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Enriching effect of 5-fluorouracil on cancer stem cell-like cells from mouse breast cancer cell line 4T1
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Abstract:
Objective:To explore the possibility of enriching breast cancer stem cell-like cells by cell passage in a tumor-bearing mouse model treated with 5-fluorouracil （5-FU） in vivo and to lay a foundation for cancer stem cell-targeted therapy. Methods: To establish a tumor-bearing model by inoculating breast cancer cell line 4T1 into mouse subcutaneously. 5-FU was injected into these mice intraperitonealy according to different groups. The mice were executed 4 weeks after injection. Cell suspension made from mouse tumor was injected into the mice again to establish the next generation of tumor-bearing mouse model. The mice were treated with 5-FU in the third and fourth generation as above. Four generations of the mouse model were established in all. The mice in control group were injected with normal saline instead of 5-FU, and the following four generation tumor-bearing models were treated as 5-FU group. Flow cytometry was used to measure the proportion of CD44+CD24-/low cells in different tumor tissues; Hoechast 33342 staining was used to measure the proportion of SP (side population) cells; and expressions of ALDH1 and CD55 proteins were detected by immunohistochemistry. The formation of mammospheres was observed under an inverted microscope. Tumorigenic ability of different tumor cells was detected by mouse tumorigenesis experiment. Results: In the control group, the proportion of CD44+CD24-/lowcells in mouse tumor was (11.5±0.9)%, and the proportion of SP cells was (9.7±13)%. The expression level of ALDH1 was negative, the proportion of cells in which CD55 was highly expressed, was (0.6±0.3)%, and the proportion of mammospheres was (0.5±0.2)%. While in 5-FU treated group, the proportions of CD44+CD24-/lowcells were respectively (49.8±1.2)%, (56.8±1.7)%, (66.4±1.5)% and (69.0±1.6)%; the proportions of SP cells were respectively (25.0±1.2)%, (42.6±2.8)%, (58.4±2.1)% and (61.3±2.6)%; the proportions of cells in which ALDH1 was positively expressed, were respectively (3.8±0.7)%, (14.1±24)%, (25.2±31)% and (275±2.7)%, and the proportions of cells in which CD55 were highly expressed, were respectively (7.8±16)%, (10.1±2.0)%, (15.6±1.4)% and (17.3±1.9)%, and the proportion of mammospheres was (5.9±04)%. The significant differences were found between the two groups（P<0.05 or P<0.01）. The third generation tumor cells enriching cancer stem cells were more tumorigenic than the control tumor cells. Conclusion: 5-FU enrichs the cancer stem cell-like cells in mouse breast cancer cell line 4T1 by the cell passage in the tumor-bearing mouse model in vivo.
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Project Supported:
Project supported by the Natural Science Foundation of Hebei Province (No. 2012206135), and the Science and Technology Research Program of Xingtai City (No. 2012-ZC-085)
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