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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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Karyotype and Fluroscent In Situ Hybridization (FISH) analysis of MPE derived tumor cells.(A) Dual color FISH analysis was done using 1p36 and 1q25 (control) probes. Representative position of the probe 1p36 (orange) and 1q25 (green) on chromosome 1. (B) Sample M-1: Abnormal hyperdiploid karyotype (83 chromosomes) and (C) with 3 copies of chromosome 1 (↑) but have 2 copies (Δ) of rearranged 1p and 1q. (D) Sample M-2: Abnormal hyperdiploid karyotype (67 chromosomes) and (E) with 4 Chromsome 1 s (↑) (3 with intact 1p/1q and 1 with 1p deletion (Δ)). (F) Sample M-3: Abnormal hyperdiploid karyotype (74 chromosomes) and (G) with 2 copies of 1p (Δ) and 6 (↑) copies of 1q (consistent with 1p deletion). (H) Sample NCI-H2122 Abnormal karyotype (58 chromosomes) and (I) with 2 copies (↑and Δ) of 1p/1q but one 1p is rearranged with additional material of unknown origin at 1p terminal region (Δ). (J) Normal deployed human fibroblast cell line GM 05399 control with two copies of 1p/1q (↑).
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pone-0073195-g005: Karyotype and Fluroscent In Situ Hybridization (FISH) analysis of MPE derived tumor cells.(A) Dual color FISH analysis was done using 1p36 and 1q25 (control) probes. Representative position of the probe 1p36 (orange) and 1q25 (green) on chromosome 1. (B) Sample M-1: Abnormal hyperdiploid karyotype (83 chromosomes) and (C) with 3 copies of chromosome 1 (↑) but have 2 copies (Δ) of rearranged 1p and 1q. (D) Sample M-2: Abnormal hyperdiploid karyotype (67 chromosomes) and (E) with 4 Chromsome 1 s (↑) (3 with intact 1p/1q and 1 with 1p deletion (Δ)). (F) Sample M-3: Abnormal hyperdiploid karyotype (74 chromosomes) and (G) with 2 copies of 1p (Δ) and 6 (↑) copies of 1q (consistent with 1p deletion). (H) Sample NCI-H2122 Abnormal karyotype (58 chromosomes) and (I) with 2 copies (↑and Δ) of 1p/1q but one 1p is rearranged with additional material of unknown origin at 1p terminal region (Δ). (J) Normal deployed human fibroblast cell line GM 05399 control with two copies of 1p/1q (↑).

Mentions: Abnormal chromosomal numbers, and both hyper- and aneuploidy are common in lung cancer. It is not clear whether such chromosomal changes are associated with the tumorigenic potential of cancer cells. To investigate a possible association, karyotype analysis was performed on the three MPE samples. Normal fibroblast GM 05399 and the lung cancer cell line NCI-H2122 were used as controls to represent non tumorigenic and immortalized tumor cell models [20]–[23]. All three MPE samples M-1, M-2 and M-3 showed extensive chromosomal changes with hyperdiploid number of chromosomes 83, 67, and 74 respectively (Figure 5B, D, F). Meanwhile, the normal fibroblast contained 46 chromosomes; the cell line NCI-H2122 contained 58 chromosomes (Figure 5H). MPE cells uniformly contained translocations and deletions, and rearrangements at chromosomal region 1p, a common site of rearrangements seen in lung cancers. A FISH analysis was carried out using a 1p36 (orange) probe and a control 1q25 (green) probe to detect specific 1p changes (Figure 5A). Sample M-1 has 3 copies of chromosome 1 (↑), of which 2 copies (Δ) are rearranged at 1p and 1q (Figure 5C). The sample M-2 exhibits 4 Chromosome 1 (↑) (3 with intact 1p/1q and 1 with 1p deletion (Δ)) (Figure 5 E). The third sample M-3 has 2 copies of 1p (Δ) and 6 (↑) copies of 1q (consistent with 1p deletion) (Figure 5E).


