European Journal of Obstetrics & Gynecology and Reproductive Biology
Perifosine inhibits growth of human experimental endometrial cancers by blockade of AKT phosphorylation
Introduction
Endometrial carcinoma is the fourth most common cancer and the eighth leading cause of death from malignancy in women. With 40,000 new cases resulting in 6800 deaths every year in the US alone, it is the most common neoplasm of the female genital tract and accounts for almost half of all gynecologic cancers. Lifetime risk of developing endometrial cancer is about 2–3% [1]. Primary surgery followed by radiation has become the most widely accepted treatment for endometrial cancer. In the case of late-stage or recurrent disease systemic, hormonal or chemotherapy are added. Early-stage endometrial cancer carries a good prognosis, with 5-year survival rates of 87% and 76% for FIGO stages I and II, respectively [2]. However, 5-year survival rate drops to 59% (FIGO III) and 18% (FIGO IV) in patients with late-stage disease [2]. Overall survival is poorest in patients with recurrent endometrial cancer (7.7%) [3]. Accordingly, new therapeutic modalities are needed for the treatment of late-stage and recurrent endometrial cancer.
The serine/threonine kinase AKT/PKB pathway presents a promising new target for molecular therapeutics, as it integrates both extracellular and intracellular oncogenic signals. Alterations of the AKT pathway have been detected in a variety of human malignancies including endometrial cancer [4]. AKT has a wide range of downstream targets that regulate tumor-associated cell processes such as cell growth, cell cycle progression, survival, migration, epithelial–mesenchymal transition and angiogenesis [5]. Blockage of AKT signaling results in programmed cell death (PCD) and growth inhibition of tumor cells with elevated AKT. For this reason, AKT has become a promising drug target for cancer therapy.
The PTEN tumor suppressor gene is a central negative regulator of the AKT signaling cascade and the most commonly altered component of the AKT pathway in human malignancies [6]. Mutations in PTEN result in significantly increased AKT activity [7]. Accordingly PTEN mutations occur in 50% of endometrial carcinomas [4] and patients with loss of PTEN expression followed by AKT phosphorylation are burdened with a poor prognosis [8]. Therefore targeting the AKT-pathway may be a favourable strategy in endometrial cancer.
Perifosine is a synthetic alkylphospholipid which is orally active and inhibits AKT activation [9]. AKT is activated by phosphorylation, therefore AKT phosphorylation is a surrogate marker for the activation of this enzyme. In addition to inhibition of AKT in some cell types perifosine downregulates the anti-apoptotic mitogene activated protein kinase extracellular signal regulated kinase (MEK-ERK 1/2) pathway and activates the proapoptotic c-jun-N-kinase (JNK) network, thus modulating the balance between the survival and death signaling cascades, thereby inducing PCD [10], [11]. Perifosine has shown anti-tumor activity in a variety of cancers in vitro[12] and in vivo[13]. So far perifosine was tested in clinical phase II studies in breast [14], head and neck [15], prostate [16], pancreatic [17] cancers and soft tissue sarcoma [18] patients.
The current study investigates a potential anti-tumor activity of perifosine in in vitro models of human endometrial cancers using perifosine either on its own or in combination with an established chemotherapeutic drug.
Section snippets
Reagents and cells
The alkylphospholipid perifosine was kindly provided by AeternaZentarisGmbH (Frankfurt, Germany) and dissolved in DMSO (concentration of the stock solution = 10 mM). HEC 1A and Ishikawa human endometrial cancer cell lines were obtained from American Type Culture Collection (Manassas, VA, USA). HEC 1A cells were grown in RPMI-1640 medium, Ishikawa cells in Dulbecco's modified eagle medium (DMEM) (both from PAA, Cölbe, Germany). Both media were supplemented with 10% fetal calf serum (FCS) (Biochrom,
Perifosine inhibits AKT phosphporylation in Ishikawa and HEC 1A human endometrial cancer cells
The presence of phosphorylated and total AKT was assessed by Western blotting in Ishikawa and HEC 1A cells that had been treated with 0 μM, 10 μM and 40 μM perifosine. Specific antibody reactivity was detected at Mr = 60 kDa for both phosphorylated and total AKT. Phosphorylated AKT was dose-dependently decreased by treatment with perifosine (Fig. 1).
