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Fig. 1 | Genes & Nutrition

Fig. 1

From: Metformin and soybean-derived bioactive molecules attenuate the expansion of stem cell-like epithelial subpopulation and confer apoptotic sensitivity in human colon cancer cells

Fig. 1

Comparison of human colon cancer cell lines HCT29 and HCT116 for content of cancer stem-like cells. a Single-cell suspensions of HT29 and HCT116 cells were subjected to flow cytometry to assess the relative expression of CD133+CD44+ colon cancer stem-like cells. Representative FACS plots of cells from HT29 (left) or HCT116 (right). b HCT116 cells exhibited a greater percentage of the CD133+CD44+-enriched subpopulation of stem-like subpopulation compared to HT29 cells. Data are mean ± SEM; *P < 0.05 for HCT116 compared to HT29. c CD133+CD44+-stained cells were sorted, collected, and plated for colonosphere formation. Representative pictures of primary colonospheres formed from sorted HT29 (left) and HCT116 (right) cells grown in suspension culture for 5 days. Magnification = 40×, scale bar 50 µm. d CD133+CD44+ epithelial subpopulation (from FACS of HCT116 cells) had greater colonosphere formation compared to that from HT29 cells. Passage 1 (P1) spheres were dissociated to single cells and grown in suspension culture to form secondary spheres (Passage 2; P2). Counting of colonospheres was done at day 5 (for P1) and day 7 (for P2). Data (mean ± SEM) represent the percent of colonospheres formed from 2.5 × 103 plated cells (P1) and from 1 × 103 plated cells (P2), respectively, from two independent experiments, with each experiment carried out in quadruplicates. e Transcript levels of stem cell markers (CD166, CD44), and key genes involved in the maintenance of cancer stem/progenitor cells (NOTCH 1, FASN, PTEN) were quantified by QPCR in FACS-sorted CD133+CD44+–HCT116 and CD133+CD44+–HT29 cells. Transcripts were normalized to a factor derived from the geometric mean of expression values for β-actin ACTB) and cyclophilin A (CYPA). Data (mean ± SEM; n = 5 samples/cell type) were obtained from two independent experiments. *P < 0.05 for HCT116 compared to HT29

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