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Musashi1 modulates cell proliferation genes in the medulloblastoma cell line Daoy.

Sanchez-Diaz PC, Burton TL, Burns SC, Hung JY, Penalva LO - BMC Cancer (2008)

Bottom Line: We used the human medulloblastoma cell line Daoy as model system in this study to knock down the expression of Msi1 and determine the effects upon soft agar growth and neurophere formation.We observed that MSI1 expression was elevated in Daoy cells cultured as neurospheres compared to those grown as monolayer.Moreover, differential expression of a group of Notch, Hedgehog and Wnt pathway related genes including MYCN, FOS, NOTCH2, SMO, CDKN1A, CCND2, CCND1, and DKK1, was also found in the Msi1 knockdown, demonstrating that Msi1 modulated the expression of a subset of cell proliferation, differentiation and survival genes in Daoy.

View Article: PubMed Central - HTML - PubMed

Affiliation: Greehey Children's Cancer Research Institute, University of Texas Health Science Center at San Antonio, TX, USA. sanchezdiaz@uthscsa.edu

ABSTRACT

Background: Musashi1 (Msi1) is an RNA binding protein with a central role during nervous system development and stem cell maintenance. High levels of Msi1 have been reported in several malignancies including brain tumors thereby associating Msi1 and cancer.

Methods: We used the human medulloblastoma cell line Daoy as model system in this study to knock down the expression of Msi1 and determine the effects upon soft agar growth and neurophere formation. Quantitative RT-PCR was conducted to evaluate the expression of cell proliferation, differentiation and survival genes in Msi1 depleted Daoy cells.

Results: We observed that MSI1 expression was elevated in Daoy cells cultured as neurospheres compared to those grown as monolayer. These data indicated that Msi1 might be involved in regulating proliferation in cancer cells. Here we show that shRNA mediated Msi1 depletion in Daoy cells notably impaired their ability to form colonies in soft agar and to grow as neurospheres in culture. Moreover, differential expression of a group of Notch, Hedgehog and Wnt pathway related genes including MYCN, FOS, NOTCH2, SMO, CDKN1A, CCND2, CCND1, and DKK1, was also found in the Msi1 knockdown, demonstrating that Msi1 modulated the expression of a subset of cell proliferation, differentiation and survival genes in Daoy.

Conclusion: Our data suggested that Msi1 may promote cancer cell proliferation and survival as its loss seems to have a detrimental effect in the maintenance of medulloblastoma cancer cells. In this regard, Msi1 might be a positive regulator of tumor progression and a potential target for therapy.

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Msi1 expression in Daoy neurospheres. The levels of MSI1 RNA levels were analyzed in Daoy cells grown as monolayer (nonNSC) and as neurosphere (NSC) cultures using quantitative RT-PCR. GAPDH and 15S mRNA levels were used as endogenous control for normalization. On average, a 2-fold increase in MSI1 was detected in the neurosphere cultures.* indicate p < 0.01.
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Figure 1: Msi1 expression in Daoy neurospheres. The levels of MSI1 RNA levels were analyzed in Daoy cells grown as monolayer (nonNSC) and as neurosphere (NSC) cultures using quantitative RT-PCR. GAPDH and 15S mRNA levels were used as endogenous control for normalization. On average, a 2-fold increase in MSI1 was detected in the neurosphere cultures.* indicate p < 0.01.

Mentions: To determine if Msi1 might play a role in Daoy cancer cell proliferation, we measured the levels of MSI1 mRNA in Daoy neurosphere cultures by quantitative RT-PCR (Figure 1). On average, a 2-fold increase in MSI1 level was detected in neurospheres (actively proliferating culture) compared to monolayers (less actively proliferating culture). Since neurosphere cultures can be enriched in tumor re-initiating cells and Msi1 is a stem cell marker with a role in cell cycle progression [13], the higher level of Msi1 detected in neurospheres indicated that Msi1 may contribute to Daoy cancer cell proliferation.


