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Selective Activation of Cancer Stem Cells by Size-Specific Hyaluronan in Head and Neck Cancer.

Shiina M, Bourguignon LY - Int J Cell Biol (2015)

Bottom Line: Survival protein (cIAP-1) expression was also stimulated by 200 kDa-HA in these CSCs leading to cisplatin resistance.Furthermore, our results indicate that the anti-miR-10 inhibitor not only decreases survival protein expression, but also increases chemosensitivity of the 200 kDa-HA-treated CSCs.These findings strongly support the contention that 200 kDa-HA plays a pivotal role in miR-10 production leading to survival protein upregulation and chemoresistance in CSCs.

View Article: PubMed Central - PubMed

Affiliation: San Francisco Veterans Affairs Medical Center and Department of Medicine, University of California at San Francisco and Endocrine Unit (111N2), 4150 Clement Street, San Francisco, CA 94121, USA.

ABSTRACT
We determined that human head and neck cancer cells (HSC-3 cell line) contain a subpopulation displaying cancer stem cell (CSC) properties and are very tumorigenic. Specifically, we investigated whether different sizes of hyaluronan (HA) (e.g., 5 kDa, 20 kDa, 200 kDa, or 700 kDa-HA-sizes) play a role in regulating these CSCs. First, we observed that 200 kDa-HA (but not other sizes of HA) preferentially induces certain stem cell marker expression resulting in self-renewal and clonal formation of these cells. Further analyses indicate that 200 kDa-HA selectively stimulates the expression of a panel of microRNAs (most noticeably miR-10b) in these CSCs. Survival protein (cIAP-1) expression was also stimulated by 200 kDa-HA in these CSCs leading to cisplatin resistance. Furthermore, our results indicate that the anti-miR-10 inhibitor not only decreases survival protein expression, but also increases chemosensitivity of the 200 kDa-HA-treated CSCs. These findings strongly support the contention that 200 kDa-HA plays a pivotal role in miR-10 production leading to survival protein upregulation and chemoresistance in CSCs. Together, our findings suggest that selective activation of oncogenic signaling by certain sizes of HA (e.g., 200 kDa-HA) may be instrumental in the formation of CSC functions leading to tumor cell survival and chemoresistance in head and neck cancer progression.

No MeSH data available.


Related in: MedlinePlus

Measurement of sphere formation (a) and clone formation (b) CD44v3highALDH1high (CSC) cells. (a) Sphere formation of CD44v3highALDH1high cells [treated with no HA (A) or with 200 kDa-HA (B)] in a mixture of 5 mg/mL of matrigel (Corning) and defined medium (RPMI-1640 medium containing EGF and bFGF without serum) for 3 weeks (21 days) as described in Section 2. (b) Clone formation (differentiation) of CD44v3highALDH1high cells pretreated with no HA (A) or with 200 kDa-HA (B). Specifically, clone formation and differentiation were induced by incubating CD44v3highALDHhigh cells (dissociated from spheres treated with 200 kDa-HA or without HA for 10 days as described above). After the removal of 200 kDa-HA from the cells, these sphere-derived CD44v3highALDHhigh cells were then incubated in RPMI 1640 complete culture and 10% fetal bovine serum for ~7–10 days. After most cell clones expand to >50–100 cells, they were fixed with methanol followed by staining with crystal violet to visualize clone formation as described in Section 2.
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fig3: Measurement of sphere formation (a) and clone formation (b) CD44v3highALDH1high (CSC) cells. (a) Sphere formation of CD44v3highALDH1high cells [treated with no HA (A) or with 200 kDa-HA (B)] in a mixture of 5 mg/mL of matrigel (Corning) and defined medium (RPMI-1640 medium containing EGF and bFGF without serum) for 3 weeks (21 days) as described in Section 2. (b) Clone formation (differentiation) of CD44v3highALDH1high cells pretreated with no HA (A) or with 200 kDa-HA (B). Specifically, clone formation and differentiation were induced by incubating CD44v3highALDHhigh cells (dissociated from spheres treated with 200 kDa-HA or without HA for 10 days as described above). After the removal of 200 kDa-HA from the cells, these sphere-derived CD44v3highALDHhigh cells were then incubated in RPMI 1640 complete culture and 10% fetal bovine serum for ~7–10 days. After most cell clones expand to >50–100 cells, they were fixed with methanol followed by staining with crystal violet to visualize clone formation as described in Section 2.

Mentions: To determine whether 200 kDA-HA-treated CD44v3highALDH1high cells (overexpressing stem cell markers, Nanog, Oct4, Sox2, and KLF-4) are capable of undergoing self-renewal and long-term tumor cell growth, we assessed the ability of these tumorigenic CD44v3highALDH1high cells to grow in a “sphere forming” culture by incubating them in serum-free spheroid medium containing 200 kDa-HA (or no HA). After 14 days of incubating these cells in the serum-free medium, we observed that the 200 kDa-HA-treated CD44v3highALDH1high cells form large numbers of spheres (Figure 3(a)-(B)), ranging from 50 to 100 cells per spheroid (Figure 3(a)-(B)). In contrast, only a very small number of spheres were detected in those cells that were not treated with HA (Figure 3(a)-(B)). Therefore, it appears that sphere formation with CD44v3highALDH1high cells involves the binding of 200 kDa-HA.


