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Characterization and identification of PARM-1 as a new potential oncogene.

Charfi C, Levros LC, Edouard E, Rassart E - Mol. Cancer (2013)

Bottom Line: Moreover, deletion mutants of human PARM-1 without either extracellular or cytoplasmic portions seem to retain the ability to induce anchorage-independent growth of NIH/3T3 cells.In addition, PARM-1 increases ERK1/2, but more importantly AKT and STAT3 phosphorylation.Our results strongly suggest the oncogenic potential of PARM-1.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratoire de Biologie Moléculaire, Département des Sciences Biologiques, Centre BioMed, Université du Québec à Montréal, Case Postale 8888, Succursale Centre-ville, Montréal, QC H3C-3P8, Canada.

ABSTRACT

Background: The Graffi murine retrovirus is a powerful tool to find leukemia associated oncogenes. Using DNA microarrays, we recently identified several genes specifically deregulated in T- and B-leukemias induced by this virus.

Results: In the present study, probsets associated with T-CD8+ leukemias were analyzed and we validated the expression profile of the Parm-1 gene. PARM-1 is a member of the mucin family. We showed that human PARM-1 is an intact secreted protein accumulating predominantly, such as murine PARM-1, at the Golgi and in the early and late endosomes. PARM-1 colocalization with α-tubulin suggests that its trafficking within the cell involves the microtubule cytoskeleton. Also, the protein co-localizes with caveolin-1 which probably mediates its internalization. Transient transfection of both mouse and human Parm-1 cDNAs conferred anchorage- and serum-independent growth and enhanced cell proliferation. Moreover, deletion mutants of human PARM-1 without either extracellular or cytoplasmic portions seem to retain the ability to induce anchorage-independent growth of NIH/3T3 cells. In addition, PARM-1 increases ERK1/2, but more importantly AKT and STAT3 phosphorylation.

Conclusions: Our results strongly suggest the oncogenic potential of PARM-1.

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Related in: MedlinePlus

Analysis of mParm-1 gene expression in sorted lymphoid leukemia samples. Semi-quantitative RT-PCR analysis of Parm-1 in 5 B and 5 T leukemias : (B4, Cd45+Cd19+Sca1+; B5, Cd45R+Cd19+Sca1+; B6, Cd45R+Cd19+Sca1+; B7, Cd45R+Cd19+Sca1+; B8, Cd45R+Cd19+Sca1+) (T4, Cd4+Cd8+; T5, Cd4+Cd8+; T6, Cd4+Cd8+; T7, Cd4+Cd8+; T8, Cd4+Cd8-). RT-PCRs were performed in triplicate using the following actin (forward primer: 5’- tgacggggtcacccacactgtgcccatcta-3’, reverse primer: 5’-ctagaagcatttgcggtggacgatggaggg-3’) and murine Parm-1 (forward primer: 5’- gttagctgttttggggacca-3’, reverse primer: 5’-cgtgcaaattagcatctgga-3’) specific primers. PCR products were analyzed on agarose gel and band density was quantified with Quantity One Image Software. The actin gene was used as internal control and expression levels in each leukemia are presented as a gene/actin density ratio. Data are representative of 3 independent experiments. Statistical analysis was performed using one-way analysis of variance, and P lower to 0.05 was considered to be significant (*P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001) compared with their respective control (CB2, B cells from normal spleen and CT2, T cells from normal thymus).
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Figure 1: Analysis of mParm-1 gene expression in sorted lymphoid leukemia samples. Semi-quantitative RT-PCR analysis of Parm-1 in 5 B and 5 T leukemias : (B4, Cd45+Cd19+Sca1+; B5, Cd45R+Cd19+Sca1+; B6, Cd45R+Cd19+Sca1+; B7, Cd45R+Cd19+Sca1+; B8, Cd45R+Cd19+Sca1+) (T4, Cd4+Cd8+; T5, Cd4+Cd8+; T6, Cd4+Cd8+; T7, Cd4+Cd8+; T8, Cd4+Cd8-). RT-PCRs were performed in triplicate using the following actin (forward primer: 5’- tgacggggtcacccacactgtgcccatcta-3’, reverse primer: 5’-ctagaagcatttgcggtggacgatggaggg-3’) and murine Parm-1 (forward primer: 5’- gttagctgttttggggacca-3’, reverse primer: 5’-cgtgcaaattagcatctgga-3’) specific primers. PCR products were analyzed on agarose gel and band density was quantified with Quantity One Image Software. The actin gene was used as internal control and expression levels in each leukemia are presented as a gene/actin density ratio. Data are representative of 3 independent experiments. Statistical analysis was performed using one-way analysis of variance, and P lower to 0.05 was considered to be significant (*P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001) compared with their respective control (CB2, B cells from normal spleen and CT2, T cells from normal thymus).

Mentions: We focused on the mParm-1 (9130213B05Rik) gene. The expression level of mParm-1 was measured by semi-quantitative RT-PCR in several Graffi MuLV-induced tumors. Significant over-expression was only observed in T-CD8+ tumors when compared to control T-cells. This result confirms the specificity of the mParm-1 gene up-regulation to T-CD8+ leukemias (Figure 1).


