Novel mechanism of reduced proliferation in ovarian clear cell carcinoma cells: Cytoplasmic sequestration of CDK2 by p27

Hiroaki Itamochi, Tomokazu Yoshida, Cheryl Lyn Walker, Chandra Bartholomeusz, Daisuke Aoki, Hideki Ishihara, Nao Suzuki, Junzo Kigawa, Naoki Terakawa, Naoto T. Ueno

Research output: Contribution to journalArticle

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Abstract

Objective: Ovarian clear cell carcinoma (CCC) carries a poor prognosis because of its insensitivity to chemotherapy. We previously found an association between reduced proliferation of CCC and chemoresistance; here we investigated the mechanism of the reduced proliferation. Methods: We assessed cell cycle function by measuring the activity of cyclin-dependent kinases (CDKs) and the protein expression of cyclins, the CDK inhibitors, and p53 in 22 ovarian cancer cell lines and 60 human ovarian cancer specimens. We examined the cellular location of p27, p27 phosphorylated at threonine 157 (p27Thr157), and CDK2 protein by confocal microscopy and western blotting. We tested the effect of the inhibitor of phosphatidylinositol-3-kinase (PI3K) and small interfering RNA against p27 (si-p27) in two CCC cell lines (RMG-I, SMOV-2). Results: CCC cells had lower CDK2 activity and higher p27 expression than serous adenocarcinoma (SA) cells. Low CDK2 activity correlated with high p27 protein expression. p27Thr157 sequestered CDK2 in the cytoplasm, but PI3K inhibitor or si-p27 maintained CDK2 in the nucleus and restored its activity. In human specimens, CDK2 was mostly in the cytoplasm and was spatially associated with p27; CDK2 activity was lower in the CCC than in the SA specimens. si-p27 enhanced the cytotoxic effect of cisplatin, doxorubicin, and gemcitabine in both RMG-I cells and SMOV-2 cells. Conclusions: Reduced CDK2 activity via the cytoplasmic sequestration of CDK2 by p27Thr157 may contribute to suppression of CCC proliferation. A prospective study is needed to determine whether the cytoplasmic sequestration of CDK2 results in the chemoresistance of CCC.

Original languageEnglish
Pages (from-to)641-647
Number of pages7
JournalGynecologic Oncology
Volume122
Issue number3
DOIs
Publication statusPublished - 2011 Sep

Fingerprint

Carcinoma
Threonine
Phosphatidylinositol 3-Kinase
Cyclin-Dependent Kinases
gemcitabine
Ovarian Neoplasms
Cytoplasm
Adenocarcinoma
Cell Proliferation
Cyclin-Dependent Kinase 2
Cell Line
Cyclins
Confocal Microscopy
Doxorubicin
Small Interfering RNA
Cisplatin
Cell Cycle
Western Blotting
Prospective Studies
Drug Therapy

Keywords

  • CDK2
  • Cell cycle
  • Clear cell
  • Ovarian carcinoma
  • p27

ASJC Scopus subject areas

  • Obstetrics and Gynaecology
  • Oncology

Cite this

Novel mechanism of reduced proliferation in ovarian clear cell carcinoma cells : Cytoplasmic sequestration of CDK2 by p27. / Itamochi, Hiroaki; Yoshida, Tomokazu; Walker, Cheryl Lyn; Bartholomeusz, Chandra; Aoki, Daisuke; Ishihara, Hideki; Suzuki, Nao; Kigawa, Junzo; Terakawa, Naoki; Ueno, Naoto T.

In: Gynecologic Oncology, Vol. 122, No. 3, 09.2011, p. 641-647.

Research output: Contribution to journalArticle

Itamochi, H, Yoshida, T, Walker, CL, Bartholomeusz, C, Aoki, D, Ishihara, H, Suzuki, N, Kigawa, J, Terakawa, N & Ueno, NT 2011, 'Novel mechanism of reduced proliferation in ovarian clear cell carcinoma cells: Cytoplasmic sequestration of CDK2 by p27', Gynecologic Oncology, vol. 122, no. 3, pp. 641-647. https://doi.org/10.1016/j.ygyno.2011.05.003
Itamochi, Hiroaki ; Yoshida, Tomokazu ; Walker, Cheryl Lyn ; Bartholomeusz, Chandra ; Aoki, Daisuke ; Ishihara, Hideki ; Suzuki, Nao ; Kigawa, Junzo ; Terakawa, Naoki ; Ueno, Naoto T. / Novel mechanism of reduced proliferation in ovarian clear cell carcinoma cells : Cytoplasmic sequestration of CDK2 by p27. In: Gynecologic Oncology. 2011 ; Vol. 122, No. 3. pp. 641-647.
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abstract = "Objective: Ovarian clear cell carcinoma (CCC) carries a poor prognosis because of its insensitivity to chemotherapy. We previously found an association between reduced proliferation of CCC and chemoresistance; here we investigated the mechanism of the reduced proliferation. Methods: We assessed cell cycle function by measuring the activity of cyclin-dependent kinases (CDKs) and the protein expression of cyclins, the CDK inhibitors, and p53 in 22 ovarian cancer cell lines and 60 human ovarian cancer specimens. We examined the cellular location of p27, p27 phosphorylated at threonine 157 (p27Thr157), and CDK2 protein by confocal microscopy and western blotting. We tested the effect of the inhibitor of phosphatidylinositol-3-kinase (PI3K) and small interfering RNA against p27 (si-p27) in two CCC cell lines (RMG-I, SMOV-2). Results: CCC cells had lower CDK2 activity and higher p27 expression than serous adenocarcinoma (SA) cells. Low CDK2 activity correlated with high p27 protein expression. p27Thr157 sequestered CDK2 in the cytoplasm, but PI3K inhibitor or si-p27 maintained CDK2 in the nucleus and restored its activity. In human specimens, CDK2 was mostly in the cytoplasm and was spatially associated with p27; CDK2 activity was lower in the CCC than in the SA specimens. si-p27 enhanced the cytotoxic effect of cisplatin, doxorubicin, and gemcitabine in both RMG-I cells and SMOV-2 cells. Conclusions: Reduced CDK2 activity via the cytoplasmic sequestration of CDK2 by p27Thr157 may contribute to suppression of CCC proliferation. A prospective study is needed to determine whether the cytoplasmic sequestration of CDK2 results in the chemoresistance of CCC.",
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T1 - Novel mechanism of reduced proliferation in ovarian clear cell carcinoma cells

