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Melanoma cells influence the differentiation pattern of human epidermal keratinocytes.

Kodet O, Lacina L, Krejčí E, Dvořánková B, Grim M, Štork J, Kodetová D, Vlček Č, Šáchová J, Kolář M, Strnad H, Smetana K - Mol. Cancer (2015)

Bottom Line: Data were compared to results of transcriptome profiling of in vitro models, in which HPK were co-cultured with MC, normal human melanocytes, and NCSC, respectively.Differentially expressed candidate genes were verified by RT-qPCR.We conclude that the MC are able to influence locally the differentiation pattern of keratinocytes in vivo as well as in vitro.

View Article: PubMed Central - PubMed

Affiliation: 1st Faculty of Medicine, Institute of Anatomy, Charles University in Prague, U Nemocnice 3, CZ-12800 Prague, Czech Republic. strnad@img.cas.cz.

ABSTRACT

Background: Nodular melanoma is one of the most life threatening tumors with still poor therapeutic outcome. Similarly to other tumors, permissive microenvironment is essential for melanoma progression. Features of this microenvironment are arising from molecular crosstalk between the melanoma cells (MC) and the surrounding cell populations in the context of skin tissue. Here, we study the effect of melanoma cells on human primary keratinocytes (HPK). Presence of MC is as an important modulator of the tumor microenvironment and we compare it to the effect of nonmalignant lowly differentiated cells also originating from neural crest (NCSC).

Methods: Comparative morphometrical and immunohistochemical analysis of epidermis surrounding nodular melanoma (n = 100) was performed. Data were compared to results of transcriptome profiling of in vitro models, in which HPK were co-cultured with MC, normal human melanocytes, and NCSC, respectively. Differentially expressed candidate genes were verified by RT-qPCR. Biological activity of candidate proteins was assessed on cultured HPK.

Results: Epidermis surrounding nodular melanoma exhibits hyperplastic features in 90% of cases. This hyperplastic region exhibits aberrant suprabasal expression of keratin 14 accompanied by loss of keratin 10. We observe that MC and NCSC are able to increase expression of keratins 8, 14, 19, and vimentin in the co-cultured HPK. This in vitro finding partially correlates with pseudoepitheliomatous hyperplasia observed in melanoma biopsies. We provide evidence of FGF-2, CXCL-1, IL-8, and VEGF-A participation in the activity of melanoma cells on keratinocytes.

Conclusion: We conclude that the MC are able to influence locally the differentiation pattern of keratinocytes in vivo as well as in vitro. This interaction further highlights the role of intercellular interactions in melanoma. The reciprocal role of activated keratinocytes on biology of melanoma cells shall be verified in the future.

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Detection of keratin 14 (K14, A-D, F), protein S100 (B), keratin 10 (K10, E, G), and Ki67 (H, I). K14 is expressed only in basal layer in normal epidermis (A). In junctional naevi, the S100 positive naevus cells (green signal, marked by asterisk) are surrounded by K14 positive basal and suprabasal keratinocytes (B). Epidermis surrounding nodular melanoma reveals pseudoepitheliomatous hyperplasia with high expression of K14 (C), however, proliferation marker Ki67 is negative here (H). At the margin of surgical resecate (MSR) is K14 expressed in all suprabasal layers (D), this aberrant expression is accompanied by loss of differentiation marker K10 (E) and high proliferation revealed by the presence of Ki67 (I). Similarly, epidermis overlaying dermal metastasis of melanoma is K14 positive even suprabasally (F); expression of K10 is dislocated more superficially (G). The epithelium in melanoma resecates reaches its thickest point on the periphery of the tumor margin (J). The tumor center is covered by the epithelium of variable thickness (J). Scale bars denote 25 μm.
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Fig1: Detection of keratin 14 (K14, A-D, F), protein S100 (B), keratin 10 (K10, E, G), and Ki67 (H, I). K14 is expressed only in basal layer in normal epidermis (A). In junctional naevi, the S100 positive naevus cells (green signal, marked by asterisk) are surrounded by K14 positive basal and suprabasal keratinocytes (B). Epidermis surrounding nodular melanoma reveals pseudoepitheliomatous hyperplasia with high expression of K14 (C), however, proliferation marker Ki67 is negative here (H). At the margin of surgical resecate (MSR) is K14 expressed in all suprabasal layers (D), this aberrant expression is accompanied by loss of differentiation marker K10 (E) and high proliferation revealed by the presence of Ki67 (I). Similarly, epidermis overlaying dermal metastasis of melanoma is K14 positive even suprabasally (F); expression of K10 is dislocated more superficially (G). The epithelium in melanoma resecates reaches its thickest point on the periphery of the tumor margin (J). The tumor center is covered by the epithelium of variable thickness (J). Scale bars denote 25 μm.

Mentions: Significantly increased thickness of the epidermis in vicinity of tumor was observed in 90% of biopsies (more than 2 times compared to epidermal thickness at the surgical margin; mean 4.56×; median 3.84; range 0.72-12.2×, p value < 0.001 by Mann–Whitney test). The increased epidermal thickness was observed on the periphery of the tumor (Figure 1 J) without significant correlation to the value of Breslow’s depth of invasion.Figure 1


Melanoma cells influence the differentiation pattern of human epidermal keratinocytes.

