An activated mTOR/p70S6K signaling pathway in esophageal squamous cell carcinoma cell lines and inhibition of the pathway by rapamycin and siRNA against mTOR

Hou, Guoqing; Xue, Liyan; Lu, Zhaoming; Fan, Tingting; Tian, Fang; Xue, Yanli

doi:10.1016/j.canlet.2007.01.026

Cited by 125 publications

(94 citation statements)

References 36 publications

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“…After 24 h, one plate was subjected to cell proliferation assay. In the proliferation assay, cells were placed in culture medium (RPMI 1640 with 10% fetal calf serum) with 10:1 Cell Counting Kit-8 (beyotime company, China) 17 and incubated at 37°C, 5% CO 2 for 3 h. Finally, A450 nm of each well was measured using an ELISA plate reader (Thermo Labsystems).…”

Section: Methodsmentioning

confidence: 99%

Pin1 expression contributes to lung cancer prognosis and carcinogenesis

Tan¹,

Zhou²,

Wan³

et al. 2010

Cancer Biology & Therapy

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Section: Methodsmentioning

confidence: 99%

Pin1 expression contributes to lung cancer prognosis and carcinogenesis

Tan¹,

Zhou²,

Wan³

et al. 2010

Cancer Biology & Therapy

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“…Then 10 lL WST-8 dye was added to each well and incubated at 37 C for 4 hr. Finally, the absorbance of each well was determined at 450 nm using a microplate reader (Bio-Tek instruments) (Hou et al, 2007). The percentages of surviving cells from each group relative to control were defined as proliferation rate, using the following formula: % proliferation rate ¼ A450 of treated cells/A450 of control cells Â 100%.…”

Section: Cell Proliferation Assaymentioning

confidence: 99%

Metformin Induces G1 Cell Cycle Arrest and Inhibits Cell Proliferation in Nasopharyngeal Carcinoma Cells

Zhao

Wen

Jia

et al. 2011

The Anatomical Record

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It has been reported that metformin, a biguanide derivative widely used in type II diabetic patients, has antitumor activities in some cancers by activation of AMP-activated protein kinase (AMPK). But its role in nasopharyngeal carcinoma (NPC) is not known. Here, we reported for the first time that 1-50 mM of metformin in a dose-and time-dependent manner suppressed cell proliferation and colony formation in NPC cell line, C666-1. Further studies revealed that the protein level of cyclin D1 decreased and the percentage of the cells in G0/G1 phase increased by 5 mM metformin treatment. Metformin also induced the phosphorylation of AMPK (T172) in a time-dependent manner. Mammalian target of rapamycin complex 1 (mTORC1), which is negatively regulated by AMPK and plays a central role in cell growth and proliferation, was inhibited by metformin, as manifested by dephosphorylation of its downstream targets 40S ribosomal S6 kinase 1 (S6K1) (T389), the eukaryotic translation initiation factor 4E (eIF4E)-binding protein 1 (4E-BP1) (T37/46) and S6 (S235/236) in C666-1 cells. In a summary, metformin prevents proliferation of C666-1 cells by down-regulating cyclin D1 level and inducing G1 cell cycle arrest. AMPK-mediated inhibition of mTORC1 signaling may be involved in this process. Anat Rec, 294:1337Rec, 294: -1343Rec, 294: , 2011. V V C 2011 Wiley-Liss, Inc.

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“…Western blot was performed according to published methods (22). Briefly, EC9706 cells untreated and transfected with Pim-1 siRNA and control siRNA were harvested at 48 h after treatment and lysed for 20 min in cold lysis buffer.…”

Section: Semi-quantitative Rt-pcrmentioning

confidence: 99%

“…The findings presented herein will provide the first new data for future therapy of ESCC. supplemented with 10% fetal bovine serum (FBS) (HyClone Laboratories, Logan, USA), 100 U/ml penicillin and 100 μg/ml streptomycin at 37˚C in the presence of 5% CO 2 , as described in the previous studies from our laboratory (21,22 Cell proliferation assay. Cell proliferation was determined using WST-8 dye (Beyotime Institute of Biotechnology, China) according to manufacturer's instructions.…”

