Limits...
DDX5 promotes proliferation and tumorigenesis of non-small-cell lung cancer cells by activating β-catenin signaling pathway.

Wang Z, Luo Z, Zhou L, Li X, Jiang T, Fu E - Cancer Sci. (2015)

Bottom Line: The DEAD-box-protein DDX5 is an ATP-dependent RNA helicase that is frequently overexpressed in various cancers and acts as a transcriptional co-activator of several transcription factors, including β-catenin.DDX5 is reported to be involved in cancer progression by promoting cell proliferation and epithelial-mesenchymal transition.We found that DDX5 was significantly overexpressed in NSCLC tissues as compared with the matched normal adjacent tissues.

View Article: PubMed Central - PubMed

Affiliation: Department of Thoracic Surgery, Tangdu Hospital, The Fourth Military Medical University, Xi'an, China.

No MeSH data available.


Related in: MedlinePlus

DDX5 promotes β-catenin nuclear accumulation and activates cyclin D1 and c-Myc transcription. (a) Co-immunoprecipitation (IP) analysis of DDX5 and β-catenin in H520 and A549 non-small-cell lung cancer (NSCLC) cells. β-catenin and DDX5 can be reciprocally co-immunoprecipitated by their antibodies. Whole cell lysate was probed for input. Bead lanes contain the protein G-conjugated Sepharose beads used during the immunoprecipitation without the protein input. WB, Western blot. (b) Real-time PCR analysis of the relative expression of cyclin D1 and c-Myc mRNA in NSCLC cells. (c) Western blot analysis of the protein expression of DDX5, (total or nuclear) β-catenin, cyclin D1, and c-Myc. (d) Immunofluorescence analysis of β-catenin in DDX5 or vector-infected NSCLC cells. White arrows indicate nuclear localization of β-catenin. (e, f) Indicated cells transfected with pGL3-cyclinD1 (firefly), TOPflash (TOP) or FOPflash (FOP), and pRL-TK (Renilla) plasmids were subjected to dual luciferase reporter assays 48 h after transfection. Reporter activity was normalized by Renilla luciferase activity. *P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4638002&req=5

fig06: DDX5 promotes β-catenin nuclear accumulation and activates cyclin D1 and c-Myc transcription. (a) Co-immunoprecipitation (IP) analysis of DDX5 and β-catenin in H520 and A549 non-small-cell lung cancer (NSCLC) cells. β-catenin and DDX5 can be reciprocally co-immunoprecipitated by their antibodies. Whole cell lysate was probed for input. Bead lanes contain the protein G-conjugated Sepharose beads used during the immunoprecipitation without the protein input. WB, Western blot. (b) Real-time PCR analysis of the relative expression of cyclin D1 and c-Myc mRNA in NSCLC cells. (c) Western blot analysis of the protein expression of DDX5, (total or nuclear) β-catenin, cyclin D1, and c-Myc. (d) Immunofluorescence analysis of β-catenin in DDX5 or vector-infected NSCLC cells. White arrows indicate nuclear localization of β-catenin. (e, f) Indicated cells transfected with pGL3-cyclinD1 (firefly), TOPflash (TOP) or FOPflash (FOP), and pRL-TK (Renilla) plasmids were subjected to dual luciferase reporter assays 48 h after transfection. Reporter activity was normalized by Renilla luciferase activity. *P < 0.05.

Mentions: As DDX5 was reported to participate in the β-catenin signaling pathway in regulating biological behaviors of cancer cells,10,14 we examined whether DDX5-induced cell proliferation in NSCLC was mediated by this signaling pathway. Co-immunoprecipitation assay indicated a direct physical interaction between DDX5 and β-catenin in both H520 and A549 cells (Fig.6a). We also found that both transcriptional and translational levels of β-catenin target genes cyclin D1 and c-Myc were upregulated by DDX5 overexpression, but suppressed by DDX5 silencing, compared with that in control cells (Figs6b,c,S1). Interestingly, we observed that nuclear accumulation of β-catenin was significantly increased (or decreased) by DDX5 overexpression (or downregulation), whereas the expression of total β-catenin was not affected (Figs6c,d,S1). Moreover, the dual luciferase reporter assay revealed that β-catenin/TCF and cyclin D1 promoter activity were significantly increased in the DDX5-overexpressing NSCLC cells, but decreased in the DDX5 silencing cells (Fig.6e,f). These data collectively suggested that DDX5 might promote NSCLC cell proliferation by activating the β-catenin signaling pathway.


