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The CD44(high) tumorigenic subsets in lung cancer biospecimens are enriched for low miR-34a expression.

Basak SK, Veena MS, Oh S, Lai C, Vangala S, Elashoff D, Fishbein MC, Sharma S, Rao NP, Rao D, Phan R, Srivatsan ES, Batra RK - PLoS ONE (2013)

Bottom Line: Progression can be associated with emergence of cells that exhibit high phenotypic plasticity (including "de-differentiation" to primitive developmental states), and aggressive behavioral properties (including high tumorigenic potentials).The colony forming efficiency of CD44(hi) cells, characteristic property of CSC, can be inhibited by mir-34a replacement in these samples.In addition the highly tumorigenic CD44(hi) cells are enriched for cells in the G2 phase of cell cycle.

View Article: PubMed Central - PubMed

Affiliation: Wadsworth Stem Cell Institute, Veterans Affairs Greater Los Angeles Healthcare System (VAGLAHS), Los Angeles, California, United States of America ; Department of Medicine, David Geffen School of Medicine at UCLA, Los Angeles, California, United States of America.

ABSTRACT
Cellular heterogeneity is an integral part of cancer development and progression. Progression can be associated with emergence of cells that exhibit high phenotypic plasticity (including "de-differentiation" to primitive developmental states), and aggressive behavioral properties (including high tumorigenic potentials). We observed that many biomarkers that are used to identify Cancer Stem Cells (CSC) can label cell subsets in an advanced clinical stage of lung cancer (malignant pleural effusions, or MPE). Thus, CSC-biomarkers may be useful for live sorting functionally distinct cell subsets from individual tumors, which may enable investigators to hone in on the molecular basis for functional heterogeneity. We demonstrate that the CD44(hi) (CD44-high) cancer cell subsets display higher clonal, colony forming potential than CD44(lo) cells (n=3) and are also tumorigenic (n=2/2) when transplanted in mouse xenograft model. The CD44(hi) subsets express different levels of embryonal (de-differentiation) markers or chromatin regulators. In archived lung cancer tissues, ALDH markers co-localize more with CD44 in squamous cell carcinoma (n=5/7) than Adeno Carcinoma (n=1/12). MPE cancer cells and a lung cancer cell line (NCI-H-2122) exhibit chromosomal abnormalities and 1p36 deletion (n=3/3). Since miR-34a maps to the 1p36 deletion site, low miR-34a expression levels were detected in these cells. The colony forming efficiency of CD44(hi) cells, characteristic property of CSC, can be inhibited by mir-34a replacement in these samples. In addition the highly tumorigenic CD44(hi) cells are enriched for cells in the G2 phase of cell cycle.

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Immunohistological study of tumors generated by CD44hi cell population in mouse, human squamous cell carcinomas (SCCs), human alveolar and human bronchiolar tissues.The photomicrograph A, B, C and D represent the tumors derived from Sample M-1 and E, F, G and H represent tumor derived from sample M-2 in NOD/SCID (IL2rγ) mice. The following stains are represented: H&E staining (A and E), immunohistochemistry for CD44 expression (B and F), ALDH expression pattern (C and G) and the immunohistochemistry for dual CD44 and ALDH staining (D and H). Representative human two lung squamous cell carcinoma (SCC) tissue samples (I, J, K and L, M, N) were stained by H&E (I), CD44 (J and L), ALDH (K and M) and immunohistochemical staining for dual markers CD44 and ALDH (N). Human alveolar tissue sections were stained for CD44 (O), ALDH (P) and dual markers CD44 and ALDH (Q). Human-bronchiolar tissue sample was stained for CD44(R), ALDH (S) and dual markers CD44 and ALDH (T).
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pone-0073195-g004: Immunohistological study of tumors generated by CD44hi cell population in mouse, human squamous cell carcinomas (SCCs), human alveolar and human bronchiolar tissues.The photomicrograph A, B, C and D represent the tumors derived from Sample M-1 and E, F, G and H represent tumor derived from sample M-2 in NOD/SCID (IL2rγ) mice. The following stains are represented: H&E staining (A and E), immunohistochemistry for CD44 expression (B and F), ALDH expression pattern (C and G) and the immunohistochemistry for dual CD44 and ALDH staining (D and H). Representative human two lung squamous cell carcinoma (SCC) tissue samples (I, J, K and L, M, N) were stained by H&E (I), CD44 (J and L), ALDH (K and M) and immunohistochemical staining for dual markers CD44 and ALDH (N). Human alveolar tissue sections were stained for CD44 (O), ALDH (P) and dual markers CD44 and ALDH (Q). Human-bronchiolar tissue sample was stained for CD44(R), ALDH (S) and dual markers CD44 and ALDH (T).

