Effect of the cyclin-dependent kinases inhibitor p27 on resistance of ovarian cancer multicellular spheroids to anticancer chemotherapy

Xing, Hui; Wang, Shixuan; Hu, Ke–Qin; Tao, Wenming; Li, Jing; Gao, Qinglai; Yang, Xiaokui; Weng, Danhui; Lu, Ye; Ma, Ding

doi:10.1007/s00432-005-0677-9

Cited by 37 publications

(36 citation statements)

References 25 publications

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“…It is supposed that p27 in nucleus binds to cyclin E-CDK2 complex and inactivates the complex in G0/G1 phase, but once p27 is transported to cytosol and degraded, the complex is activated and it triggers the transition from G1 to S phase. Our data were consistent with this kinetics of p27, so it was suggested that intranuclear p27 plays a central role for cellcycle quiescence in sphere forming cells (20)(21)(22).…”

Section: Discussionsupporting

confidence: 90%

Analogy between sphere forming ability and stemness of human hepatoma cells

Tanaka¹

2010

Oncol Rep

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Abstract. Increasing evidence suggests that cancers contain a small subset of cancer-initiating cells, so-called cancer stem cells (CSCs) that are capable of regenerating a tumor after chemoradiation therapy. Sphere forming ability is known to be one of properties of CSCs, but the significance remains unclear. The present study focused on sphere formation of human hepatoma cells in three-dimensional culture in order to evaluate the analogy between sphere forming ability and stemness of cancer cells in vitro. Under three-dimensional culture condition, HepG2, Hep3B and PLC/PRF/5 cells demonstrated the sphere formation while SK-Hep1 and Huh-7 cells did not. The population of G0/G1 phase increased in the spheres compared with the monolayer (67 vs. 38%). In spite of no significant difference in stem cell surface markers (CD44, CD90, CD133, EpCAM and ABCG2), remarkable upregulation of p27 CDK inhibitor was observed in sphere forming cells. Immunofluorescence analysis revealed the nuclear expression of p27 in the whole of the sphere, but weak expression of p21 only at the peripheral area. The spheres acquired chemoresistance to cisplatin compared with the monolayers (58.9 vs. 16.2 μM in IC50). This model was useful for assessment of the role of cell-cycle quiescence in the stemness and chemoresistance of cancer cells. IntroductionThere is an emerging concept of cancer stem cells (CSCs) that cancer cells do not consist of homogeneous population but include a small subpopulation of cells having the ability of self-renewal and differentiation into multiple phenotypes (1). Several unique properties of CSCs have been identified including drug-efflux ability (side population) (2,3), maintenance of quiescence, sphere formation, high tumorigenicity, and resistance to hypoxia and chemoradiation (4). After anticancer treatment that kills most cancer cells, such drugresistant CSCs might survive and finally generate new populations, resulting in cancer recurrence and metastasis. Detailed analysis on biological characteristics of CSCs are required to overcome the resistance of cancer.Recent studies revealed that the sphere formation might be essential for cancer-initiating ability of CSCs (5-7), but its significance and mechanism remain unclear. The sphereforming cells in human hepatoma were reported to associate with the expression of stemness markers such as CD90 (8,9), CD133 (10,11), EpCAM (12), ABCG2 (13) and CD44 (14). In this study, we focused on the sphere formation ability using 3D culture system to evaluate the in vitro analogy to the stemness phenotypes in human hepatoma cells. Materials and methodsCell lines. Human hepatoma cell lines, HepG2, Huh-7, PLC/ PRF/5 and SK-Hep1 were analyzed (15). Additionally, we used p53-deficient hepatoma cell line Hep3B as well as Hep3B transfected with wild-type p53 gene (16); named as Hep3B-p53(-) and Hep3B-p53(+), respectively. Culture media were the recommended media supplemented with 10% fetal bovine serum, 100 U/ml penicillin and 100 μg/ml streptomycin. All cell lines were cultivated...