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)

Karyotype and Fluroscent In Situ Hybridization (FISH) analysis of MPE derived tumor cells.(A) Dual color FISH analysis was done using 1p36 and 1q25 (control) probes. Representative position of the probe 1p36 (orange) and 1q25 (green) on chromosome 1. (B) Sample M-1: Abnormal hyperdiploid karyotype (83 chromosomes) and (C) with 3 copies of chromosome 1 (↑) but have 2 copies (Δ) of rearranged 1p and 1q. (D) Sample M-2: Abnormal hyperdiploid karyotype (67 chromosomes) and (E) with 4 Chromsome 1 s (↑) (3 with intact 1p/1q and 1 with 1p deletion (Δ)). (F) Sample M-3: Abnormal hyperdiploid karyotype (74 chromosomes) and (G) with 2 copies of 1p (Δ) and 6 (↑) copies of 1q (consistent with 1p deletion). (H) Sample NCI-H2122 Abnormal karyotype (58 chromosomes) and (I) with 2 copies (↑and Δ) of 1p/1q but one 1p is rearranged with additional material of unknown origin at 1p terminal region (Δ). (J) Normal deployed human fibroblast cell line GM 05399 control with two copies of 1p/1q (↑).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0073195-g005: Karyotype and Fluroscent In Situ Hybridization (FISH) analysis of MPE derived tumor cells.(A) Dual color FISH analysis was done using 1p36 and 1q25 (control) probes. Representative position of the probe 1p36 (orange) and 1q25 (green) on chromosome 1. (B) Sample M-1: Abnormal hyperdiploid karyotype (83 chromosomes) and (C) with 3 copies of chromosome 1 (↑) but have 2 copies (Δ) of rearranged 1p and 1q. (D) Sample M-2: Abnormal hyperdiploid karyotype (67 chromosomes) and (E) with 4 Chromsome 1 s (↑) (3 with intact 1p/1q and 1 with 1p deletion (Δ)). (F) Sample M-3: Abnormal hyperdiploid karyotype (74 chromosomes) and (G) with 2 copies of 1p (Δ) and 6 (↑) copies of 1q (consistent with 1p deletion). (H) Sample NCI-H2122 Abnormal karyotype (58 chromosomes) and (I) with 2 copies (↑and Δ) of 1p/1q but one 1p is rearranged with additional material of unknown origin at 1p terminal region (Δ). (J) Normal deployed human fibroblast cell line GM 05399 control with two copies of 1p/1q (↑).
Mentions: Abnormal chromosomal numbers, and both hyper- and aneuploidy are common in lung cancer. It is not clear whether such chromosomal changes are associated with the tumorigenic potential of cancer cells. To investigate a possible association, karyotype analysis was performed on the three MPE samples. Normal fibroblast GM 05399 and the lung cancer cell line NCI-H2122 were used as controls to represent non tumorigenic and immortalized tumor cell models [20]–[23]. All three MPE samples M-1, M-2 and M-3 showed extensive chromosomal changes with hyperdiploid number of chromosomes 83, 67, and 74 respectively (Figure 5B, D, F). Meanwhile, the normal fibroblast contained 46 chromosomes; the cell line NCI-H2122 contained 58 chromosomes (Figure 5H). MPE cells uniformly contained translocations and deletions, and rearrangements at chromosomal region 1p, a common site of rearrangements seen in lung cancers. A FISH analysis was carried out using a 1p36 (orange) probe and a control 1q25 (green) probe to detect specific 1p changes (Figure 5A). Sample M-1 has 3 copies of chromosome 1 (↑), of which 2 copies (Δ) are rearranged at 1p and 1q (Figure 5C). The sample M-2 exhibits 4 Chromosome 1 (↑) (3 with intact 1p/1q and 1 with 1p deletion (Δ)) (Figure 5 E). The third sample M-3 has 2 copies of 1p (Δ) and 6 (↑) copies of 1q (consistent with 1p deletion) (Figure 5E).

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