Abstract
We have previously identified 1 241 regions of somatic copy number alterations (CNAs) in hepatocellular carcinoma (HCC). In the present study, we found that a novel recurrent focal amplicon, 1q24.1-24.2, targets the MPZL1 gene in HCC. Notably, there is a positive correlation between the expression levels of MPZL1 and intrahepatic metastasis of the HCC specimens. MPZL1 can significantly enhance the migratory and metastatic potential of the HCC cells. Moreover, we found that one of the mechanisms by which MPZL1 promotes HCC cell migration is by inducing the phosphorylation and activation of the pro-metastatic protein, cortactin. Additionally, we found that Src kinase mediates the phosphorylation and activation of cortactin induced by MPZL1 overexpression. Taken together, these findings suggest that MPZL1 is a novel pro-metastatic gene targeted by a recurrent region of copy number amplification at 1q24.1-24.2 in HCC.
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Acknowledgements
We are very grateful to professor Didier Trono (Ecole Polytechnique Fédérale de Lausanne, 1015 Lausanne, Switzerland) for providing the pWPXL, psPAX2 and pMD2.G lentivirus plasmids . This work was partially supported by the National Key Basic Research Program of China (2013CB910500), the National Natural Science Foundation of China (81125016, 81071637 and 91029728), Shanghai Science and Technology Commission (11XD1404500), Shanghai Municipal Education Commission and Shanghai Municipal Health Bureau (11SG18 and XBR2011039).
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( Supplementary information is linked to the online version of the paper on the Cell Research website.)
Supplementary information
Supplementary information, Table S1
Differentially expressed genes within the broad region of 1q24 copy number gain (PDF 77 kb)
Supplementary information, Table S2
Correlation of MPZL1 expression with multiple clinical features of HCC specimens. (PDF 1247 kb)
Supplementary information, Table S3
Summary of results derived from phospho-antibody array (PDF 46 kb)
Supplementary information, Table S4
Proteins for which the phosphorylation states increased in HUH-7 cells overexpressing MPZL1 (PDF 59 kb)
Supplementary information, Table S5
The sequences of siRNAs and shRNA plasmids used (PDF 33 kb)
Supplementary information, Table S6
Primers used for q-PCR and subclone (PDF 91 kb)
Supplementary information, Table S7
Clinical and pathological features of 58 HCC specimens (PDF 58 kb)
Supplementary information, Figure S1
Examination of the DNA dosages and expression levels of four candidate genes in HCCs. (PDF 173 kb)
Supplementary information, Figure S2
Examination of the relative expression levels of MPZL1 isoforms in HCCs. (PDF 141 kb)
Supplementary information, Figure S3
The expression levels of MPZL1 in HCC derived from gene expression arrays. (PDF 130 kb)
Supplementary information, Figure S4
Knockdown of the MPZL1 gene inhibited HCC cell migration, but not cell proliferation. (PDF 231 kb)
Supplementary information, Figure S5
Enforced expression of MPZL1 has no significant effect on HCC cell proliferation. (PDF 75 kb)
Supplementary information, Figure S6
Knockdown of the CTTN gene inhibits the migratory and invasive abilities of HCC cells. (PDF 148 kb)
Supplementary information, Figure S7
Ectopic expression of MPZL1 leads to decreased phosphorylation levels of negative regulatory site of Src (Y527). (PDF 58 kb)
Supplementary information, Figure S8
The Detection and quantitation of MPZL1 methylation in 37 paired HCC and adjacent paracancer tissues by real-time methylation-specific PCR. (PDF 64 kb)
Supplementary information, Figure S9
The overexpression of MPZL1 in HUH-7 cells induces the phosphorylation of the Erk-1/2 (T202/Y204) and AKT (S473) proteins. (PDF 151 kb)
Supplementary information, Figure S10
The detection of siRNA mediated knockdown of targeted genes (CTTN, SHP-2 and Src) in HCC cells by immunoblotting. (PDF 189 kb)
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Jia, D., Jing, Y., Zhang, Z. et al. Amplification of MPZL1/PZR promotes tumor cell migration through Src-mediated phosphorylation of cortactin in hepatocellular carcinoma. Cell Res 24, 204–217 (2014). https://doi.org/10.1038/cr.2013.158
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DOI: https://doi.org/10.1038/cr.2013.158
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