Section: Introductionmentioning

confidence: 99%

A pivotal role for Pim-1 kinase in esophageal squamous cell carcinoma involving cell apoptosis induced by reducing Akt phosphorylation

Yu²,

Zhang³

et al. 2010

Oncol Rep

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Abstract. It is well documented that Provirus integration site for Moloney murine leukemia virus 1 (Pim-1), as a protooncogene encoding a serine/threonine kinase with multiple cellular functions, is tightly associated with the occurrence and development of tumors. Overexpression of Pim-1 plays a critical role in the progression of several different tumors. In this study, esophageal squamous cell carcinoma (ESCC) EC9706 cells with Pim-1 siRNA treatment resulted in a clear decrease of Pim-1 levels, followed by inhibiting cell proliferation and inducing apoptosis. Further, Pim-1 siRNA reduced phosphorylation of Akt and Bad, and increased cleaved caspase-3/-9 activities and expression levels. These data suggest that Pim-1 siRNA-mediated apoptosis is closely related to the decrease in Akt and Bad phosphorylation and increase in cleaved caspase-3/-9 activities, and thus manipulation of Pim-1 is a potential target for molecular therapy in the clinical treatment of patients with ESCC. IntroductionEsophageal squamous cell carcinoma (ESCC), the major histological form of esophageal cancer, is the sixth most frequent cause of cancer deaths worldwide (1). ESCC shows a poor prognosis due to the lack of early screening strategy and is often presented in an advanced stage at the time of diagnosis (2). ESCC is one of the most frequently diagnosed cancers in developing countries, especially in China (3). Currently, although therapy strategies have been improved, the prognosis of patients with ESCC is still poor and 5-year survival rate for ESCC appears a dramatic decrease (4). Obviously, there is a clear need to seek for new molecular prognostic markers that helps identify patients at a higher risk of death and improve the prognosis of patients with ESCC.Provirus integration site for Moloney murine leukemia virus 1 (Pim-1) was originally identified as a preferential proviral integration site in Moloney murine leukemia virusinduced T cell lymphomas (5) and the first described member of the Pim kinase family, including another two serine/ threonine kinases Pim-2 and Pim-3 (6). More and more evidence has demonstrated that Pim-1, a proto-oncogene encoding a serine/threonine protein kinase, plays an essential role in cell survival, cell proliferation, cell differentiation, cell cycle, and cell apoptosis (7-14), especially in the occurrence and development of tumors (15)(16)(17)(18)(19)(20). Therefore, the rapidly increasing knowledge on the role of the Pim-1 in cancer will hopefully provide useful molecular markers for detection and prognosis of tumor.However, the role for Pim-1 in ESCC has not yet been reported. Therefore, the purpose of this study was to analyze the effect of Pim-1 siRNA on Pim-1 expression levels in ESCC cell line EC9706 cells by RT-PCR and Western blotting methods, and subsequently, to investigate the effect of Pim-1 siRNA on cell proliferation and apoptosis in EC9706 cells in vitro. Further, Akt and Bad phosphorylation levels, caspase-3/-9 activities, and cleaved caspase-3/-9 expression levels were investigated ...

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An activated mTOR/p70S6K signaling pathway in esophageal squamous cell carcinoma cell lines and inhibition of the pathway by rapamycin and siRNA against mTOR

Cited by 125 publications

References 36 publications

Pin1 expression contributes to lung cancer prognosis and carcinogenesis

Pin1 expression contributes to lung cancer prognosis and carcinogenesis

Metformin Induces G1 Cell Cycle Arrest and Inhibits Cell Proliferation in Nasopharyngeal Carcinoma Cells

A pivotal role for Pim-1 kinase in esophageal squamous cell carcinoma involving cell apoptosis induced by reducing Akt phosphorylation

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