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Coexpression of CD44-positive/CD133-positive cancer stem cells and CD204-positive tumor-associated macrophages is a predictor of survival in pancreatic ductal adenocarcinoma.

Hou YC, Chao YJ, Tung HL, Wang HC, Shan YS - Cancer (2014)

Bottom Line: Expression levels of CD44/CD133 and CD204 were significantly higher in tumor tissues than in normal tissues (P < .0001).The variables associated with survival were high coexpression of CD44/CD133 (P = .000), high expression of CD204 (P = .011), and tumor grade (P = .014).Survival analysis indicated that high coexpression of CD44/CD133 and CD204 was associated significantly with shorter overall survival (P = .000) and disease-free survival (P = .003).

View Article: PubMed Central - PubMed

Affiliation: Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan.

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Expression of the tumor-associated macrophages marker CD204 is observed in tissue microarrays (TMAs) and corresponding full sections of pancreatic ductal adenocarcinoma tissue. (A) Two different TMA cores from the same tumor have a similar pattern of low CD204 expression, and (B) Two different TMA cores from the same tumor have a similar pattern of high CD204 expression. White arrows indicate CD204-positive cells (original magnification ×4 in A and B; scale bars = 200 μm). (C) Part of a slide from the donor block of the same tumor is shown at ×200 original magnification (scale bar = 50 μm), and at ×400 magnification on the right (scale bar = 20 μm). The paraffin-embedded patient tissue samples were stained with anti-CD204 antibody (green) and with 4′,6-diamidino-2-phenylindole (DAPI) for cell nuclei (blue). (D) CD204 expression was compared between pancreatic cancer specimens and their matched normal tissues (n = 11 pairs; P < .0001). Values indicate the mean ± standard error. (E) CD204 expression levels (mean ± standard error) were compared between pancreatic cancer tissues and adjacent normal tissues (P < .0001).
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fig02: Expression of the tumor-associated macrophages marker CD204 is observed in tissue microarrays (TMAs) and corresponding full sections of pancreatic ductal adenocarcinoma tissue. (A) Two different TMA cores from the same tumor have a similar pattern of low CD204 expression, and (B) Two different TMA cores from the same tumor have a similar pattern of high CD204 expression. White arrows indicate CD204-positive cells (original magnification ×4 in A and B; scale bars = 200 μm). (C) Part of a slide from the donor block of the same tumor is shown at ×200 original magnification (scale bar = 50 μm), and at ×400 magnification on the right (scale bar = 20 μm). The paraffin-embedded patient tissue samples were stained with anti-CD204 antibody (green) and with 4′,6-diamidino-2-phenylindole (DAPI) for cell nuclei (blue). (D) CD204 expression was compared between pancreatic cancer specimens and their matched normal tissues (n = 11 pairs; P < .0001). Values indicate the mean ± standard error. (E) CD204 expression levels (mean ± standard error) were compared between pancreatic cancer tissues and adjacent normal tissues (P < .0001).

Mentions: TMAs sections were double stained with antibodies against the CSCs markers CD44 and CD133 or were single stained with antibody against the TAMs marker CD204. These antibodies were also used in Western blot and immunofluorescence staining to confirm CD44, CD133, or CD204 expression in cell lines (data not shown). To verify macrophages infiltration at the tumor site, those sections were also stained with antibody against the macrophages marker CD68. Different levels of CD44 and CD133 or coexpression of CD44 and CD133 (CD44/CD133) are illustrated in Figure 1A and 1B, and different expression levels of CD204 are illustrated in Figure 2A and 2B. To validate the accuracy of our data, corresponding full sections of PDAC tumors and normal tissues were stained for these markers, as displayed in Figures 1C and 2C. CD44/CD133 expression and CD204 expression were significantly higher in tumor tissues than in their normal tissue counterparts (Figs. 1D, 1E, 2D, and 2E). The mean ± standard error expression levels of CD44 (9.71% ± 0.58%), CD133 (7.31% ± 0.44%), CD44/CD133 (2.5% ± 0.18%), CD68 (22.39% ± 0.8%), and CD204 (9.18% ± 0.49%) were used to divide patients into a high-expression group and a low-expression group. In the entire pancreatic cancer cohort, 31 of 96 patients (32.3%) had high CD44/CD133 expression, and 65 of 96 patients (67.7%) had low CD44/CD133 expression; whereas 39 of 96 patients (40.6%) had high CD204 expression, and 57 of 96 patients (59.4%) had low CD204 expression (Table 1).