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Section: Discussionsupporting

confidence: 90%

Analogy between sphere forming ability and stemness of human hepatoma cells

Tanaka¹

2010

Oncol Rep

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“…However, the 2D monolayer in vitro model does not exhibit the full range of drug resistance observed in in vivo solid tumours. For this reason, three-dimensional (3D) multicellular tumour spheroids (MTS) have been used as an in vitro model of solid tumours for drug resistance studies because they mimic the growth characteristics of in vivo tumours more closely than cancer cell line monolayers (5)(6)(7)(8)(9).…”

Section: Introductionmentioning

confidence: 99%

“…The acquired MCR has been demonstrated to be linked to a limitation of drug diffusion, to the activation of survival pathways such as NF-κB or mTOR, or to the upregulation of p27 (5)(6)(7)(8)(9). However, the molecular basis of MCR is not yet clear.…”

Section: Introductionmentioning

confidence: 99%

Implication of necrosis-linked p53 aggregation in acquired apoptotic resistance to 5-FU in MCF-7 multicellular tumour spheroids

Kang¹

2010

Oncol Rep

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Abstract. Three-dimensional (3D) multicellular tumour spheroids (MTS) have been used as an in vitro model of solid tumours for drug resistance studies because they mimic the growth characteristics of in vivo tumours more closely than in vitro two-dimensional (2D) culture of cancer cell lines. As observed in solid tumours, MTS exhibits a proliferation gradient with outer regions consisting of proliferating cells that surround inner quiescent cells. The innermost cells in core regions undergo cell death mostly by necrosis to form necrotic core due to insufficient supply of oxygen and nutrient such as glucose with increasing size of spheroids. Tumour necrosis is thought to indicate a poor prognosis and to contribute to acquisition of chemoresistance in solid tumours; however, the mechanism underlying necrosis-mediated chemoresistance remains unclear. In this study, we examined the chemoresistance to 5-Fluorouracil (5-FU) using MCF-7 breast cancer MTS. 5-FU (400 μM) induced apoptosis in MCF-7 cell monolayer as determined by HO/PI staining, PARP cleavage, p53 induction, Bax induction, and Bcl-2 downregulation. When MCF-7 breast tumour spheroids were cultured on agarose for 8 days, they reached ~700 μm in diameter, with a necrotic core. We found that 5-FU-induced apoptosis is markedly reduced in spheroids that were cultured for 9 days and had necrotic core, compared with MCF-7 monolayer cells and spheroids that were cultured for 6 days and had no necrotic core, indicating that the formation of necrotic core may be linked to acquisition of chemoresistance to 5-FU. We also found that a specific set of cellular proteins including p53 was aggregated into a RIPAinsoluble form during MTS culture. Furthermore, most of p53 induced by 5-FU was aggregated in MTS with necrotic core. Our results suggest that necrosis-linked p53 aggregation may contribute to acquired apoptotic resistance to 5-FU in MTS model system.

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“…Interestingly, p16 INK4b , p21 Cip1 and p27 Kip1 , whose main function is to induce cell cycle arrest and apoptosis, induce autophagy [16]. p27 Kip1 also can down-regulate P-gp [17]. Thus, enhancement of CKI signaling is particularly noteworthy among the putative molecular targets other than COX-2 in CLX-mediated in vitro antiproliferative effects in cancer cells [13,14].…”

Section: Introductionmentioning

confidence: 99%

Down-regulation of the HGF/MET autocrine loop induced by celecoxib and mediated by P-gp in MDR-positive human hepatocellular carcinoma cell line

Mazzanti

Platini

Bottini

et al. 2009

Biochemical Pharmacology

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This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.Page 1 of 33A c c e p t e d M a n u s c r i p t Graphical AbstractThe in vitro pro-autophagic, antiproliferative and pro-apoptotic effect of celecoxib on MDRpositive cells is mediated by the action of P-gp on the HGF/MET autocrine loop. small interfering RNA. These findings demonstrate that the constitutive expression of P-gp is involved in the HGF/MET autocrine loop that leads to increased expression of Bcl-2 and mTor, inhibition of eIF2α expression, resistance to autophagy/apoptosis and progression in the cell cycle.Since mTor inhibitors have been proposed in treatment of "drug resistant" cancer, these data may help explain the reversing effect of mTor inhibitors.

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Effect of the cyclin-dependent kinases inhibitor p27 on resistance of ovarian cancer multicellular spheroids to anticancer chemotherapy

Abstract: Our results implicate that p27 serves as a regulator of drug resistance in ovarian tumors. ASON-mediated alteration of p27 reverses resistance of ovarian cancer to anticancer agents that are associated with increased sensitivity of ovarian cancer cells to chemotherapeutic agents.

Cited by 37 publications

References 25 publications

Analogy between sphere forming ability and stemness of human hepatoma cells

Analogy between sphere forming ability and stemness of human hepatoma cells

Implication of necrosis-linked p53 aggregation in acquired apoptotic resistance to 5-FU in MCF-7 multicellular tumour spheroids

Down-regulation of the HGF/MET autocrine loop induced by celecoxib and mediated by P-gp in MDR-positive human hepatocellular carcinoma cell line

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