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Stathmin1 regulates p27 expression, proliferation and drug resistance, resulting in poor clinical prognosis in cholangiocarcinoma.

Watanabe A, Suzuki H, Yokobori T, Tsukagoshi M, Altan B, Kubo N, Suzuki S, Araki K, Wada S, Kashiwabara K, Hosouchi Y, Kuwano H - Cancer Sci. (2014)

Bottom Line: Patients in the high-STMN1-expression group were associated with shorter recurrence-free survival and overall survival than those in the low-expression group.STMN1 knockdown inhibited proliferation and increased the sensitivity of EHCC cells to paclitaxel.Understanding the regulation of p27 by STMN1 could provide new insights for overcoming therapeutic resistance in EHCC.

View Article: PubMed Central - PubMed

Affiliation: Department of General Surgical Science, Gunma University Graduate School of Medicine, Gunma, Japan.

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(a) Stathmin1 (STMN1) bound to p27 in the cytoplasm of HuCCT1 cells. Primary STMN1 rabbit and p27 mouse antibodies bound to STMN1 and p27 complexes were combined with secondary proximity ligation assay probes. The interaction events are visible as red dots (nuclear staining with DAPI). (b) siRNA-mediated STMN1 suppression. STMN1 protein levels were measured using western blotting after transfection with STMN1 siRNA. p27 expression in STMN1 siRNA-transfected and untreated HuCCT-1 cells was assessed using western blotting of total and nuclear protein. β-Actin was used as a loading control for total protein and Histone H1 was used as a loading control for nuclear protein. (c) HuCCT-1 cells were transfected with STMN1 siRNA and WST-8 proliferative assays were performed and compared with untransfected cells. (d) Paclitaxel sensitivity was determined using a WST-8 assay. STMN1 siRNA transfection significantly increased paclitaxel sensitivity compared with untransfected cells. Each time point represents the mean ± SD of triplicate determinations in two independent experiments. *P < 0.05.
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fig03: (a) Stathmin1 (STMN1) bound to p27 in the cytoplasm of HuCCT1 cells. Primary STMN1 rabbit and p27 mouse antibodies bound to STMN1 and p27 complexes were combined with secondary proximity ligation assay probes. The interaction events are visible as red dots (nuclear staining with DAPI). (b) siRNA-mediated STMN1 suppression. STMN1 protein levels were measured using western blotting after transfection with STMN1 siRNA. p27 expression in STMN1 siRNA-transfected and untreated HuCCT-1 cells was assessed using western blotting of total and nuclear protein. β-Actin was used as a loading control for total protein and Histone H1 was used as a loading control for nuclear protein. (c) HuCCT-1 cells were transfected with STMN1 siRNA and WST-8 proliferative assays were performed and compared with untransfected cells. (d) Paclitaxel sensitivity was determined using a WST-8 assay. STMN1 siRNA transfection significantly increased paclitaxel sensitivity compared with untransfected cells. Each time point represents the mean ± SD of triplicate determinations in two independent experiments. *P < 0.05.

Mentions: To examine the protein complexes of STMN1 and p27 in HuCCT1 cells, we performed PLA, which revealed the complexes as red spots in the cytoplasm (Fig.3a). Thus, STMN1 interacts with p27 in the cytoplasm, but not in the nucleus, of EHCC cells.


Stathmin1 regulates p27 expression, proliferation and drug resistance, resulting in poor clinical prognosis in cholangiocarcinoma.

Watanabe A, Suzuki H, Yokobori T, Tsukagoshi M, Altan B, Kubo N, Suzuki S, Araki K, Wada S, Kashiwabara K, Hosouchi Y, Kuwano H - Cancer Sci. (2014)

(a) Stathmin1 (STMN1) bound to p27 in the cytoplasm of HuCCT1 cells. Primary STMN1 rabbit and p27 mouse antibodies bound to STMN1 and p27 complexes were combined with secondary proximity ligation assay probes. The interaction events are visible as red dots (nuclear staining with DAPI). (b) siRNA-mediated STMN1 suppression. STMN1 protein levels were measured using western blotting after transfection with STMN1 siRNA. p27 expression in STMN1 siRNA-transfected and untreated HuCCT-1 cells was assessed using western blotting of total and nuclear protein. β-Actin was used as a loading control for total protein and Histone H1 was used as a loading control for nuclear protein. (c) HuCCT-1 cells were transfected with STMN1 siRNA and WST-8 proliferative assays were performed and compared with untransfected cells. (d) Paclitaxel sensitivity was determined using a WST-8 assay. STMN1 siRNA transfection significantly increased paclitaxel sensitivity compared with untransfected cells. Each time point represents the mean ± SD of triplicate determinations in two independent experiments. *P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4317896&req=5

fig03: (a) Stathmin1 (STMN1) bound to p27 in the cytoplasm of HuCCT1 cells. Primary STMN1 rabbit and p27 mouse antibodies bound to STMN1 and p27 complexes were combined with secondary proximity ligation assay probes. The interaction events are visible as red dots (nuclear staining with DAPI). (b) siRNA-mediated STMN1 suppression. STMN1 protein levels were measured using western blotting after transfection with STMN1 siRNA. p27 expression in STMN1 siRNA-transfected and untreated HuCCT-1 cells was assessed using western blotting of total and nuclear protein. β-Actin was used as a loading control for total protein and Histone H1 was used as a loading control for nuclear protein. (c) HuCCT-1 cells were transfected with STMN1 siRNA and WST-8 proliferative assays were performed and compared with untransfected cells. (d) Paclitaxel sensitivity was determined using a WST-8 assay. STMN1 siRNA transfection significantly increased paclitaxel sensitivity compared with untransfected cells. Each time point represents the mean ± SD of triplicate determinations in two independent experiments. *P < 0.05.
Mentions: To examine the protein complexes of STMN1 and p27 in HuCCT1 cells, we performed PLA, which revealed the complexes as red spots in the cytoplasm (Fig.3a). Thus, STMN1 interacts with p27 in the cytoplasm, but not in the nucleus, of EHCC cells.

Bottom Line: Patients in the high-STMN1-expression group were associated with shorter recurrence-free survival and overall survival than those in the low-expression group.STMN1 knockdown inhibited proliferation and increased the sensitivity of EHCC cells to paclitaxel.Understanding the regulation of p27 by STMN1 could provide new insights for overcoming therapeutic resistance in EHCC.

View Article: PubMed Central - PubMed

Affiliation: Department of General Surgical Science, Gunma University Graduate School of Medicine, Gunma, Japan.

Show MeSH
Related in: MedlinePlus