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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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Cell cycle parameters of CD44hi and CD44lo cell populations derived from primary cultures of MPE tumors.Representative FACS analysis of CD44 and PI staining (i and ii) of three samples (A) M-1, (B) M-2 and (C) M-3. The samples were evaluated for their CD44hi (iii) and CD44lo (iv) and total cells (v, no gate) cell cycle and their G1, S and G2 phase analysis. (D) The results indicate that gated CD44hi population express high level of S and G2 phase (Sample M-1: S/G2∶15.75/72.61; Sample M-2: S/G2∶12.03/32.19; Sample M-3: S/G2∶6.25/15.17) than the gated CD44lo population (Sample M-1: S/G2∶4.36/1.81; Sample M-2: S/G2∶5.18/7.14; Sample M-3: S/G2∶3.37/3.32) respectively.
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pone-0073195-g007: Cell cycle parameters of CD44hi and CD44lo cell populations derived from primary cultures of MPE tumors.Representative FACS analysis of CD44 and PI staining (i and ii) of three samples (A) M-1, (B) M-2 and (C) M-3. The samples were evaluated for their CD44hi (iii) and CD44lo (iv) and total cells (v, no gate) cell cycle and their G1, S and G2 phase analysis. (D) The results indicate that gated CD44hi population express high level of S and G2 phase (Sample M-1: S/G2∶15.75/72.61; Sample M-2: S/G2∶12.03/32.19; Sample M-3: S/G2∶6.25/15.17) than the gated CD44lo population (Sample M-1: S/G2∶4.36/1.81; Sample M-2: S/G2∶5.18/7.14; Sample M-3: S/G2∶3.37/3.32) respectively.

Mentions: Samples (A) M-1, (B) M-2 and (C) M-3 were stained for CD44 and PI and then first gated with PI staining pattern (Figure 7i) and then back-gated for CD44 (CD44-FITC/FL-1) and PI (FL-2A) (Figure 7ii). The panels iii, iv and v of figure 7 represent histogram of cell cycle stages of CD44hi and CD44lo gated cells (5–10% of total cells) and un-gated total cell population respectively. Figure 7D represents the population at different cell cycle stages G1 (M1), G2 (M2) and S (M3) stages. The CD44hi cells of sample (A) M-1 shows that S and G2 phases are 15.75% and 72.61% of the gated cell population respectively (Figure 7A iii). CD44lo cells of the same sample represent 4.36% (S) and 1.81% (G2) of gated cell population (Figure 7A iv). Thus CD44hi cells are enriched 40-fold for cells in the G2 phase than the CD44lo cells.


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)

Cell cycle parameters of CD44hi and CD44lo cell populations derived from primary cultures of MPE tumors.Representative FACS analysis of CD44 and PI staining (i and ii) of three samples (A) M-1, (B) M-2 and (C) M-3. The samples were evaluated for their CD44hi (iii) and CD44lo (iv) and total cells (v, no gate) cell cycle and their G1, S and G2 phase analysis. (D) The results indicate that gated CD44hi population express high level of S and G2 phase (Sample M-1: S/G2∶15.75/72.61; Sample M-2: S/G2∶12.03/32.19; Sample M-3: S/G2∶6.25/15.17) than the gated CD44lo population (Sample M-1: S/G2∶4.36/1.81; Sample M-2: S/G2∶5.18/7.14; Sample M-3: S/G2∶3.37/3.32) respectively.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0073195-g007: Cell cycle parameters of CD44hi and CD44lo cell populations derived from primary cultures of MPE tumors.Representative FACS analysis of CD44 and PI staining (i and ii) of three samples (A) M-1, (B) M-2 and (C) M-3. The samples were evaluated for their CD44hi (iii) and CD44lo (iv) and total cells (v, no gate) cell cycle and their G1, S and G2 phase analysis. (D) The results indicate that gated CD44hi population express high level of S and G2 phase (Sample M-1: S/G2∶15.75/72.61; Sample M-2: S/G2∶12.03/32.19; Sample M-3: S/G2∶6.25/15.17) than the gated CD44lo population (Sample M-1: S/G2∶4.36/1.81; Sample M-2: S/G2∶5.18/7.14; Sample M-3: S/G2∶3.37/3.32) respectively.
Mentions: Samples (A) M-1, (B) M-2 and (C) M-3 were stained for CD44 and PI and then first gated with PI staining pattern (Figure 7i) and then back-gated for CD44 (CD44-FITC/FL-1) and PI (FL-2A) (Figure 7ii). The panels iii, iv and v of figure 7 represent histogram of cell cycle stages of CD44hi and CD44lo gated cells (5–10% of total cells) and un-gated total cell population respectively. Figure 7D represents the population at different cell cycle stages G1 (M1), G2 (M2) and S (M3) stages. The CD44hi cells of sample (A) M-1 shows that S and G2 phases are 15.75% and 72.61% of the gated cell population respectively (Figure 7A iii). CD44lo cells of the same sample represent 4.36% (S) and 1.81% (G2) of gated cell population (Figure 7A iv). Thus CD44hi cells are enriched 40-fold for cells in the G2 phase than the CD44lo cells.

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