Isolation And Characterization Of Side Population From Human Breast Cancer Stem Cell

Objective To identify whether the percentages of SP cells have differences in breast cancer tissues, tumor - adjacent tissues, normal tissues and MCF-7 cancer cell line, and investigate the markers between side population cells and non-side population cells from MCF-7 cancer cell line. To assess the relationship between the percentage of SP cells and clinic factors. We hope to future study the breast cancer stem cells.Methods After staining with Hoechst33342 dye and Verapamil, the percentages of SP cells were detected in cell suspension from breast cancer tissues, tumor - adjacent tissues, normal tissues and MCF-7 cancer cell line respectively with the FACS method.After staining with Hoechst33342 dye, side population cells and non-side population cells were sorted by FACS from MCF-7, and these cell markers, antigens and cycles were measured.To analyze the effect of the percentage of SP cells on the prognosis.Results 1. The percentages of SP cells in breast cancer tissues, tumor - adjacent tissues, normal tissues and MCF-7 cancer cell line were (0.54±0.17) %, (7.23±1.12) %, (8.34±0.96) % and (9.58±1.27) %,respectively. There are significant differences between them. 2. There was no marked difference in the proportions of CD38~+, CD117~+,CD34~+ between side population cells and non-side population cells (P=0.256,P=0.920,P=0.317) . The level of CD133~+ and ABCG2~+ in side population cells were higher than the proportions of non-side population cells (P=0.000, P= 0.000) . But the level of CD24~+ was significant lower in side population cells than the proportions of non-side population cells (P=0.000) .3. In side population cells, the proportion of G0/G1 was obviously higherthan that in non-side population cells (P=0.000) . And the proportions of S and G2/M significantly increased in non-side population cells ( P= 0.001, P= 0.000). Compared with non-side population cells, the level of Ki-67 was lower in side population cells. There was significant difference (P=0.000).4. For breast cancer patients, there was a obvious correlation between the SP cells expression and dangerous factors.Conclusion 1. Side population cells express in breast cancer tissues,tumoradjacent tissues,normal tissues and MCF-7 cancer cell line, and the percentages of SP cells are different. This result can supply the scientific value to identify and purify breast cancer stem cells.2. Side population cells express high lever in breast cancer cell line, and SP and non-SP cells can be identified and purified by using their antigenic markers properties. To combine using the SP and antigenic markers properties may be a scientific method to isolate breast cancer stem cells.3. For breast cancer patients, the more high of SP cells expression,the more advenced.