Effects of perifosine on the growth of HEC 1A and Ishikawa human endometrial cancer cells
To investigate whether impaired AKT phosphorylation would lead to reduced growth or to cell death in human endometrial cancer cells, Ishikawa and HEC 1A
Discussion
Treatment of advanced or recurrent endometrial cancer remains a major problem. Radiation therapy is frequently applied, even though the resulting survival benefit is modest at most. Combination chemotherapy shows objective responses in 40–60% of patients, but the survival is less than 1 year. The most commonly used therapeutic regimen combines cisplatin and doxorubicin. Improved response rates, longer progression-free survival and increased overall survival may be achieved by adding paclitaxel
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2015, Advances in Protein Chemistry and Structural BiologyCitation Excerpt :Clinical trials for this combination therapy in MM and neuroblastoma patients are underway (Rossi et al., 2012). Perifosine could be a potent sensitizer of conventional chemotherapy, for example, in MM (Hideshima et al., 2006), glioma (Momota, Nerio, & Holland, 2005), medulloblastoma (Kumar et al., 2009), endometrial cancer (Engel et al., 2008), osteosarcoma (Yao et al., 2013), and leukemias (Nyakern, Cappellini, Mantovani, & Martelli, 2006; Papa et al., 2008). Phase II clinical trials with perifosine in combination with bortezomib and dexamethasone in MM patients (Richardson et al., 2012), or with capecitabine in patients with metastatic colorectal cancer (Bendell et al., 2011) were promising.
Novel alkylphospholipid-DTC hybrids as promising agents against endocrine related cancers acting via modulation of Akt-pathway
2014, European Journal of Medicinal ChemistryCitation Excerpt :Similarly, a dose dependent increase green fluorescence was also observed in DU-145 cells treated with compound 14 (Fig. 5B). It is well-established that alkylphospholipid derivatives like hexadecylphosphocholines miltefosine [20], perifosine [21] and erufosine [22] downregulate Akt phosphorylation thereby blocking Akt survival pathway. Since compounds (14 and 18) are alkylphospholipid derivatives and share considerable homology with the other pharmacologically active compounds of this class, the effect of compound 14 and 18 was tested on phosphorylation of Akt as described in experimental section [23].
Synthesis, characterization and Akt phosphorylation inhibitory activity of cyclopentanecarboxylate-substituted alkylphosphocholines
2013, Bioorganic and Medicinal ChemistryCitation Excerpt :However, its potency in phase II clinical studies was much less.34 Several APC compounds have been synthesized as inhibitors of PI3K/Akt signaling and were tested in a variety of tumor cells.6,9,35–38 However, these existing APCs have moderate potencies compared with other mechanism of antitumor agents, and had some adverse effects.39
Anticancer mechanisms and clinical application of alkylphospholipids
2013, Biochimica et Biophysica Acta - Molecular and Cell Biology of LipidsCitation Excerpt :Clinical trials for this combination therapy in MM and neuroblastoma patients are underway [188,189]. Perifosine could be a potent sensitizer of conventional chemotherapy, for example in MM [103], glioma [102], medulloblastoma [180], endometrial cancer [190], osteosarcoma [191] and leukemias [192,193]. Phase II clinical trials with perifosine in combination with bortezomib and dexamethasone in MM patients [41] or with capecitabine in patients with metastatic colorectal cancer [43] were promising.
The role of the phosphatidylinositol 3-kinase (PI3K) pathway in the development and treatment of uterine cancer
2011, Gynecologic OncologyCitation Excerpt :As has been observed with rapalogs, inhibition of AKT could abrogate negative feedback loops, leading to a paradoxical increase in activity among non-AKT-dependent effectors of the PI3K pathway. In preclinical models, perifosine (KRX-0401) inhibited the growth of experimental models of human uterine cancers by blockade of AKT phosphorylation, and these effects were increased with cisplatin combinations [60]. A Phase I trial of perifosine in soft tissue sarcoma, including uterine, did not show evidence of activity [61].
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These authors contributed equally.