T2 - Cytoplasmic sequestration of CDK2 by p27

AU - Itamochi, Hiroaki

AU - Yoshida, Tomokazu

AU - Walker, Cheryl Lyn

AU - Bartholomeusz, Chandra

AU - Aoki, Daisuke

AU - Ishihara, Hideki

AU - Suzuki, Nao

AU - Kigawa, Junzo

AU - Terakawa, Naoki

AU - Ueno, Naoto T.

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N2 - Objective: Ovarian clear cell carcinoma (CCC) carries a poor prognosis because of its insensitivity to chemotherapy. We previously found an association between reduced proliferation of CCC and chemoresistance; here we investigated the mechanism of the reduced proliferation. Methods: We assessed cell cycle function by measuring the activity of cyclin-dependent kinases (CDKs) and the protein expression of cyclins, the CDK inhibitors, and p53 in 22 ovarian cancer cell lines and 60 human ovarian cancer specimens. We examined the cellular location of p27, p27 phosphorylated at threonine 157 (p27Thr157), and CDK2 protein by confocal microscopy and western blotting. We tested the effect of the inhibitor of phosphatidylinositol-3-kinase (PI3K) and small interfering RNA against p27 (si-p27) in two CCC cell lines (RMG-I, SMOV-2). Results: CCC cells had lower CDK2 activity and higher p27 expression than serous adenocarcinoma (SA) cells. Low CDK2 activity correlated with high p27 protein expression. p27Thr157 sequestered CDK2 in the cytoplasm, but PI3K inhibitor or si-p27 maintained CDK2 in the nucleus and restored its activity. In human specimens, CDK2 was mostly in the cytoplasm and was spatially associated with p27; CDK2 activity was lower in the CCC than in the SA specimens. si-p27 enhanced the cytotoxic effect of cisplatin, doxorubicin, and gemcitabine in both RMG-I cells and SMOV-2 cells. Conclusions: Reduced CDK2 activity via the cytoplasmic sequestration of CDK2 by p27Thr157 may contribute to suppression of CCC proliferation. A prospective study is needed to determine whether the cytoplasmic sequestration of CDK2 results in the chemoresistance of CCC.

AB - Objective: Ovarian clear cell carcinoma (CCC) carries a poor prognosis because of its insensitivity to chemotherapy. We previously found an association between reduced proliferation of CCC and chemoresistance; here we investigated the mechanism of the reduced proliferation. Methods: We assessed cell cycle function by measuring the activity of cyclin-dependent kinases (CDKs) and the protein expression of cyclins, the CDK inhibitors, and p53 in 22 ovarian cancer cell lines and 60 human ovarian cancer specimens. We examined the cellular location of p27, p27 phosphorylated at threonine 157 (p27Thr157), and CDK2 protein by confocal microscopy and western blotting. We tested the effect of the inhibitor of phosphatidylinositol-3-kinase (PI3K) and small interfering RNA against p27 (si-p27) in two CCC cell lines (RMG-I, SMOV-2). Results: CCC cells had lower CDK2 activity and higher p27 expression than serous adenocarcinoma (SA) cells. Low CDK2 activity correlated with high p27 protein expression. p27Thr157 sequestered CDK2 in the cytoplasm, but PI3K inhibitor or si-p27 maintained CDK2 in the nucleus and restored its activity. In human specimens, CDK2 was mostly in the cytoplasm and was spatially associated with p27; CDK2 activity was lower in the CCC than in the SA specimens. si-p27 enhanced the cytotoxic effect of cisplatin, doxorubicin, and gemcitabine in both RMG-I cells and SMOV-2 cells. Conclusions: Reduced CDK2 activity via the cytoplasmic sequestration of CDK2 by p27Thr157 may contribute to suppression of CCC proliferation. A prospective study is needed to determine whether the cytoplasmic sequestration of CDK2 results in the chemoresistance of CCC.

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KW - Clear cell

KW - Ovarian carcinoma

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