上海交通大学学报（医学版）

›› 2011, Vol. 31 ›› Issue (8): 1113-.doi: 10.3969/j.issn.1674-8115.2011.08.015

• 论著（基础研究） • 上一篇    下一篇

肠癌SW480肿瘤细胞SP亚群分析

王 兵1, 刘文勇1, 叶冬霞2, 高钢龙1, 程海冰3   

  1. 上海交通大学 医学院附属第九人民医院 1.普外科, 2.口腔颌面外科, 3.临床医学院, 上海 200011
  • 出版日期:2011-08-28 发布日期:2011-08-29
  • 通讯作者: 刘文勇, 电子信箱: liuwy@163.com。
  • 作者简介:王 兵（1968—）, 男, 副主任医师, 博士；电子信箱: wingping01@126.com。
  • 基金资助:

    上海市科委基金(08JC1414200)；上海市卫生局基金 (2007157)；国家级大学生创新性实验基金(081024850)

Analysis of SP subgroup of colon cancer cell line SW480

WANG Bing1, LIU Wen-yong1, YE Dong-xia2, GAO Gang-long1, CHENG Hai-bing3   

  1. 1.Department of General Surgery, 2.Department of Oral and Maxillofacial Surgery, 3.College of Clinical Medicine, the Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, China
  • Online:2011-08-28 Published:2011-08-29
  • Supported by:

    Shanghai Science and Technology Committee Foundation, 08JC1414200；Shanghai Municipal Health Bureau Foundation, 2007157；National College Student Innovation Foundation, 081024850

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摘要：

目的 分选肠癌细胞株SW480中边群细胞（SP）亚群，观察SP细胞是否具有肿瘤干细胞样生物学特性。方法 将1×107个/mL对数期生长的SW480细胞进行Hoechest33342和PI染色，80 μg/mL维拉帕米阻断，流式细胞仪分选SP细胞。将分选后的SP细胞和非SP细胞，采用MTT法绘制细胞生长曲线和进行细胞克隆形成实验；并将两种细胞接种于裸鼠皮下，观察移植瘤的形成情况。结果 Hoechest33342染色显示：SP细胞亚群的平均含量为（1.43±0.05）%，加入维拉帕米后SP细胞亚群比例减少为（0.01±0.01）%，两组比较差异有统计学意义（P<0.05）。细胞生长曲线显示：非SP组细胞生长速度较SP组明显降低，两组比较差异有统计学意义（P<0.05）。克隆形成实验表明，移植第15天后SP细胞克隆体积大，边界平滑，细胞呈巢状隆起生长，结构致密，细胞体积小，平均克隆数为49.33±8.21，显著高于非SP细胞组的31.03±5.29，两组比较差异也有统计学意义（P<0.01）。在移植量为1×105个SP细胞组中有50%小鼠成瘤，而非SP细胞组小鼠均未见明显成瘤（P<0.01）。结论 人肠癌细胞株SW480中存在SP亚群细胞，生长曲线和克隆形成实验表明其增殖能力较非SP细胞强；裸鼠成瘤试验提示SP细胞尚需进一步纯化。

关键词: SW480细胞株, SP亚群, 肿瘤干细胞, 肠癌

Abstract:

Objective To sort the side population (SP) subgroup in colon cancer cell line SW480, and explore whether SP cells have biological properties of tumor stem cells. Methods Hoechst33342 and PI staining was performed on 1×107/mL SW480 cells of logarithmic growth phase, and cells were treated with 80 μg/mL verapamil. SP cells were sorted by flow cytometry. Cell growth curves of SP cells and non-SP cells were drawn with MTT method, and clonogenic assay was performed. These two cells were subcutaneously inoculated into nude mice, and the tumorigenesis was observed. Results It was revealed by Hoechest33342 staining that the percent of SP subgroup was (1.43±0.05)%, and was decreased to (0.01±0.01)% after treatment with verapamil (P<0.05). Cell growth curves indicated that the growth rate in non-SP group was significantly lower than that in SP group (P<0.05). It was demonstrated by clonogenic assay that 15 d after inoculation, the number of cell clones in SP group (49.33±8.21) was significantly larger than that in non-SP group (31.03±5.29)(P<0.01). In 1×105 SP inoculation group, tumorigenesis was observed in 50% of mice, while there was no tumorigenesis in non-SP group (P<0.01). Conclusion There exists SP subgroup in human SW480 cells, whose proliferation ability may be higher than that of non-SP cells. Further experiments should be performed to purify SP cells.

Key words: SW480 cell line, SP subgroup, tumor stem cells, colon cancer