Characterization of cancer stem-like cells in the side population cells of human gastric cancer cell line MKN-45

人胃癌细胞系MKN-45侧群细胞中癌症干细胞样细胞的表征

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作者:Hai-hong Zhang, Ai-zhen Cai, Xue-ming Wei, Li Ding, Feng-zhi Li, Ai-ming Zheng, Da-jiang Dai, Rong-rong Huang, Hou-jun Cao, Hai-yang Zhou, Jian-mei Wang, Xue-jing Wang, Wei Shi, Heng Zhu, Xiao-ying Yuan, Lin Chen

Conclusions

These results indicate that SP cells possess cancer stem cell properties and prove that SP cells from MKN-45 are gastric cancer stem-like cells.

Methods

We used fluorescence activated cell sorting (FACS) to sort SP cells in the human gastric carcinoma cell line MKN-45 (cells labeled with Hoechst 33342) and then characterized the cancer stem-like properties of SP cells.

Objective

Side population (SP) cells may play a crucial role in tumorigenesis and the recurrence of cancer. Many kinds of cell lines and tissues have demonstrated the presence of SP cells, including several gastric cancer cell lines. This study is aimed to identify the cancer stem-like cells in the SP of gastric cancer cell line MKN-45.

Results

This study found that the SP cells had higher clone formation efficiency than major population (MP) cells. Five stemness-related gene expression profiles, including OCT-4, SOX-2, NANOG, CD44, and adenosine triphosphate (ATP)-binding cassette transporters gene ABCG2, were tested in SP and MP cells using quantitative real-time reverse transcription polymerase chain reaction (RT-PCR). Western blot was used to show the difference of protein expression between SP and MP cells. Both results show that there was significantly higher protein expression in SP cells than in MP cells. When inoculated into non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice, SP cells show higher tumorigenesis tendency than MP cells. Conclusions: These results indicate that SP cells possess cancer stem cell properties and prove that SP cells from MKN-45 are gastric cancer stem-like cells.

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