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EMT-associated factors promote invasive properties of uveal melanoma cells.

Asnaghi L, Gezgin G, Tripathy A, Handa JT, Merbs SL, van der Velden PA, Jager MJ, Harbour JW, Eberhart CG - Mol. Vis. (2015)

Bottom Line: The genetic downregulation of ZEB1 in OCM1, OMM1, and 92.1 resulted in a more than 50% reduction in invasion, but only suppressed growth in OMM1 cells.Suppression of Twist1 in Mel290 and OMM1 reduced growth and invasion by more than 50%.The downregulation of Snail1 in the 92.1 cell line reduced invasion by 50%, but did not interfere with growth.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Johns Hopkins University, School of Medicine, Baltimore, MD.

ABSTRACT

Purpose: Transcription factors regulating the epithelial-to-mesenchymal transition (EMT) program contribute to carcinogenesis and metastasis in many tumors, including cutaneous melanoma. However, little is known about the role of EMT factors in the growth and metastatic dissemination of uveal melanoma cells. Here, we analyzed the expression and functions of the EMT factors ZEB1, Twist1, and Snail1 in uveal melanoma cell lines and primary tumors.

Methods: ZEB1, Twist1, and Snail1 mRNA levels were measured using qPCR in five uveal melanoma cell lines and in 30 primary tumors. Gene expression was used to determine class 1 and class 2 signatures in the primary tumors. Short hairpin RNA was used to downregulate the expressions of the EMT factors; then, growth and transwell invasion assays were performed.

Results: ZEB1, Twist1, and Snail1 were expressed in all five uveal melanoma lines, with ZEB1 having the highest protein levels. ZEB1 mRNA was significantly elevated in highly metastatic class 2 primary tumors for which survival data were not available, whereas a high gene expression of Twist1 was associated with a worse prognosis in a separate tumor cohort analyzed by expression profiling. The genetic downregulation of ZEB1 in OCM1, OMM1, and 92.1 resulted in a more than 50% reduction in invasion, but only suppressed growth in OMM1 cells. Suppression of Twist1 in Mel290 and OMM1 reduced growth and invasion by more than 50%. The downregulation of Snail1 in the 92.1 cell line reduced invasion by 50%, but did not interfere with growth.

Conclusions: The downregulation of ZEB1, Twist1, and Snail1 reduces the invasive properties of uveal melanoma cells, and the elevated mRNA levels of ZEB1 and Twist1 are associated with a more aggressive clinical phenotype in uveal melanoma samples. Therefore, these factors could represent new therapeutic targets in patients with ocular melanoma.

No MeSH data available.


Related in: MedlinePlus

Increased ZEB1, Twist1, and Snail1 mRNA levels in invasive uveal melanoma cells. The mRNA levels of the EMT-factors ZEB1, Twist1, and Snail1 were significantly increased, as measured by qPCR, in invading OMM1 cells, which moved through a Matrigel-coated filter, as compared to non-invading OMM1 cells. Data shown are mean ± standard deviation (SD); *p = 0.028; **p = 0.002; ***p = 0.0005 versus non-invading cells.
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f2: Increased ZEB1, Twist1, and Snail1 mRNA levels in invasive uveal melanoma cells. The mRNA levels of the EMT-factors ZEB1, Twist1, and Snail1 were significantly increased, as measured by qPCR, in invading OMM1 cells, which moved through a Matrigel-coated filter, as compared to non-invading OMM1 cells. Data shown are mean ± standard deviation (SD); *p = 0.028; **p = 0.002; ***p = 0.0005 versus non-invading cells.

Mentions: To determine whether ZEB1, Twist1, and Snail1 increase migration and promote the metastatic spread of uveal melanoma, we determined the mRNA levels of these three transcription factors in OMM1 cells that had moved overnight through a microporous membrane precoated with Matrigel, which mimics the composition and the properties of the ECM (Figure 2). We observed that all these three factors were significantly increased at the transcriptional level in the invasive cells harvested from the lower surface of the Matrigel-coated membrane, as compared to those that did not invade and were still located in the upper part of the filter after overnight incubation.