Kodet O, Lacina L, Krejčí E, Dvořánková B, Grim M, Štork J, Kodetová D, Vlček Č, Šáchová J, Kolář M, Strnad H, Smetana K - Mol. Cancer (2015)

Detection of keratin 14 (K14, A-D, F), protein S100 (B), keratin 10 (K10, E, G), and Ki67 (H, I). K14 is expressed only in basal layer in normal epidermis (A). In junctional naevi, the S100 positive naevus cells (green signal, marked by asterisk) are surrounded by K14 positive basal and suprabasal keratinocytes (B). Epidermis surrounding nodular melanoma reveals pseudoepitheliomatous hyperplasia with high expression of K14 (C), however, proliferation marker Ki67 is negative here (H). At the margin of surgical resecate (MSR) is K14 expressed in all suprabasal layers (D), this aberrant expression is accompanied by loss of differentiation marker K10 (E) and high proliferation revealed by the presence of Ki67 (I). Similarly, epidermis overlaying dermal metastasis of melanoma is K14 positive even suprabasally (F); expression of K10 is dislocated more superficially (G). The epithelium in melanoma resecates reaches its thickest point on the periphery of the tumor margin (J). The tumor center is covered by the epithelium of variable thickness (J). Scale bars denote 25 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4325966&req=5

Fig1: Detection of keratin 14 (K14, A-D, F), protein S100 (B), keratin 10 (K10, E, G), and Ki67 (H, I). K14 is expressed only in basal layer in normal epidermis (A). In junctional naevi, the S100 positive naevus cells (green signal, marked by asterisk) are surrounded by K14 positive basal and suprabasal keratinocytes (B). Epidermis surrounding nodular melanoma reveals pseudoepitheliomatous hyperplasia with high expression of K14 (C), however, proliferation marker Ki67 is negative here (H). At the margin of surgical resecate (MSR) is K14 expressed in all suprabasal layers (D), this aberrant expression is accompanied by loss of differentiation marker K10 (E) and high proliferation revealed by the presence of Ki67 (I). Similarly, epidermis overlaying dermal metastasis of melanoma is K14 positive even suprabasally (F); expression of K10 is dislocated more superficially (G). The epithelium in melanoma resecates reaches its thickest point on the periphery of the tumor margin (J). The tumor center is covered by the epithelium of variable thickness (J). Scale bars denote 25 μm.
Mentions: Significantly increased thickness of the epidermis in vicinity of tumor was observed in 90% of biopsies (more than 2 times compared to epidermal thickness at the surgical margin; mean 4.56×; median 3.84; range 0.72-12.2×, p value < 0.001 by Mann–Whitney test). The increased epidermal thickness was observed on the periphery of the tumor (Figure 1 J) without significant correlation to the value of Breslow’s depth of invasion.Figure 1

Bottom Line: Data were compared to results of transcriptome profiling of in vitro models, in which HPK were co-cultured with MC, normal human melanocytes, and NCSC, respectively.Differentially expressed candidate genes were verified by RT-qPCR.We conclude that the MC are able to influence locally the differentiation pattern of keratinocytes in vivo as well as in vitro.

View Article: PubMed Central - PubMed

Affiliation: 1st Faculty of Medicine, Institute of Anatomy, Charles University in Prague, U Nemocnice 3, CZ-12800 Prague, Czech Republic. strnad@img.cas.cz.

ABSTRACT

Background: Nodular melanoma is one of the most life threatening tumors with still poor therapeutic outcome. Similarly to other tumors, permissive microenvironment is essential for melanoma progression. Features of this microenvironment are arising from molecular crosstalk between the melanoma cells (MC) and the surrounding cell populations in the context of skin tissue. Here, we study the effect of melanoma cells on human primary keratinocytes (HPK). Presence of MC is as an important modulator of the tumor microenvironment and we compare it to the effect of nonmalignant lowly differentiated cells also originating from neural crest (NCSC).

Methods: Comparative morphometrical and immunohistochemical analysis of epidermis surrounding nodular melanoma (n = 100) was performed. Data were compared to results of transcriptome profiling of in vitro models, in which HPK were co-cultured with MC, normal human melanocytes, and NCSC, respectively. Differentially expressed candidate genes were verified by RT-qPCR. Biological activity of candidate proteins was assessed on cultured HPK.

Results: Epidermis surrounding nodular melanoma exhibits hyperplastic features in 90% of cases. This hyperplastic region exhibits aberrant suprabasal expression of keratin 14 accompanied by loss of keratin 10. We observe that MC and NCSC are able to increase expression of keratins 8, 14, 19, and vimentin in the co-cultured HPK. This in vitro finding partially correlates with pseudoepitheliomatous hyperplasia observed in melanoma biopsies. We provide evidence of FGF-2, CXCL-1, IL-8, and VEGF-A participation in the activity of melanoma cells on keratinocytes.

Conclusion: We conclude that the MC are able to influence locally the differentiation pattern of keratinocytes in vivo as well as in vitro. This interaction further highlights the role of intercellular interactions in melanoma. The reciprocal role of activated keratinocytes on biology of melanoma cells shall be verified in the future.

Show MeSH
Related in: MedlinePlus