Limits...
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.

Show MeSH

Related in: MedlinePlus

CD44+CD54+ cells derived from rectospheres have the strongest tumorigenicity among the four cellular subpopulations. (a) Tumor-bearing mice derived from 100, 500, 1000, and 10 000 CD44+CD54+ rectal cancer cells and excised subcutaneous tumors. One representative experiment of three different tumors is shown. (b) Size of xenografts of CD44+CD54+ and CD44+CD54− derived from rectospheres. Data are mean tumor size±S.D. of 3–5 tumors per group derived from three separate patients (patients 9, 11, and 15). (c) Hematoxylin and eosin analysis of a human rectal cancer section from the original human tumor and corresponding xenografts obtained after injection of sphere cells. Bars=100 μM. Second and third xenografts: tumors were obtained from the first and second transplantation, respectively. One representative experiment of two different tumors is shown. (d) Immunohistochemical analysis of relative differentiation and putative R-CIC markers in tissue derived from original human tumor (left panel) and sphere-derived first xenografts (right panel). One representative experiment of two different tumors is shown. Bars=100 μM. (e) Size of xenografts of CD44+CD54+ and CD44+CD54− sorted from three rectal cancer tissues. Data are mean tumor size±S.D. of 3–5 tumors per group derived from three separate patients (**P<0.01, *P<0.05; patients 31, 32, and 33)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3824647&req=5

fig4: CD44+CD54+ cells derived from rectospheres have the strongest tumorigenicity among the four cellular subpopulations. (a) Tumor-bearing mice derived from 100, 500, 1000, and 10 000 CD44+CD54+ rectal cancer cells and excised subcutaneous tumors. One representative experiment of three different tumors is shown. (b) Size of xenografts of CD44+CD54+ and CD44+CD54− derived from rectospheres. Data are mean tumor size±S.D. of 3–5 tumors per group derived from three separate patients (patients 9, 11, and 15). (c) Hematoxylin and eosin analysis of a human rectal cancer section from the original human tumor and corresponding xenografts obtained after injection of sphere cells. Bars=100 μM. Second and third xenografts: tumors were obtained from the first and second transplantation, respectively. One representative experiment of two different tumors is shown. (d) Immunohistochemical analysis of relative differentiation and putative R-CIC markers in tissue derived from original human tumor (left panel) and sphere-derived first xenografts (right panel). One representative experiment of two different tumors is shown. Bars=100 μM. (e) Size of xenografts of CD44+CD54+ and CD44+CD54− sorted from three rectal cancer tissues. Data are mean tumor size±S.D. of 3–5 tumors per group derived from three separate patients (**P<0.01, *P<0.05; patients 31, 32, and 33)

Mentions: As CD44+CD54+ cells carry self-renewal ability in vitro, we examined whether double-positive cells possessed self-renewal ability in vivo through serial transplantations.7 First, the engraftment rate of different cellular subpopulations and different number of cells (100, 500, 1000, and 10 000 cells per mouse) was tested. We subcutaneously injected the indicated number of cells into nude mice and found that injection with as few as 100 purified CD44+CD54+ cells resulted in tumor formation after 4 weeks (Figures 4a and b). In contrast, one in five samples of 10 000 CD44+CD54−-injected mice formed tumors (Figure 4b). The other cellular subpopulations (CD44−CD54+ and CD44−CD54−) did not give rise to any xenotransplant tumors (Table 1). To determine whether the xenotransplant tumors initiated from CD44+CD54+ cells were serially transplantable, double-positive cell-generated tumor masses were harvested when the tumor diameters reached 1 cm and then transplanted again into nude mice (100 cells per mouse). We found that these cells ultimately generated tumors in secondary and tertiary recipients (Table 1). Hematoxylin and eosin staining showed xenograft tumors shared typical rectal cancer morphological features that were observed in the original tumor tissues surgically removed from human patients (Figure 4c). The immunostaining patterns of xenografts were also highly similar to the original human tumors (Figure 4d).14