EMT-associated factors promote invasive properties of uveal melanoma cells.

Asnaghi L, Gezgin G, Tripathy A, Handa JT, Merbs SL, van der Velden PA, Jager MJ, Harbour JW, Eberhart CG - Mol. Vis. (2015)

Increased ZEB1, Twist1, and Snail1 mRNA levels in invasive uveal melanoma cells. The mRNA levels of the EMT-factors ZEB1, Twist1, and Snail1 were significantly increased, as measured by qPCR, in invading OMM1 cells, which moved through a Matrigel-coated filter, as compared to non-invading OMM1 cells. Data shown are mean ± standard deviation (SD); *p = 0.028; **p = 0.002; ***p = 0.0005 versus non-invading cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4548792&req=5

f2: Increased ZEB1, Twist1, and Snail1 mRNA levels in invasive uveal melanoma cells. The mRNA levels of the EMT-factors ZEB1, Twist1, and Snail1 were significantly increased, as measured by qPCR, in invading OMM1 cells, which moved through a Matrigel-coated filter, as compared to non-invading OMM1 cells. Data shown are mean ± standard deviation (SD); *p = 0.028; **p = 0.002; ***p = 0.0005 versus non-invading cells.
Mentions: To determine whether ZEB1, Twist1, and Snail1 increase migration and promote the metastatic spread of uveal melanoma, we determined the mRNA levels of these three transcription factors in OMM1 cells that had moved overnight through a microporous membrane precoated with Matrigel, which mimics the composition and the properties of the ECM (Figure 2). We observed that all these three factors were significantly increased at the transcriptional level in the invasive cells harvested from the lower surface of the Matrigel-coated membrane, as compared to those that did not invade and were still located in the upper part of the filter after overnight incubation.

Bottom Line: The genetic downregulation of ZEB1 in OCM1, OMM1, and 92.1 resulted in a more than 50% reduction in invasion, but only suppressed growth in OMM1 cells.Suppression of Twist1 in Mel290 and OMM1 reduced growth and invasion by more than 50%.The downregulation of Snail1 in the 92.1 cell line reduced invasion by 50%, but did not interfere with growth.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Johns Hopkins University, School of Medicine, Baltimore, MD.

ABSTRACT

Purpose: Transcription factors regulating the epithelial-to-mesenchymal transition (EMT) program contribute to carcinogenesis and metastasis in many tumors, including cutaneous melanoma. However, little is known about the role of EMT factors in the growth and metastatic dissemination of uveal melanoma cells. Here, we analyzed the expression and functions of the EMT factors ZEB1, Twist1, and Snail1 in uveal melanoma cell lines and primary tumors.

Methods: ZEB1, Twist1, and Snail1 mRNA levels were measured using qPCR in five uveal melanoma cell lines and in 30 primary tumors. Gene expression was used to determine class 1 and class 2 signatures in the primary tumors. Short hairpin RNA was used to downregulate the expressions of the EMT factors; then, growth and transwell invasion assays were performed.

Results: ZEB1, Twist1, and Snail1 were expressed in all five uveal melanoma lines, with ZEB1 having the highest protein levels. ZEB1 mRNA was significantly elevated in highly metastatic class 2 primary tumors for which survival data were not available, whereas a high gene expression of Twist1 was associated with a worse prognosis in a separate tumor cohort analyzed by expression profiling. The genetic downregulation of ZEB1 in OCM1, OMM1, and 92.1 resulted in a more than 50% reduction in invasion, but only suppressed growth in OMM1 cells. Suppression of Twist1 in Mel290 and OMM1 reduced growth and invasion by more than 50%. The downregulation of Snail1 in the 92.1 cell line reduced invasion by 50%, but did not interfere with growth.

Conclusions: The downregulation of ZEB1, Twist1, and Snail1 reduces the invasive properties of uveal melanoma cells, and the elevated mRNA levels of ZEB1 and Twist1 are associated with a more aggressive clinical phenotype in uveal melanoma samples. Therefore, these factors could represent new therapeutic targets in patients with ocular melanoma.

No MeSH data available.


Related in: MedlinePlus