Coexpression of CD44-positive/CD133-positive cancer stem cells and CD204-positive tumor-associated macrophages is a predictor of survival in pancreatic ductal adenocarcinoma.

Hou YC, Chao YJ, Tung HL, Wang HC, Shan YS - Cancer (2014)

Expression of the tumor-associated macrophages marker CD204 is observed in tissue microarrays (TMAs) and corresponding full sections of pancreatic ductal adenocarcinoma tissue. (A) Two different TMA cores from the same tumor have a similar pattern of low CD204 expression, and (B) Two different TMA cores from the same tumor have a similar pattern of high CD204 expression. White arrows indicate CD204-positive cells (original magnification ×4 in A and B; scale bars = 200 μm). (C) Part of a slide from the donor block of the same tumor is shown at ×200 original magnification (scale bar = 50 μm), and at ×400 magnification on the right (scale bar = 20 μm). The paraffin-embedded patient tissue samples were stained with anti-CD204 antibody (green) and with 4′,6-diamidino-2-phenylindole (DAPI) for cell nuclei (blue). (D) CD204 expression was compared between pancreatic cancer specimens and their matched normal tissues (n = 11 pairs; P < .0001). Values indicate the mean ± standard error. (E) CD204 expression levels (mean ± standard error) were compared between pancreatic cancer tissues and adjacent normal tissues (P < .0001).
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fig02: Expression of the tumor-associated macrophages marker CD204 is observed in tissue microarrays (TMAs) and corresponding full sections of pancreatic ductal adenocarcinoma tissue. (A) Two different TMA cores from the same tumor have a similar pattern of low CD204 expression, and (B) Two different TMA cores from the same tumor have a similar pattern of high CD204 expression. White arrows indicate CD204-positive cells (original magnification ×4 in A and B; scale bars = 200 μm). (C) Part of a slide from the donor block of the same tumor is shown at ×200 original magnification (scale bar = 50 μm), and at ×400 magnification on the right (scale bar = 20 μm). The paraffin-embedded patient tissue samples were stained with anti-CD204 antibody (green) and with 4′,6-diamidino-2-phenylindole (DAPI) for cell nuclei (blue). (D) CD204 expression was compared between pancreatic cancer specimens and their matched normal tissues (n = 11 pairs; P < .0001). Values indicate the mean ± standard error. (E) CD204 expression levels (mean ± standard error) were compared between pancreatic cancer tissues and adjacent normal tissues (P < .0001).
Mentions: TMAs sections were double stained with antibodies against the CSCs markers CD44 and CD133 or were single stained with antibody against the TAMs marker CD204. These antibodies were also used in Western blot and immunofluorescence staining to confirm CD44, CD133, or CD204 expression in cell lines (data not shown). To verify macrophages infiltration at the tumor site, those sections were also stained with antibody against the macrophages marker CD68. Different levels of CD44 and CD133 or coexpression of CD44 and CD133 (CD44/CD133) are illustrated in Figure 1A and 1B, and different expression levels of CD204 are illustrated in Figure 2A and 2B. To validate the accuracy of our data, corresponding full sections of PDAC tumors and normal tissues were stained for these markers, as displayed in Figures 1C and 2C. CD44/CD133 expression and CD204 expression were significantly higher in tumor tissues than in their normal tissue counterparts (Figs. 1D, 1E, 2D, and 2E). The mean ± standard error expression levels of CD44 (9.71% ± 0.58%), CD133 (7.31% ± 0.44%), CD44/CD133 (2.5% ± 0.18%), CD68 (22.39% ± 0.8%), and CD204 (9.18% ± 0.49%) were used to divide patients into a high-expression group and a low-expression group. In the entire pancreatic cancer cohort, 31 of 96 patients (32.3%) had high CD44/CD133 expression, and 65 of 96 patients (67.7%) had low CD44/CD133 expression; whereas 39 of 96 patients (40.6%) had high CD204 expression, and 57 of 96 patients (59.4%) had low CD204 expression (Table 1).

Bottom Line: Expression levels of CD44/CD133 and CD204 were significantly higher in tumor tissues than in normal tissues (P < .0001).The variables associated with survival were high coexpression of CD44/CD133 (P = .000), high expression of CD204 (P = .011), and tumor grade (P = .014).Survival analysis indicated that high coexpression of CD44/CD133 and CD204 was associated significantly with shorter overall survival (P = .000) and disease-free survival (P = .003).

View Article: PubMed Central - PubMed

Affiliation: Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan.

Show MeSH
Related in: MedlinePlus