Spheroid body-forming cells in the human gastric cancer cell line MKN-45 possess cancer stem cell properties.
Spheroid body-forming cells in the human gastric cancer cell line MKN-45 possess cancer stem cell properties.
复制标题
DOI：
10.3892/ijo.2012.1720
复制
发表时间：
2013-02
影响因子：
5.2
通讯作者：
Zhou Y
中科院分区：
文献类型：
--
作者：
Liu J;Ma L;Xu J;Liu C;Zhang J;Liu J;Chen R;Zhou Y
肿瘤干细胞理论认为，具有自我更新和其他干细胞特性的肿瘤干细胞(CSCs)被认为是肿瘤形成、复发和转移的原因。CSCs的分离和鉴定有助于开发针对CSCs的新的治疗策略。在本研究中，我们通过在特定的无血清培养基中培养人胃癌细胞系MKN-45，通过球形小体的形成来丰富胃癌干细胞。检测MKN-45球体形成细胞的自我更新、增殖、抗药性、致瘤性等干细胞特性，并检测MKN-45球体形成细胞中的干性基因及相关蛋白的表达水平。此外，对球体形成细胞上的干细胞标记进行了免疫荧光染色，以评估干性因素(Oct4、Sox2、Nanog)与建议的CSC标记CD44之间的关系。结果表明，在干细胞条件培养液中培养的胃癌细胞系MKN-45的非贴壁类球体形成细胞与亲代细胞相比具有持续自我更新、广泛增殖、耐药、高致瘤性和高表达CSC相关基因和蛋白(Oct4、Sox2、Nanog和CD44)等胃CSC特性。更重要的是，共表达多能基因Oct4、Sox2和Nanog的CD44阳性细胞可能代表胃肿瘤干细胞。进一步的实验使用更精细的选择标准，如两个或多个标记的组合，将有助于专门鉴定和纯化CSCs。
The cancer stem cell theory hypothesizes that cancer stem cells (CSCs), which possess self-renewal and other stem cell properties, are regarded as the cause of tumor formation, recurrence and metastasis. The isolation and identification of CSCs could help to develop novel therapeutic strategies specifically targeting CSCs. In this study, we enriched gastric cancer stem cells through spheroid body formation by cultivating the human gastric cancer cell line MKN-45 in defined serum-free medium. The stemness characteristics of spheroid body-forming cells, including self-renewal, proliferation, chemoresistance, tumorigenicity of the MKN-45 spheroid body-forming cells were evaluated, and the expression levels of stemness genes and related proteins in the MKN-45 spheroid body-forming cells were assessed. Furthermore, immunofluorescence staining for the stem cell markers on spheroid body-forming cells was examined to evaluate the association between stemness factors (Oct4, Sox2, Nanog) and the proposed CSC marker CD44. Our data demonstrated that non-adherent spheroid body-forming cells from the gastric cancer cell line MKN-45 cultured in stem cell-conditioned medium possessed gastric CSC properties, such as persistent self-renewal, extensive proliferation, drug resistance, high tumorigenic capacity and overexpression of CSC-related genes and proteins (Oct4, Sox2, Nanog and CD44), compared with the parental cells. More importantly, CD44-positive cells co-expressing the pluripotency genes Oct4, Sox2 and Nanog may represent gastric CSCs. Further experiments using more refined selection criteria such as a combination of two or multiple markers would be useful to specifically identify and purify CSCs.
登录
查看更多内容
影响因子：
7.3
作者：
Burkert, J.;Otto, W. R.;Wright, N. A.
通讯作者：
Wright, N. A.
来自原发性人类肿瘤的卵巢癌起始细胞的鉴定和表征
影响因子：
11.2
作者：
Zhang, Shu;Balch, Curt;Nephew, Kenneth P.
通讯作者：
Nephew, Kenneth P.
影响因子：
5.2
作者：
Fujii, Hiromasa;Honoki, Kanya;Takakura, Yoshinori
通讯作者：
Takakura, Yoshinori
影响因子：
4.3
作者：
Brenner, Baruch;Hoshen, Moshe B.;Kushnir, Michal
通讯作者：
Kushnir, Michal
影响因子：
11.2
作者：
Collins, AT;Berry, PA;Maitland, NJ
通讯作者：
Maitland, NJ