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Fig. 6 | Cell & Bioscience

Fig. 6

From: SOX4 maintains the stemness of cancer cells via transcriptionally enhancing HDAC1 revealed by comparative proteomics study

Fig. 6

HDAC1 hallmarks colorectal cancer stem cells. a HDAC1 is upregulated in colorectal cancer spheres. The HCT-116 and HT-29 and the primary human colorectal cancer cells were adherent cultured in 6-well plate or suspension cultured in an ultra-low 6-well plate. The spheres from suspension cultured plate were then re-adherent cultured in 6-well plate. The total RNA from these cells was isolated and subjected to qRT-PCR analysis. b HDAC1 is upregulated in CD44 + colorectal cancer cells. CD44 + and CD44- cells were isolated from HCT-116 and HT-29 and primary human colorectal cancer cells by magnetic-activated cell sorting (MACS). The mRNA levels of HDAC1 in CD44 + or CD44- cells were determined by qRT-PCR. c HDAC1 promotes the self-renewal capacity of colorectal cancer cells on serial passage. The HCT-116 and HT-29 cells with or without HDAC1 overexpression were suspension cultured in ultra-low 6 well plates and the sphere number of indicated primary, secondary and tertiary passaged cells was counted after 10 days. d HDAC1 promotes the sphere-forming cell frequency of colorectal cancer cells. HCT-116 and HT-29 cells transfected with lentivirus containing HDAC1-overexpressing plasmid or empty control plasmids were seeded into 96-well U-bottomed culture plates at a density of 10, 5 or 1 cells per well and cultured for 10 days. The sphere-forming cell frequency was calculated by ELDA software. e HDAC1 promotes tumor-initiating cell frequency of colorectal cancer cells. HCT-116 and HT-29 cells transfected with lentivirus containing HDAC1-overexpressing plasmid or empty control plasmids were injected into the subcutaneous tissues of 6-week-old nude mice at a density of 21, 7, 3 cells per mouse. The number of mice developed tumors was counted after 35 days. The tumor-initiating cell frequency was calculated by ELDA software. f HDAC1 promotes the expression of cancer stem cell markers of colorectal cancer cells. The surface protein levels of CD44 and CD133 in HDAC1-overexpressing and control HCT-116 cells were analyzed by flow cytometry. Data are represented as mean ± s.d.; *P<0.05, **P<0.01, ***P<0.001; two-tailed Student’s t-test

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