD34在mRNA水平高表达于非SP细胞和GBC-SD 细胞，不表达于SP细胞；在蛋白水平几乎均不表达于这三种细胞，其含量依次为：（1.78± 0.51）% 、（0.63± 0.21）% 、（0.96± 0.38）% ，P>0.05。结论：人胆囊癌GBC-SD 细胞系中存在具有干细胞特性的SP细胞，并且具有ABCG2+Oct- 4+CD34-的表型特征。

     

    Abstract: Objective: To investigate the existence of side population cells with the potency of stem cells in human gallbladder carcinoma cell line GBC-SD and the differences in ABCG 2, Oct- 4 and CD34expression among SP cells, non-SP cells and GBC-SD cells. Methods:SP and non-SP cells were sorted from GBC-SD cells by fluorescence-activated cell sorting (FACS). The expression of ABCG 2, Oct- 4 and CD34in SP cells, non-SP cells, and GBC-SD cells was detected by reverse transcription-polymerase chain reaction (RT-PCR), Western blot, flow cytometry (FCM) and immunofluorescence chemistry. Results: SP cells with stem cell poten -cy were isolated from GBC-SD cells with a proportion of ( 0.64± 0.08)%. The metastatic ability of SP cells was obviously higher than that of non-SP cells and GBC-SD cells ( P<0.05). The expression of ABCG2 was signifi-cantly higher in SP cells than in non-SP cells and GBC-SD cells (89.56± 3.86)% vs. ( 1.32± 0.49)% and ( 12.37±1.61)% , P=0.001 . The expression of Oct- 4 in these cells was ( 94.87± 1.40)% , ( 88.16± 2.34)% and (90.17±1.61)% , respectively (P>0.05). CD 34 was nearly absent in these cells on protein level (1.78± 0.51)% vs. (0.63± 0.21)% and ( 0.96± 0.381 )% , P>0.05), but it was highly expressed in non-SP cells and GBC-SD cells and absent in SP cells on mRNA level.Conclusion:SP cells, which have the potency of stem cells, exist in human gallbladder carcinoma GBC-SD cell line and have the phenotype of ABCG 2+Oct-4+CD34-.

     

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