Musashi1 modulates cell proliferation genes in the medulloblastoma cell line Daoy.

Sanchez-Diaz PC, Burton TL, Burns SC, Hung JY, Penalva LO - BMC Cancer (2008)

Msi1 expression in Daoy neurospheres. The levels of MSI1 RNA levels were analyzed in Daoy cells grown as monolayer (nonNSC) and as neurosphere (NSC) cultures using quantitative RT-PCR. GAPDH and 15S mRNA levels were used as endogenous control for normalization. On average, a 2-fold increase in MSI1 was detected in the neurosphere cultures.* indicate p < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2572071&req=5

Figure 1: Msi1 expression in Daoy neurospheres. The levels of MSI1 RNA levels were analyzed in Daoy cells grown as monolayer (nonNSC) and as neurosphere (NSC) cultures using quantitative RT-PCR. GAPDH and 15S mRNA levels were used as endogenous control for normalization. On average, a 2-fold increase in MSI1 was detected in the neurosphere cultures.* indicate p < 0.01.
Mentions: To determine if Msi1 might play a role in Daoy cancer cell proliferation, we measured the levels of MSI1 mRNA in Daoy neurosphere cultures by quantitative RT-PCR (Figure 1). On average, a 2-fold increase in MSI1 level was detected in neurospheres (actively proliferating culture) compared to monolayers (less actively proliferating culture). Since neurosphere cultures can be enriched in tumor re-initiating cells and Msi1 is a stem cell marker with a role in cell cycle progression [13], the higher level of Msi1 detected in neurospheres indicated that Msi1 may contribute to Daoy cancer cell proliferation.

Bottom Line: We used the human medulloblastoma cell line Daoy as model system in this study to knock down the expression of Msi1 and determine the effects upon soft agar growth and neurophere formation.We observed that MSI1 expression was elevated in Daoy cells cultured as neurospheres compared to those grown as monolayer.Moreover, differential expression of a group of Notch, Hedgehog and Wnt pathway related genes including MYCN, FOS, NOTCH2, SMO, CDKN1A, CCND2, CCND1, and DKK1, was also found in the Msi1 knockdown, demonstrating that Msi1 modulated the expression of a subset of cell proliferation, differentiation and survival genes in Daoy.

View Article: PubMed Central - HTML - PubMed

Affiliation: Greehey Children's Cancer Research Institute, University of Texas Health Science Center at San Antonio, TX, USA. sanchezdiaz@uthscsa.edu

ABSTRACT

Background: Musashi1 (Msi1) is an RNA binding protein with a central role during nervous system development and stem cell maintenance. High levels of Msi1 have been reported in several malignancies including brain tumors thereby associating Msi1 and cancer.

Methods: We used the human medulloblastoma cell line Daoy as model system in this study to knock down the expression of Msi1 and determine the effects upon soft agar growth and neurophere formation. Quantitative RT-PCR was conducted to evaluate the expression of cell proliferation, differentiation and survival genes in Msi1 depleted Daoy cells.

Results: We observed that MSI1 expression was elevated in Daoy cells cultured as neurospheres compared to those grown as monolayer. These data indicated that Msi1 might be involved in regulating proliferation in cancer cells. Here we show that shRNA mediated Msi1 depletion in Daoy cells notably impaired their ability to form colonies in soft agar and to grow as neurospheres in culture. Moreover, differential expression of a group of Notch, Hedgehog and Wnt pathway related genes including MYCN, FOS, NOTCH2, SMO, CDKN1A, CCND2, CCND1, and DKK1, was also found in the Msi1 knockdown, demonstrating that Msi1 modulated the expression of a subset of cell proliferation, differentiation and survival genes in Daoy.

Conclusion: Our data suggested that Msi1 may promote cancer cell proliferation and survival as its loss seems to have a detrimental effect in the maintenance of medulloblastoma cancer cells. In this regard, Msi1 might be a positive regulator of tumor progression and a potential target for therapy.

Show MeSH
Related in: MedlinePlus