Selective Activation of Cancer Stem Cells by Size-Specific Hyaluronan in Head and Neck Cancer.

Shiina M, Bourguignon LY - Int J Cell Biol (2015)

Measurement of sphere formation (a) and clone formation (b) CD44v3highALDH1high (CSC) cells. (a) Sphere formation of CD44v3highALDH1high cells [treated with no HA (A) or with 200 kDa-HA (B)] in a mixture of 5 mg/mL of matrigel (Corning) and defined medium (RPMI-1640 medium containing EGF and bFGF without serum) for 3 weeks (21 days) as described in Section 2. (b) Clone formation (differentiation) of CD44v3highALDH1high cells pretreated with no HA (A) or with 200 kDa-HA (B). Specifically, clone formation and differentiation were induced by incubating CD44v3highALDHhigh cells (dissociated from spheres treated with 200 kDa-HA or without HA for 10 days as described above). After the removal of 200 kDa-HA from the cells, these sphere-derived CD44v3highALDHhigh cells were then incubated in RPMI 1640 complete culture and 10% fetal bovine serum for ~7–10 days. After most cell clones expand to >50–100 cells, they were fixed with methanol followed by staining with crystal violet to visualize clone formation as described in Section 2.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4581563&req=5

fig3: Measurement of sphere formation (a) and clone formation (b) CD44v3highALDH1high (CSC) cells. (a) Sphere formation of CD44v3highALDH1high cells [treated with no HA (A) or with 200 kDa-HA (B)] in a mixture of 5 mg/mL of matrigel (Corning) and defined medium (RPMI-1640 medium containing EGF and bFGF without serum) for 3 weeks (21 days) as described in Section 2. (b) Clone formation (differentiation) of CD44v3highALDH1high cells pretreated with no HA (A) or with 200 kDa-HA (B). Specifically, clone formation and differentiation were induced by incubating CD44v3highALDHhigh cells (dissociated from spheres treated with 200 kDa-HA or without HA for 10 days as described above). After the removal of 200 kDa-HA from the cells, these sphere-derived CD44v3highALDHhigh cells were then incubated in RPMI 1640 complete culture and 10% fetal bovine serum for ~7–10 days. After most cell clones expand to >50–100 cells, they were fixed with methanol followed by staining with crystal violet to visualize clone formation as described in Section 2.
Mentions: To determine whether 200 kDA-HA-treated CD44v3highALDH1high cells (overexpressing stem cell markers, Nanog, Oct4, Sox2, and KLF-4) are capable of undergoing self-renewal and long-term tumor cell growth, we assessed the ability of these tumorigenic CD44v3highALDH1high cells to grow in a “sphere forming” culture by incubating them in serum-free spheroid medium containing 200 kDa-HA (or no HA). After 14 days of incubating these cells in the serum-free medium, we observed that the 200 kDa-HA-treated CD44v3highALDH1high cells form large numbers of spheres (Figure 3(a)-(B)), ranging from 50 to 100 cells per spheroid (Figure 3(a)-(B)). In contrast, only a very small number of spheres were detected in those cells that were not treated with HA (Figure 3(a)-(B)). Therefore, it appears that sphere formation with CD44v3highALDH1high cells involves the binding of 200 kDa-HA.

Bottom Line: Survival protein (cIAP-1) expression was also stimulated by 200 kDa-HA in these CSCs leading to cisplatin resistance.Furthermore, our results indicate that the anti-miR-10 inhibitor not only decreases survival protein expression, but also increases chemosensitivity of the 200 kDa-HA-treated CSCs.These findings strongly support the contention that 200 kDa-HA plays a pivotal role in miR-10 production leading to survival protein upregulation and chemoresistance in CSCs.

View Article: PubMed Central - PubMed

Affiliation: San Francisco Veterans Affairs Medical Center and Department of Medicine, University of California at San Francisco and Endocrine Unit (111N2), 4150 Clement Street, San Francisco, CA 94121, USA.

ABSTRACT
We determined that human head and neck cancer cells (HSC-3 cell line) contain a subpopulation displaying cancer stem cell (CSC) properties and are very tumorigenic. Specifically, we investigated whether different sizes of hyaluronan (HA) (e.g., 5 kDa, 20 kDa, 200 kDa, or 700 kDa-HA-sizes) play a role in regulating these CSCs. First, we observed that 200 kDa-HA (but not other sizes of HA) preferentially induces certain stem cell marker expression resulting in self-renewal and clonal formation of these cells. Further analyses indicate that 200 kDa-HA selectively stimulates the expression of a panel of microRNAs (most noticeably miR-10b) in these CSCs. Survival protein (cIAP-1) expression was also stimulated by 200 kDa-HA in these CSCs leading to cisplatin resistance. Furthermore, our results indicate that the anti-miR-10 inhibitor not only decreases survival protein expression, but also increases chemosensitivity of the 200 kDa-HA-treated CSCs. These findings strongly support the contention that 200 kDa-HA plays a pivotal role in miR-10 production leading to survival protein upregulation and chemoresistance in CSCs. Together, our findings suggest that selective activation of oncogenic signaling by certain sizes of HA (e.g., 200 kDa-HA) may be instrumental in the formation of CSC functions leading to tumor cell survival and chemoresistance in head and neck cancer progression.

No MeSH data available.


Related in: MedlinePlus