天津医药

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滨海新区地区宫颈癌肿瘤干细胞样侧群细胞的恶性行为特征分析

李晓文1,袁俐1,张俊平2,李建勇1,姜忠敏1,刘晓智1   

  1. 1. 天津市第五中心医院
    2. 天津市第五中心医院妇产科
  • 收稿日期:2013-03-25 修回日期:2013-10-28 出版日期:2014-03-15 发布日期:2014-03-15
  • 通讯作者: 袁俐

The analysis of malignant characteristics of tumor stem cell-like side population cells from cervical cancer patients of Binhai New Area

  • Received:2013-03-25 Revised:2013-10-28 Published:2014-03-15 Online:2014-03-15

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摘要： 【摘要】 目的 分析来自滨海新区地区宫颈癌病人群的宫颈癌肿瘤干细胞样侧群细胞的恶性行为特点。方法 自术中切除的宫颈癌组织中原代培养宫颈癌细胞，流式细胞术分离宫颈癌干细胞样侧群与非侧群细胞，MTT法绘制细胞生长曲线，Transwell培养体系比较两群细胞的侵袭能力，软琼脂克隆实验检测瘤细胞的克隆能力，免疫荧光方法检测耐药基因ATP结合体超家族成员2（ATP-binding cassette superfamily G member 2，ABCG2）的蛋白表达情况；去胸腺小鼠腹股沟皮下接种两类瘤细胞，比较成瘤率。结果 流式细胞检测显示宫颈癌干细胞样侧群细胞占细胞总数的1.39％；与非侧群细胞比较，侧群细胞生长迅速，具有更强的瘤细胞侵袭力和克隆形成能力；侧群细胞表达更高水平的ABCG2；小鼠体内成瘤率达100%（10/10），而非侧群细胞为20%（2/10）。结论 宫颈癌肿瘤干细胞样侧群细胞具有更高的恶性细胞表型，是未来宫颈癌基因治疗的核心靶点。

关键词: 宫颈癌, 肿瘤干细胞, 侵袭, 耐药性

Abstract: 【Abstract】 Objective To analysis the malignant performance characteristics of tumor stem cell-like side population cells obtained from cervical cancer patients of Binhai New Area. Methods The cervical cancer cells were obtained from surgical resection tumor tissue, then tumor stem cell-like side population cells were isolated by flow cytometry.The cell growth curve was drawn by MTT assay, the invasion ability of tumor cells was compared by transwell assay, then the clonogenic capacity was detected by clone formation in soft agar. The protein level of ABCG2, which is a drug-resistant gene, was detected by immunofluorescence method. Finally, these cells were transplated into the subcutaneous of de-thymus mice and the tumor form rate was surveyed. Results The results from flow cytometry assay showed the percentage of cervical cancer stem cell-like side population cells was 1.39 %. Compared with the non-side population cells, the side population cells grow quickly, have enhanced invasion ability and colony forming ability, and the side population cells have higher expression level of ABCG2.The tumor form rate of the side population cells was 100% (10/10), but the non-side population cells was 20% (2/10). Conclusion The cervical cancer stem cell-like side population cells have more malignant performance characteristics than the non-side population cells, it maybe become a core target for cancer gene therapy in the future.

Key words: Cervical cancer, Cancer stem cells, Invasion, Drug resistance