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Knock-down of PRAME increases retinoic acid signaling and cytotoxic drug sensitivity of Hodgkin lymphoma cells.

Kewitz S, Staege MS - PLoS ONE (2013)

Bottom Line: DNA microarray analysis of HL cells after PRAME knock-down indicated regulation of several genes including down-regulation of known anti-apoptotic factors.Increased retinoic acid signaling in these cells was revealed by increased expression of the retinoic acid metabolizing cytochrome P450 (CYP26B1), a transcriptional target of retinoic acid signaling.Our data suggest that PRAME inhibits retinoic acid signaling in HL cells and that the knock-down of PRAME might be an interesting option for the development of new therapy strategies for patients with chemo-resistant HL.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Martin-Luther-University Halle-Wittenberg, Halle, Germany.

ABSTRACT
The prognosis for patients with Hodgkin lymphoma (HL) has improved in recent decades. On the other hand, not all patients can be cured with the currently established therapy regimes and this therapy is associated with several adverse late effects. Therefore it is necessary to develop new therapy strategies. After treatment of L-540 HL cells with 5'-azacytidine (5AC), we observed increased expression of the preferentially expressed antigen in melanoma (PRAME). In addition, we detected an increased resistance of 5AC-treated cells against cytotoxic drugs. We analyzed the influence of PRAME on cell survival of HL cells by knocking down PRAME in the chemotherapy resistant cell line L-428, a cell line that express PRAME at a high level. After knock-down of PRAME using vector based RNA interference we observed increased sensitivity for cisplatin, etoposide and retinoic acid. DNA microarray analysis of HL cells after PRAME knock-down indicated regulation of several genes including down-regulation of known anti-apoptotic factors. Increased retinoic acid signaling in these cells was revealed by increased expression of the retinoic acid metabolizing cytochrome P450 (CYP26B1), a transcriptional target of retinoic acid signaling. Our data suggest that PRAME inhibits retinoic acid signaling in HL cells and that the knock-down of PRAME might be an interesting option for the development of new therapy strategies for patients with chemo-resistant HL.

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Increased induction of CYP26B1 in L-428 cells after knock-down of PRAME.Expression of CYP26B1 was analyzed in HL cell line L-428 with and without knock-down of PRAME by qRT-PCR. Cells were treated with ATRA or DMSO for four days. Presented are means and standard errors from six/eleven determinations. For comparative analysis, the means of cells on day zero was set as 1. Asterisks indicate significance (p<0.05; Students t test).
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pone-0055897-g004: Increased induction of CYP26B1 in L-428 cells after knock-down of PRAME.Expression of CYP26B1 was analyzed in HL cell line L-428 with and without knock-down of PRAME by qRT-PCR. Cells were treated with ATRA or DMSO for four days. Presented are means and standard errors from six/eleven determinations. For comparative analysis, the means of cells on day zero was set as 1. Asterisks indicate significance (p<0.05; Students t test).

Mentions: These data suggested that knock-down of PRAME increased the sensitivity for retinoic acid signaling. We analyzed the influence of PRAME on expression of RA target genes. We treated cells with knock-down of PRAME and cells with the control vector with 2.5×10−4 M all-trans RA. After four days we harvested the cells, isolated RNA and analyzed expression of the cytochrome P450 family member 26B1 (CYP26B1). CYP26B1 is involved in the metabolism of retinoic acid and a target of retinoic acid signaling [28]. As shown in Figure 4, expression of CYP26B1 increased after RA treatment. Control cells incubated with all-trans RA expressed 63.7 times higher levels of CYP26B1 than untreated cells. Cells with PRAME knock-down expressed 222 times higher levels, indicating that suppression of PRAME resulted in increased RA signaling.


Knock-down of PRAME increases retinoic acid signaling and cytotoxic drug sensitivity of Hodgkin lymphoma cells.

Kewitz S, Staege MS - PLoS ONE (2013)

Increased induction of CYP26B1 in L-428 cells after knock-down of PRAME.Expression of CYP26B1 was analyzed in HL cell line L-428 with and without knock-down of PRAME by qRT-PCR. Cells were treated with ATRA or DMSO for four days. Presented are means and standard errors from six/eleven determinations. For comparative analysis, the means of cells on day zero was set as 1. Asterisks indicate significance (p<0.05; Students t test).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3569423&req=5

pone-0055897-g004: Increased induction of CYP26B1 in L-428 cells after knock-down of PRAME.Expression of CYP26B1 was analyzed in HL cell line L-428 with and without knock-down of PRAME by qRT-PCR. Cells were treated with ATRA or DMSO for four days. Presented are means and standard errors from six/eleven determinations. For comparative analysis, the means of cells on day zero was set as 1. Asterisks indicate significance (p<0.05; Students t test).
Mentions: These data suggested that knock-down of PRAME increased the sensitivity for retinoic acid signaling. We analyzed the influence of PRAME on expression of RA target genes. We treated cells with knock-down of PRAME and cells with the control vector with 2.5×10−4 M all-trans RA. After four days we harvested the cells, isolated RNA and analyzed expression of the cytochrome P450 family member 26B1 (CYP26B1). CYP26B1 is involved in the metabolism of retinoic acid and a target of retinoic acid signaling [28]. As shown in Figure 4, expression of CYP26B1 increased after RA treatment. Control cells incubated with all-trans RA expressed 63.7 times higher levels of CYP26B1 than untreated cells. Cells with PRAME knock-down expressed 222 times higher levels, indicating that suppression of PRAME resulted in increased RA signaling.

Bottom Line: DNA microarray analysis of HL cells after PRAME knock-down indicated regulation of several genes including down-regulation of known anti-apoptotic factors.Increased retinoic acid signaling in these cells was revealed by increased expression of the retinoic acid metabolizing cytochrome P450 (CYP26B1), a transcriptional target of retinoic acid signaling.Our data suggest that PRAME inhibits retinoic acid signaling in HL cells and that the knock-down of PRAME might be an interesting option for the development of new therapy strategies for patients with chemo-resistant HL.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Martin-Luther-University Halle-Wittenberg, Halle, Germany.

ABSTRACT
The prognosis for patients with Hodgkin lymphoma (HL) has improved in recent decades. On the other hand, not all patients can be cured with the currently established therapy regimes and this therapy is associated with several adverse late effects. Therefore it is necessary to develop new therapy strategies. After treatment of L-540 HL cells with 5'-azacytidine (5AC), we observed increased expression of the preferentially expressed antigen in melanoma (PRAME). In addition, we detected an increased resistance of 5AC-treated cells against cytotoxic drugs. We analyzed the influence of PRAME on cell survival of HL cells by knocking down PRAME in the chemotherapy resistant cell line L-428, a cell line that express PRAME at a high level. After knock-down of PRAME using vector based RNA interference we observed increased sensitivity for cisplatin, etoposide and retinoic acid. DNA microarray analysis of HL cells after PRAME knock-down indicated regulation of several genes including down-regulation of known anti-apoptotic factors. Increased retinoic acid signaling in these cells was revealed by increased expression of the retinoic acid metabolizing cytochrome P450 (CYP26B1), a transcriptional target of retinoic acid signaling. Our data suggest that PRAME inhibits retinoic acid signaling in HL cells and that the knock-down of PRAME might be an interesting option for the development of new therapy strategies for patients with chemo-resistant HL.

Show MeSH
Related in: MedlinePlus