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Cancer-initiating cells derived from human rectal adenocarcinoma tissues carry mesenchymal phenotypes and resist drug therapies.

Fan CW, Chen T, Shang YN, Gu YZ, Zhang SL, Lu R, OuYang SR, Zhou X, Li Y, Meng WT, Hu JK, Lu Y, Sun XF, Bu H, Zhou ZG, Mo XM - Cell Death Dis (2013)

Bottom Line: These R-CICs generated tumors similar to their tumor of origin when injected into immunodeficient mice, differentiated into rectal epithelial cells in vitro, and were capable of self-renewal both in vitro and in vivo.More importantly, subpopulations of R-CICs resisted both 5-fluorouracil/calcium folinate/oxaliplatin (FolFox) and cetuximab treatment, which are the most common therapeutic regimens used for patients with advanced or metastatic rectal cancer.Thus, the identification, expansion, and properties of R-CICs provide an ideal cellular model to further investigate tumor progression and determine therapeutic resistance in these patients.

View Article: PubMed Central - PubMed

Affiliation: 1] Institute of Digestive Surgery, West China Hospital, Sichuan University, Chengdu, People's Republic of China [2] Medical Center of Stem Cell Biology, West China Hospital, Sichuan University, Chengdu, People's Republic of China [3] Department of Gastrointestinal Surgery, West China Hospital, Sichuan University, Chengdu, People's Republic of China.

ABSTRACT
Accumulating evidence indicates that cancer-initiating cells (CICs) are responsible for cancer initiation, relapse, and metastasis. Colorectal carcinoma (CRC) is typically classified into proximal colon, distal colon, and rectal cancer. The gradual changes in CRC molecular features within the bowel may have considerable implications in colon and rectal CICs. Unfortunately, limited information is available on CICs derived from rectal cancer, although colon CICs have been described. Here we identified rectal CICs (R-CICs) that possess differentiation potential in tumors derived from patients with rectal adenocarcinoma. The R-CICs carried both CD44 and CD54 surface markers, while R-CICs and their immediate progenies carried potential epithelial-mesenchymal transition characteristics. These R-CICs generated tumors similar to their tumor of origin when injected into immunodeficient mice, differentiated into rectal epithelial cells in vitro, and were capable of self-renewal both in vitro and in vivo. More importantly, subpopulations of R-CICs resisted both 5-fluorouracil/calcium folinate/oxaliplatin (FolFox) and cetuximab treatment, which are the most common therapeutic regimens used for patients with advanced or metastatic rectal cancer. Thus, the identification, expansion, and properties of R-CICs provide an ideal cellular model to further investigate tumor progression and determine therapeutic resistance in these patients.

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CD44+CD54+ R-CICs are resistant to chemotherapeutic agents and apoptosis in vitro. (a) Examination of k-ras gene mutation with sequencing, where codons 12 and 13 of exon 2 are wild type. One representative graph of three different tumors is shown. (b) Immunoblotting or immunohistochemical validation of EGFR expression in different cell populations derived from two independent experiments (patients 9 and 15). (c) Percentage of cell viability of different cellular subpopulations and caco-2 cells after treatment with 5-Fu, oxaliplatin, and cetuximab of different concentrations for 24, 48, and 72 h in vitro. Data are mean±S.D. of three independent experiments, each performed with cells from different donors (**P<0.01, *P<0.05; patients 9, 11, and 15). (d) Cell apoptosis percentage of different cellular subpopulations and caco-2 cells after treatment with 5-Fu, oxaliplatin, or cetuximab for 24, 48, or 72 h. Data are mean±S.D. of three independent experiments, each performed with cells from different donors (**P<0.01, *P<0.05; patients 9,11, and 15)
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fig5: CD44+CD54+ R-CICs are resistant to chemotherapeutic agents and apoptosis in vitro. (a) Examination of k-ras gene mutation with sequencing, where codons 12 and 13 of exon 2 are wild type. One representative graph of three different tumors is shown. (b) Immunoblotting or immunohistochemical validation of EGFR expression in different cell populations derived from two independent experiments (patients 9 and 15). (c) Percentage of cell viability of different cellular subpopulations and caco-2 cells after treatment with 5-Fu, oxaliplatin, and cetuximab of different concentrations for 24, 48, and 72 h in vitro. Data are mean±S.D. of three independent experiments, each performed with cells from different donors (**P<0.01, *P<0.05; patients 9, 11, and 15). (d) Cell apoptosis percentage of different cellular subpopulations and caco-2 cells after treatment with 5-Fu, oxaliplatin, or cetuximab for 24, 48, or 72 h. Data are mean±S.D. of three independent experiments, each performed with cells from different donors (**P<0.01, *P<0.05; patients 9,11, and 15)

Mentions: As CICs derived from various solid tumors have been shown to be resistant to chemotherapy,6, 7, 20 we assessed the changes in expression of CD44 and CD54 after culturing rectospherical cells in medium with 5-fluorouracil (5-Fu), oxaliplatin, and cetuximab for 7 days. Cetuximab is a monoclonal antibody that targets epidermal growth factor receptor (EGFR) but exhibits better therapeutic efficacy in k-ras wild-type CRC. Therefore, we also examined EGFR expression and the k-ras mutation in tumor samples and different cellular subpopulations. The k-ras mutation was not detected in the samples assessed (Figure 5a), but the expression of EGFR was observed (Figure 5b). Following treatment with 5-Fu, oxaliplatin, and cetuximab, the fraction of CD44+CD54+ cells significantly increased (Supplementary Figures S2a and b) while the spheroids clearly decreased in number (Supplementary Figure S2c) compared with controls, indicating that this fraction may be resistant to these agents.