Characterization and identification of PARM-1 as a new potential oncogene.

Charfi C, Levros LC, Edouard E, Rassart E - Mol. Cancer (2013)

Analysis of mParm-1 gene expression in sorted lymphoid leukemia samples. Semi-quantitative RT-PCR analysis of Parm-1 in 5 B and 5 T leukemias : (B4, Cd45+Cd19+Sca1+; B5, Cd45R+Cd19+Sca1+; B6, Cd45R+Cd19+Sca1+; B7, Cd45R+Cd19+Sca1+; B8, Cd45R+Cd19+Sca1+) (T4, Cd4+Cd8+; T5, Cd4+Cd8+; T6, Cd4+Cd8+; T7, Cd4+Cd8+; T8, Cd4+Cd8-). RT-PCRs were performed in triplicate using the following actin (forward primer: 5’- tgacggggtcacccacactgtgcccatcta-3’, reverse primer: 5’-ctagaagcatttgcggtggacgatggaggg-3’) and murine Parm-1 (forward primer: 5’- gttagctgttttggggacca-3’, reverse primer: 5’-cgtgcaaattagcatctgga-3’) specific primers. PCR products were analyzed on agarose gel and band density was quantified with Quantity One Image Software. The actin gene was used as internal control and expression levels in each leukemia are presented as a gene/actin density ratio. Data are representative of 3 independent experiments. Statistical analysis was performed using one-way analysis of variance, and P lower to 0.05 was considered to be significant (*P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001) compared with their respective control (CB2, B cells from normal spleen and CT2, T cells from normal thymus).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750824&req=5

Figure 1: Analysis of mParm-1 gene expression in sorted lymphoid leukemia samples. Semi-quantitative RT-PCR analysis of Parm-1 in 5 B and 5 T leukemias : (B4, Cd45+Cd19+Sca1+; B5, Cd45R+Cd19+Sca1+; B6, Cd45R+Cd19+Sca1+; B7, Cd45R+Cd19+Sca1+; B8, Cd45R+Cd19+Sca1+) (T4, Cd4+Cd8+; T5, Cd4+Cd8+; T6, Cd4+Cd8+; T7, Cd4+Cd8+; T8, Cd4+Cd8-). RT-PCRs were performed in triplicate using the following actin (forward primer: 5’- tgacggggtcacccacactgtgcccatcta-3’, reverse primer: 5’-ctagaagcatttgcggtggacgatggaggg-3’) and murine Parm-1 (forward primer: 5’- gttagctgttttggggacca-3’, reverse primer: 5’-cgtgcaaattagcatctgga-3’) specific primers. PCR products were analyzed on agarose gel and band density was quantified with Quantity One Image Software. The actin gene was used as internal control and expression levels in each leukemia are presented as a gene/actin density ratio. Data are representative of 3 independent experiments. Statistical analysis was performed using one-way analysis of variance, and P lower to 0.05 was considered to be significant (*P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001) compared with their respective control (CB2, B cells from normal spleen and CT2, T cells from normal thymus).
Mentions: We focused on the mParm-1 (9130213B05Rik) gene. The expression level of mParm-1 was measured by semi-quantitative RT-PCR in several Graffi MuLV-induced tumors. Significant over-expression was only observed in T-CD8+ tumors when compared to control T-cells. This result confirms the specificity of the mParm-1 gene up-regulation to T-CD8+ leukemias (Figure 1).

Bottom Line: Moreover, deletion mutants of human PARM-1 without either extracellular or cytoplasmic portions seem to retain the ability to induce anchorage-independent growth of NIH/3T3 cells.In addition, PARM-1 increases ERK1/2, but more importantly AKT and STAT3 phosphorylation.Our results strongly suggest the oncogenic potential of PARM-1.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratoire de Biologie Moléculaire, Département des Sciences Biologiques, Centre BioMed, Université du Québec à Montréal, Case Postale 8888, Succursale Centre-ville, Montréal, QC H3C-3P8, Canada.

ABSTRACT

Background: The Graffi murine retrovirus is a powerful tool to find leukemia associated oncogenes. Using DNA microarrays, we recently identified several genes specifically deregulated in T- and B-leukemias induced by this virus.

Results: In the present study, probsets associated with T-CD8+ leukemias were analyzed and we validated the expression profile of the Parm-1 gene. PARM-1 is a member of the mucin family. We showed that human PARM-1 is an intact secreted protein accumulating predominantly, such as murine PARM-1, at the Golgi and in the early and late endosomes. PARM-1 colocalization with α-tubulin suggests that its trafficking within the cell involves the microtubule cytoskeleton. Also, the protein co-localizes with caveolin-1 which probably mediates its internalization. Transient transfection of both mouse and human Parm-1 cDNAs conferred anchorage- and serum-independent growth and enhanced cell proliferation. Moreover, deletion mutants of human PARM-1 without either extracellular or cytoplasmic portions seem to retain the ability to induce anchorage-independent growth of NIH/3T3 cells. In addition, PARM-1 increases ERK1/2, but more importantly AKT and STAT3 phosphorylation.

Conclusions: Our results strongly suggest the oncogenic potential of PARM-1.

Show MeSH
Related in: MedlinePlus