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+ cells derived from rectospheres have the strongest tumorigenicity among the four cellular subpopulations. (a) Tumor-bearing mice derived from 100, 500, 1000, and 10 000 CD44+CD54+ rectal cancer cells and excised subcutaneous tumors. One representative experiment of three different tumors is shown. (b) Size of xenografts of CD44+CD54+ and CD44+CD54− derived from rectospheres. Data are mean tumor size±S.D. of 3–5 tumors per group derived from three separate patients (patients 9, 11, and 15). (c) Hematoxylin and eosin analysis of a human rectal cancer section from the original human tumor and corresponding xenografts obtained after injection of sphere cells. Bars=100 μM. Second and third xenografts: tumors were obtained from the first and second transplantation, respectively. One representative experiment of two different tumors is shown. (d) Immunohistochemical analysis of relative differentiation and putative R-CIC markers in tissue derived from original human tumor (left panel) and sphere-derived first xenografts (right panel). One representative experiment of two different tumors is shown. Bars=100 μM. (e) Size of xenografts of CD44+CD54+ and CD44+CD54− sorted from three rectal cancer tissues. Data are mean tumor size±S.D. of 3–5 tumors per group derived from three separate patients (**P<0.01, *P<0.05; patients 31, 32, and 33)
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig4: CD44+CD54+ cells derived from rectospheres have the strongest tumorigenicity among the four cellular subpopulations. (a) Tumor-bearing mice derived from 100, 500, 1000, and 10 000 CD44+CD54+ rectal cancer cells and excised subcutaneous tumors. One representative experiment of three different tumors is shown. (b) Size of xenografts of CD44+CD54+ and CD44+CD54− derived from rectospheres. Data are mean tumor size±S.D. of 3–5 tumors per group derived from three separate patients (patients 9, 11, and 15). (c) Hematoxylin and eosin analysis of a human rectal cancer section from the original human tumor and corresponding xenografts obtained after injection of sphere cells. Bars=100 μM. Second and third xenografts: tumors were obtained from the first and second transplantation, respectively. One representative experiment of two different tumors is shown. (d) Immunohistochemical analysis of relative differentiation and putative R-CIC markers in tissue derived from original human tumor (left panel) and sphere-derived first xenografts (right panel). One representative experiment of two different tumors is shown. Bars=100 μM. (e) Size of xenografts of CD44+CD54+ and CD44+CD54− sorted from three rectal cancer tissues. Data are mean tumor size±S.D. of 3–5 tumors per group derived from three separate patients (**P<0.01, *P<0.05; patients 31, 32, and 33)
Mentions: As CD44+CD54+ cells carry self-renewal ability in vitro, we examined whether double-positive cells possessed self-renewal ability in vivo through serial transplantations.7 First, the engraftment rate of different cellular subpopulations and different number of cells (100, 500, 1000, and 10 000 cells per mouse) was tested. We subcutaneously injected the indicated number of cells into nude mice and found that injection with as few as 100 purified CD44+CD54+ cells resulted in tumor formation after 4 weeks (Figures 4a and b). In contrast, one in five samples of 10 000 CD44+CD54−-injected mice formed tumors (Figure 4b). The other cellular subpopulations (CD44−CD54+ and CD44−CD54−) did not give rise to any xenotransplant tumors (Table 1). To determine whether the xenotransplant tumors initiated from CD44+CD54+ cells were serially transplantable, double-positive cell-generated tumor masses were harvested when the tumor diameters reached 1 cm and then transplanted again into nude mice (100 cells per mouse). We found that these cells ultimately generated tumors in secondary and tertiary recipients (Table 1). Hematoxylin and eosin staining showed xenograft tumors shared typical rectal cancer morphological features that were observed in the original tumor tissues surgically removed from human patients (Figure 4c). The immunostaining patterns of xenografts were also highly similar to the original human tumors (Figure 4d).14

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