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Immunohistochemical detection of cancer stem cell related markers CD44 and CD133 in metastatic colorectal cancer patients.

Pitule P, Cedikova M, Daum O, Vojtisek J, Vycital O, Hosek P, Treska V, Hes O, Kralickova M, Liska V - Biomed Res Int (2014)

Bottom Line: Samples were stained by antibodies against CD44 and CD133.Patients with higher level of CD133 staining in CRC had longer disease free interval (Cox-Mantel P = 0.0244), whereas we found no relation between CD44 expression and overall survival or disease free interval.CD133 expression in CRC and CLM differed based on CRC grading; in case of CD44 we found differences in staining intensity in individual stages of tumor lymph node invasion.

View Article: PubMed Central - PubMed

Affiliation: Biomedical Center, Faculty of Medicine in Pilsen, Charles University in Prague, Husova 3, 306 05 Pilsen, Czech Republic ; Department of Histology and Embryology, Faculty of Medicine in Pilsen, Charles University in Prague, Karlovarsk√° 48, 301 00 Pilsen, Czech Republic.

ABSTRACT

Aim: The goal of this study was to semiquantitatively detect presence of cancer stem cells markers CD44 and CD133 in immunohistochemically stained paired samples of colorectal cancer (CRC) and colorectal liver metastases (CLM). Level of staining intensity was compared to clinical and pathological characteristics of tumors with the aim to identify impact of CD44 or CD133 expression on tumor behavior.

Patients and methods: Formalin fixed paraffin embedded samples from 94 patients with colorectal tumor and liver metastases were collected at Sikl's Department of Pathology. Samples were stained by antibodies against CD44 and CD133. Presence and intensity of staining was assessed semiquantitatively by three trained researchers.

Results: Patients with higher level of CD133 staining in CRC had longer disease free interval (Cox-Mantel P = 0.0244), whereas we found no relation between CD44 expression and overall survival or disease free interval. CD133 expression in CRC and CLM differed based on CRC grading; in case of CD44 we found differences in staining intensity in individual stages of tumor lymph node invasion.

Conclusion: Effect of cancer stem cell markers on prognosis of colorectal cancer can vary depending on pathological classification of tumor, and we have shown that CD133, generally considered to be a negative marker, can bear also clinically positive prognostic information in group of patients with colorectal liver metastases.

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Expression of CD133. (a, b) Samples negative for CD133 in the lumen of tumor glands, (c, d) samples with positive CD133 staining on the apical portions of tumor cells. Magnification 200x (a, c) and 400x (b, d) of the same samples.
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fig1: Expression of CD133. (a, b) Samples negative for CD133 in the lumen of tumor glands, (c, d) samples with positive CD133 staining on the apical portions of tumor cells. Magnification 200x (a, c) and 400x (b, d) of the same samples.

Mentions: A method based on previous study was used for analysis of slides [15]. All slides stained for CD44 and CD133 were analyzed independently by three trained researchers (Pavel Pitule, Miroslava Cedikova, and Jan Vojtisek). Tumors were localized using 10x objective and level of positivity on scale from 0 (negative) to 3 (highly positive) was assessed for CD44 staining. For CD133 we evaluated five microscopical fields using 40x objective and the percentage of CD133 positive tumor glands compared to all tumor glands in the view field were assessed. Positive staining of bile duct walls that should occur after every successful CD133 staining was used as an internal control of the staining process in case of liver metastasis samples. CLM slides with unstained bile ducts were excluded from the analysis. Examples of markers expression are summarized in Figures 1 and 2.


Immunohistochemical detection of cancer stem cell related markers CD44 and CD133 in metastatic colorectal cancer patients.

Pitule P, Cedikova M, Daum O, Vojtisek J, Vycital O, Hosek P, Treska V, Hes O, Kralickova M, Liska V - Biomed Res Int (2014)

Expression of CD133. (a, b) Samples negative for CD133 in the lumen of tumor glands, (c, d) samples with positive CD133 staining on the apical portions of tumor cells. Magnification 200x (a, c) and 400x (b, d) of the same samples.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4016925&req=5

fig1: Expression of CD133. (a, b) Samples negative for CD133 in the lumen of tumor glands, (c, d) samples with positive CD133 staining on the apical portions of tumor cells. Magnification 200x (a, c) and 400x (b, d) of the same samples.
Mentions: A method based on previous study was used for analysis of slides [15]. All slides stained for CD44 and CD133 were analyzed independently by three trained researchers (Pavel Pitule, Miroslava Cedikova, and Jan Vojtisek). Tumors were localized using 10x objective and level of positivity on scale from 0 (negative) to 3 (highly positive) was assessed for CD44 staining. For CD133 we evaluated five microscopical fields using 40x objective and the percentage of CD133 positive tumor glands compared to all tumor glands in the view field were assessed. Positive staining of bile duct walls that should occur after every successful CD133 staining was used as an internal control of the staining process in case of liver metastasis samples. CLM slides with unstained bile ducts were excluded from the analysis. Examples of markers expression are summarized in Figures 1 and 2.

Bottom Line: Samples were stained by antibodies against CD44 and CD133.Patients with higher level of CD133 staining in CRC had longer disease free interval (Cox-Mantel P = 0.0244), whereas we found no relation between CD44 expression and overall survival or disease free interval.CD133 expression in CRC and CLM differed based on CRC grading; in case of CD44 we found differences in staining intensity in individual stages of tumor lymph node invasion.

View Article: PubMed Central - PubMed

Affiliation: Biomedical Center, Faculty of Medicine in Pilsen, Charles University in Prague, Husova 3, 306 05 Pilsen, Czech Republic ; Department of Histology and Embryology, Faculty of Medicine in Pilsen, Charles University in Prague, Karlovarsk√° 48, 301 00 Pilsen, Czech Republic.

ABSTRACT

Aim: The goal of this study was to semiquantitatively detect presence of cancer stem cells markers CD44 and CD133 in immunohistochemically stained paired samples of colorectal cancer (CRC) and colorectal liver metastases (CLM). Level of staining intensity was compared to clinical and pathological characteristics of tumors with the aim to identify impact of CD44 or CD133 expression on tumor behavior.

Patients and methods: Formalin fixed paraffin embedded samples from 94 patients with colorectal tumor and liver metastases were collected at Sikl's Department of Pathology. Samples were stained by antibodies against CD44 and CD133. Presence and intensity of staining was assessed semiquantitatively by three trained researchers.

Results: Patients with higher level of CD133 staining in CRC had longer disease free interval (Cox-Mantel P = 0.0244), whereas we found no relation between CD44 expression and overall survival or disease free interval. CD133 expression in CRC and CLM differed based on CRC grading; in case of CD44 we found differences in staining intensity in individual stages of tumor lymph node invasion.

Conclusion: Effect of cancer stem cell markers on prognosis of colorectal cancer can vary depending on pathological classification of tumor, and we have shown that CD133, generally considered to be a negative marker, can bear also clinically positive prognostic information in group of patients with colorectal liver metastases.

Show MeSH
Related in: MedlinePlus