DDX5 promotes proliferation and tumorigenesis of non-small-cell lung cancer cells by activating β-catenin signaling pathway.

Wang Z, Luo Z, Zhou L, Li X, Jiang T, Fu E - Cancer Sci. (2015)

DDX5 promotes β-catenin nuclear accumulation and activates cyclin D1 and c-Myc transcription. (a) Co-immunoprecipitation (IP) analysis of DDX5 and β-catenin in H520 and A549 non-small-cell lung cancer (NSCLC) cells. β-catenin and DDX5 can be reciprocally co-immunoprecipitated by their antibodies. Whole cell lysate was probed for input. Bead lanes contain the protein G-conjugated Sepharose beads used during the immunoprecipitation without the protein input. WB, Western blot. (b) Real-time PCR analysis of the relative expression of cyclin D1 and c-Myc mRNA in NSCLC cells. (c) Western blot analysis of the protein expression of DDX5, (total or nuclear) β-catenin, cyclin D1, and c-Myc. (d) Immunofluorescence analysis of β-catenin in DDX5 or vector-infected NSCLC cells. White arrows indicate nuclear localization of β-catenin. (e, f) Indicated cells transfected with pGL3-cyclinD1 (firefly), TOPflash (TOP) or FOPflash (FOP), and pRL-TK (Renilla) plasmids were subjected to dual luciferase reporter assays 48 h after transfection. Reporter activity was normalized by Renilla luciferase activity. *P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4638002&req=5

fig06: DDX5 promotes β-catenin nuclear accumulation and activates cyclin D1 and c-Myc transcription. (a) Co-immunoprecipitation (IP) analysis of DDX5 and β-catenin in H520 and A549 non-small-cell lung cancer (NSCLC) cells. β-catenin and DDX5 can be reciprocally co-immunoprecipitated by their antibodies. Whole cell lysate was probed for input. Bead lanes contain the protein G-conjugated Sepharose beads used during the immunoprecipitation without the protein input. WB, Western blot. (b) Real-time PCR analysis of the relative expression of cyclin D1 and c-Myc mRNA in NSCLC cells. (c) Western blot analysis of the protein expression of DDX5, (total or nuclear) β-catenin, cyclin D1, and c-Myc. (d) Immunofluorescence analysis of β-catenin in DDX5 or vector-infected NSCLC cells. White arrows indicate nuclear localization of β-catenin. (e, f) Indicated cells transfected with pGL3-cyclinD1 (firefly), TOPflash (TOP) or FOPflash (FOP), and pRL-TK (Renilla) plasmids were subjected to dual luciferase reporter assays 48 h after transfection. Reporter activity was normalized by Renilla luciferase activity. *P < 0.05.
Mentions: As DDX5 was reported to participate in the β-catenin signaling pathway in regulating biological behaviors of cancer cells,10,14 we examined whether DDX5-induced cell proliferation in NSCLC was mediated by this signaling pathway. Co-immunoprecipitation assay indicated a direct physical interaction between DDX5 and β-catenin in both H520 and A549 cells (Fig.6a). We also found that both transcriptional and translational levels of β-catenin target genes cyclin D1 and c-Myc were upregulated by DDX5 overexpression, but suppressed by DDX5 silencing, compared with that in control cells (Figs6b,c,S1). Interestingly, we observed that nuclear accumulation of β-catenin was significantly increased (or decreased) by DDX5 overexpression (or downregulation), whereas the expression of total β-catenin was not affected (Figs6c,d,S1). Moreover, the dual luciferase reporter assay revealed that β-catenin/TCF and cyclin D1 promoter activity were significantly increased in the DDX5-overexpressing NSCLC cells, but decreased in the DDX5 silencing cells (Fig.6e,f). These data collectively suggested that DDX5 might promote NSCLC cell proliferation by activating the β-catenin signaling pathway.

Bottom Line: The DEAD-box-protein DDX5 is an ATP-dependent RNA helicase that is frequently overexpressed in various cancers and acts as a transcriptional co-activator of several transcription factors, including β-catenin.DDX5 is reported to be involved in cancer progression by promoting cell proliferation and epithelial-mesenchymal transition.We found that DDX5 was significantly overexpressed in NSCLC tissues as compared with the matched normal adjacent tissues.

View Article: PubMed Central - PubMed

Affiliation: Department of Thoracic Surgery, Tangdu Hospital, The Fourth Military Medical University, Xi'an, China.

No MeSH data available.


Related in: MedlinePlus