Mentions: CD44hi cells in MPE primary cultures contain cell fractions with high ALDH expression (i.e., the CD44hi/ALDHhi surface phenotype) [5]. We extend that observation to prospectively collected biospecimens. Using immunohistochemistry, we observe variable expression of CD44 and ALDH markers in the mouse xenograft tumors generated from CD44hi cells. Pathological and marker expression patterns in xenografts compare favorably to archived human lung cancer, and to tumor-adjacent human normal alveolar and normal human bronchiolar tissues. H&E sections of M-1 and M-2 CD44hi xenografts (Figure 4 A–H) corroborate the original pathological diagnoses of large cell lung cancer and lung SCC respectively (Figure 4 I–N). Consistent with flow cytometry data, CD44 labeling is evident on the majority of cells. However, intra-tumoral variation of CD44 expression is clearly evident (Figure 4 B, F), again consistent with the flow cytometry profile. Similarly when the xenograft sections are labeled for ALDH expression, some cells show higher expression than other tumor cell populations (Figure 4 C, G). When co-expression of CD44 and ALDH is examined by dual marker staining of xenograft tumor sections, there is tumor to tumor variability in the co-localization of these markers (Figure 4 D, H).


The CD44(high) tumorigenic subsets in lung cancer biospecimens are enriched for low miR-34a expression.

Basak SK, Veena MS, Oh S, Lai C, Vangala S, Elashoff D, Fishbein MC, Sharma S, Rao NP, Rao D, Phan R, Srivatsan ES, Batra RK - PLoS ONE (2013)

Immunohistological study of tumors generated by CD44hi cell population in mouse, human squamous cell carcinomas (SCCs), human alveolar and human bronchiolar tissues.The photomicrograph A, B, C and D represent the tumors derived from Sample M-1 and E, F, G and H represent tumor derived from sample M-2 in NOD/SCID (IL2rγ) mice. The following stains are represented: H&E staining (A and E), immunohistochemistry for CD44 expression (B and F), ALDH expression pattern (C and G) and the immunohistochemistry for dual CD44 and ALDH staining (D and H). Representative human two lung squamous cell carcinoma (SCC) tissue samples (I, J, K and L, M, N) were stained by H&E (I), CD44 (J and L), ALDH (K and M) and immunohistochemical staining for dual markers CD44 and ALDH (N). Human alveolar tissue sections were stained for CD44 (O), ALDH (P) and dual markers CD44 and ALDH (Q). Human-bronchiolar tissue sample was stained for CD44(R), ALDH (S) and dual markers CD44 and ALDH (T).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3760902&req=5

