Comparison of enrichment model of cancer stem cells in colon cancer DLD-1 cells

• DOI: 10.3760/cma.j.issn.1673-9752.2016.02.011
• Publication Date: 2016-02-20
• Authors: Zhengwei Leng; Xiaorong Zhong; Yongfu Xiong; Yonghong Zhou; Wei Zhong; Zhipeng Zhan; Hongyu Lin
• Keywords: Homeobox protein NANOG

Objective To compare the effects of 3 enrichment models cancer stem cells (CSCs) in colon DLD-1 cells.

Methods DLD-1 were cultured kinds serum-free medium containing various growth factors to generate spheroid cells, including G1 group using 10 ng/mL EGF+ bFGF, G2 20 bFGF and G3 bFGF. The cell proportion CD133+ CD44+ , ALDH+ confirmed by flow cytometery. mRNA expression related genes (Oct4, Sox2, Nanog) CSCs (CD133, CD44, CD166, ALDH, CD66c) detected real-time polymerase chain reaction (RT-PCR). capacity self-renewal was sphere-forming assay. tumorigenesis vivo measured colony formation Measurement data with normal distribution presented as ±s, comparisons among groups analyzed one-way ANOVA, pairwise done paired-samples t test.

Results The results cytometery: G1, 0.030±0.010, 0.113±0.012 0.043±0.006, respectively, a significant difference (F=67.625, P 0.05). respectively 0.037±0.015, 0.027±0.006 0.023±0.006, no (F=1.444, P>0.05). proportions 20.103±4.761, 44.600±2.138 44.387±2.117, (F=56.269, 0.060±0.010, 0.627±0.068 0.043±0.021, (F=192.097, RT-PCR: expressions 1.272±0.152, 11.636±3.164 1.420±0.201 Oct4, 4.717±0.409, 8.262±1.079 4.827±0.446 1.836±0.417, 12.883±4.030 1.636± 0.589 Nanog, 1.555±0.397, 4.367±1.109 1.208±0.165 CD133, 25.853±4.544, 59.483±8.950 26.924±4.134 21.223±1.088, 57.847±5.880 21.710±4.010 19.378±2.479, 78.211±9.879 18.475±3.571 4.737±2.105, 18.003±3.826 5.211±1.474 CD66c, showing differences (F=31.529, 23.865, 21.557, 19.102, 27.837, 76.608, 90.530, 23.997, assay: numbers spheres 182±34, 396±22 217±22, (F=56.128, colonies 230±33, 403±32 266±36, (F=22.111, 0.05).

Conclusion The culturing could effectively promote cells.

Key words: Colonic neoplasms; Neoplastic cells; Enrichment; Suspension culture