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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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Anchorage-independent growth. The effect of Msi1 expression upon anchorage-independent growth was studied as an in vitro indicator of tumorigenicity in Daoy cells. Two independent experiments were performed with similar results. Msi1 depletion reduced the ability to form colonies in soft agar by 3 to 4-fold which indicated that Msi1 may sustain the malignant potential of this tumor cell line. a) A representative image of the colonies formed by the control and knockdown (KD) cell lines is shown. The average results for the three clones (b) and for each individual one (c) are represented. * and ** indicate p < 10-3 and p < 10-5, respectively, compared to control.
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Figure 2: Anchorage-independent growth. The effect of Msi1 expression upon anchorage-independent growth was studied as an in vitro indicator of tumorigenicity in Daoy cells. Two independent experiments were performed with similar results. Msi1 depletion reduced the ability to form colonies in soft agar by 3 to 4-fold which indicated that Msi1 may sustain the malignant potential of this tumor cell line. a) A representative image of the colonies formed by the control and knockdown (KD) cell lines is shown. The average results for the three clones (b) and for each individual one (c) are represented. * and ** indicate p < 10-3 and p < 10-5, respectively, compared to control.

Mentions: We analyzed the effect of Msi1 on anchorage-independent growth (clonogenicity) in Daoy cells as an in vitro assay for tumorigenicity. Colony formation assays were performed in soft agar as previously described [21]. As shown in Figure 2, Msi1 knockdown reduced the ability of Daoy cells to form colonies in soft agar by 3 to 4-fold thus indicating that Msi1 promoted cell proliferation in these cancer cells and, consequently, that might play a role in tumor growth.


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)

Anchorage-independent growth. The effect of Msi1 expression upon anchorage-independent growth was studied as an in vitro indicator of tumorigenicity in Daoy cells. Two independent experiments were performed with similar results. Msi1 depletion reduced the ability to form colonies in soft agar by 3 to 4-fold which indicated that Msi1 may sustain the malignant potential of this tumor cell line. a) A representative image of the colonies formed by the control and knockdown (KD) cell lines is shown. The average results for the three clones (b) and for each individual one (c) are represented. * and ** indicate p < 10-3 and p < 10-5, respectively, compared to control.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 2: Anchorage-independent growth. The effect of Msi1 expression upon anchorage-independent growth was studied as an in vitro indicator of tumorigenicity in Daoy cells. Two independent experiments were performed with similar results. Msi1 depletion reduced the ability to form colonies in soft agar by 3 to 4-fold which indicated that Msi1 may sustain the malignant potential of this tumor cell line. a) A representative image of the colonies formed by the control and knockdown (KD) cell lines is shown. The average results for the three clones (b) and for each individual one (c) are represented. * and ** indicate p < 10-3 and p < 10-5, respectively, compared to control.
Mentions: We analyzed the effect of Msi1 on anchorage-independent growth (clonogenicity) in Daoy cells as an in vitro assay for tumorigenicity. Colony formation assays were performed in soft agar as previously described [21]. As shown in Figure 2, Msi1 knockdown reduced the ability of Daoy cells to form colonies in soft agar by 3 to 4-fold thus indicating that Msi1 promoted cell proliferation in these cancer cells and, consequently, that might play a role in tumor growth.

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