pone-0073195-g004: Immunohistological study of tumors generated by CD44hi cell population in mouse, human squamous cell carcinomas (SCCs), human alveolar and human bronchiolar tissues.The photomicrograph A, B, C and D represent the tumors derived from Sample M-1 and E, F, G and H represent tumor derived from sample M-2 in NOD/SCID (IL2rγ) mice. The following stains are represented: H&E staining (A and E), immunohistochemistry for CD44 expression (B and F), ALDH expression pattern (C and G) and the immunohistochemistry for dual CD44 and ALDH staining (D and H). Representative human two lung squamous cell carcinoma (SCC) tissue samples (I, J, K and L, M, N) were stained by H&E (I), CD44 (J and L), ALDH (K and M) and immunohistochemical staining for dual markers CD44 and ALDH (N). Human alveolar tissue sections were stained for CD44 (O), ALDH (P) and dual markers CD44 and ALDH (Q). Human-bronchiolar tissue sample was stained for CD44(R), ALDH (S) and dual markers CD44 and ALDH (T).
Mentions: CD44hi cells in MPE primary cultures contain cell fractions with high ALDH expression (i.e., the CD44hi/ALDHhi surface phenotype) [5]. We extend that observation to prospectively collected biospecimens. Using immunohistochemistry, we observe variable expression of CD44 and ALDH markers in the mouse xenograft tumors generated from CD44hi cells. Pathological and marker expression patterns in xenografts compare favorably to archived human lung cancer, and to tumor-adjacent human normal alveolar and normal human bronchiolar tissues. H&E sections of M-1 and M-2 CD44hi xenografts (Figure 4 A–H) corroborate the original pathological diagnoses of large cell lung cancer and lung SCC respectively (Figure 4 I–N). Consistent with flow cytometry data, CD44 labeling is evident on the majority of cells. However, intra-tumoral variation of CD44 expression is clearly evident (Figure 4 B, F), again consistent with the flow cytometry profile. Similarly when the xenograft sections are labeled for ALDH expression, some cells show higher expression than other tumor cell populations (Figure 4 C, G). When co-expression of CD44 and ALDH is examined by dual marker staining of xenograft tumor sections, there is tumor to tumor variability in the co-localization of these markers (Figure 4 D, H).

Bottom Line: Progression can be associated with emergence of cells that exhibit high phenotypic plasticity (including "de-differentiation" to primitive developmental states), and aggressive behavioral properties (including high tumorigenic potentials).The colony forming efficiency of CD44(hi) cells, characteristic property of CSC, can be inhibited by mir-34a replacement in these samples.In addition the highly tumorigenic CD44(hi) cells are enriched for cells in the G2 phase of cell cycle.

View Article: PubMed Central - PubMed

Affiliation: Wadsworth Stem Cell Institute, Veterans Affairs Greater Los Angeles Healthcare System (VAGLAHS), Los Angeles, California, United States of America ; Department of Medicine, David Geffen School of Medicine at UCLA, Los Angeles, California, United States of America.

ABSTRACT
Cellular heterogeneity is an integral part of cancer development and progression. Progression can be associated with emergence of cells that exhibit high phenotypic plasticity (including "de-differentiation" to primitive developmental states), and aggressive behavioral properties (including high tumorigenic potentials). We observed that many biomarkers that are used to identify Cancer Stem Cells (CSC) can label cell subsets in an advanced clinical stage of lung cancer (malignant pleural effusions, or MPE). Thus, CSC-biomarkers may be useful for live sorting functionally distinct cell subsets from individual tumors, which may enable investigators to hone in on the molecular basis for functional heterogeneity. We demonstrate that the CD44(hi) (CD44-high) cancer cell subsets display higher clonal, colony forming potential than CD44(lo) cells (n=3) and are also tumorigenic (n=2/2) when transplanted in mouse xenograft model. The CD44(hi) subsets express different levels of embryonal (de-differentiation) markers or chromatin regulators. In archived lung cancer tissues, ALDH markers co-localize more with CD44 in squamous cell carcinoma (n=5/7) than Adeno Carcinoma (n=1/12). MPE cancer cells and a lung cancer cell line (NCI-H-2122) exhibit chromosomal abnormalities and 1p36 deletion (n=3/3). Since miR-34a maps to the 1p36 deletion site, low miR-34a expression levels were detected in these cells. The colony forming efficiency of CD44(hi) cells, characteristic property of CSC, can be inhibited by mir-34a replacement in these samples. In addition the highly tumorigenic CD44(hi) cells are enriched for cells in the G2 phase of cell cycle.

Show MeSH
Related in: MedlinePlus