Cancer-initiating cells derived from human rectal adenocarcinoma tissues carry mesenchymal phenotypes and resist drug therapies.

Fan CW, Chen T, Shang YN, Gu YZ, Zhang SL, Lu R, OuYang SR, Zhou X, Li Y, Meng WT, Hu JK, Lu Y, Sun XF, Bu H, Zhou ZG, Mo XM - Cell Death Dis (2013)

CD44+CD54+ R-CICs are resistant to chemotherapeutic agents and apoptosis in vitro. (a) Examination of k-ras gene mutation with sequencing, where codons 12 and 13 of exon 2 are wild type. One representative graph of three different tumors is shown. (b) Immunoblotting or immunohistochemical validation of EGFR expression in different cell populations derived from two independent experiments (patients 9 and 15). (c) Percentage of cell viability of different cellular subpopulations and caco-2 cells after treatment with 5-Fu, oxaliplatin, and cetuximab of different concentrations for 24, 48, and 72 h in vitro. Data are mean±S.D. of three independent experiments, each performed with cells from different donors (**P<0.01, *P<0.05; patients 9, 11, and 15). (d) Cell apoptosis percentage of different cellular subpopulations and caco-2 cells after treatment with 5-Fu, oxaliplatin, or cetuximab for 24, 48, or 72 h. Data are mean±S.D. of three independent experiments, each performed with cells from different donors (**P<0.01, *P<0.05; patients 9,11, and 15)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3824647&req=5

fig5: CD44+CD54+ R-CICs are resistant to chemotherapeutic agents and apoptosis in vitro. (a) Examination of k-ras gene mutation with sequencing, where codons 12 and 13 of exon 2 are wild type. One representative graph of three different tumors is shown. (b) Immunoblotting or immunohistochemical validation of EGFR expression in different cell populations derived from two independent experiments (patients 9 and 15). (c) Percentage of cell viability of different cellular subpopulations and caco-2 cells after treatment with 5-Fu, oxaliplatin, and cetuximab of different concentrations for 24, 48, and 72 h in vitro. Data are mean±S.D. of three independent experiments, each performed with cells from different donors (**P<0.01, *P<0.05; patients 9, 11, and 15). (d) Cell apoptosis percentage of different cellular subpopulations and caco-2 cells after treatment with 5-Fu, oxaliplatin, or cetuximab for 24, 48, or 72 h. Data are mean±S.D. of three independent experiments, each performed with cells from different donors (**P<0.01, *P<0.05; patients 9,11, and 15)
Mentions: As CICs derived from various solid tumors have been shown to be resistant to chemotherapy,6, 7, 20 we assessed the changes in expression of CD44 and CD54 after culturing rectospherical cells in medium with 5-fluorouracil (5-Fu), oxaliplatin, and cetuximab for 7 days. Cetuximab is a monoclonal antibody that targets epidermal growth factor receptor (EGFR) but exhibits better therapeutic efficacy in k-ras wild-type CRC. Therefore, we also examined EGFR expression and the k-ras mutation in tumor samples and different cellular subpopulations. The k-ras mutation was not detected in the samples assessed (Figure 5a), but the expression of EGFR was observed (Figure 5b). Following treatment with 5-Fu, oxaliplatin, and cetuximab, the fraction of CD44+CD54+ cells significantly increased (Supplementary Figures S2a and b) while the spheroids clearly decreased in number (Supplementary Figure S2c) compared with controls, indicating that this fraction may be resistant to these agents.

Bottom Line: These R-CICs generated tumors similar to their tumor of origin when injected into immunodeficient mice, differentiated into rectal epithelial cells in vitro, and were capable of self-renewal both in vitro and in vivo.More importantly, subpopulations of R-CICs resisted both 5-fluorouracil/calcium folinate/oxaliplatin (FolFox) and cetuximab treatment, which are the most common therapeutic regimens used for patients with advanced or metastatic rectal cancer.Thus, the identification, expansion, and properties of R-CICs provide an ideal cellular model to further investigate tumor progression and determine therapeutic resistance in these patients.

View Article: PubMed Central - PubMed

Affiliation: 1] Institute of Digestive Surgery, West China Hospital, Sichuan University, Chengdu, People's Republic of China [2] Medical Center of Stem Cell Biology, West China Hospital, Sichuan University, Chengdu, People's Republic of China [3] Department of Gastrointestinal Surgery, West China Hospital, Sichuan University, Chengdu, People's Republic of China.

ABSTRACT
Accumulating evidence indicates that cancer-initiating cells (CICs) are responsible for cancer initiation, relapse, and metastasis. Colorectal carcinoma (CRC) is typically classified into proximal colon, distal colon, and rectal cancer. The gradual changes in CRC molecular features within the bowel may have considerable implications in colon and rectal CICs. Unfortunately, limited information is available on CICs derived from rectal cancer, although colon CICs have been described. Here we identified rectal CICs (R-CICs) that possess differentiation potential in tumors derived from patients with rectal adenocarcinoma. The R-CICs carried both CD44 and CD54 surface markers, while R-CICs and their immediate progenies carried potential epithelial-mesenchymal transition characteristics. These R-CICs generated tumors similar to their tumor of origin when injected into immunodeficient mice, differentiated into rectal epithelial cells in vitro, and were capable of self-renewal both in vitro and in vivo. More importantly, subpopulations of R-CICs resisted both 5-fluorouracil/calcium folinate/oxaliplatin (FolFox) and cetuximab treatment, which are the most common therapeutic regimens used for patients with advanced or metastatic rectal cancer. Thus, the identification, expansion, and properties of R-CICs provide an ideal cellular model to further investigate tumor progression and determine therapeutic resistance in these patients.

Show MeSH
Related in: MedlinePlus