Knockdown of stat3 expression by RNAi inhibits in vitro growth of human ovarian cancer

Zhao, Shuhua; Zhao, Fan; Zheng, Jingying; Gao, Lifang; Zhao, Xin; Cui, Manhua

doi:10.2478/v10019-011-0013-8

Cited by 22 publications

(12 citation statements)

References 27 publications

(32 reference statements)

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“…It was reported that the suppression of Cyclin D1 with antisense induces both cell growth arrest and apoptosis [44,45]. Also, it was shown that the knockdown of STAT3 expression by RNA interference (RNAi) suppresses the growth of SKOV3 cells and arrests the cell cycle in the G1 phase [46]. Consistent with our findings, the authors mentioned that “cells transfected with siRNA-stat3 became less confluent and some cells were rounded and detached from the plates, when compared with cells transfected with scrambled vectors”.…”

Section: Discussionmentioning

confidence: 99%

Tagging of Genomic STAT3 and STAT1 with Fluorescent Proteins and Insertion of a Luciferase Reporter in the Cyclin D1 Gene Provides a Modified A549 Cell Line to Screen for Selective STAT3 Inhibitors

Samsonov¹,

Zenser²,

Zhang³

et al. 2013

PLoS ONE

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Signal transducer and activator of transcription 3 (STAT3) is an oncogenic protein that is constitutively activated in numerous cancer cell lines and human cancers. Another STAT family member, STAT1, possesses cancer-inhibitory properties and can promote apoptosis in tumor cells upon activation. To better characterize these important cancer related genes, we tagged STAT3 and STAT1 loci with fluorescent protein (FP) sequences (RFP and GFP respectively) by targeted integration via zinc finger nuclease (ZFN) - mediated homologous recombination in A549 cells that express aberrantly activated STAT3. We inserted the FP transgenes at the N-terminus of the STAT3 locus and at the C-terminus of the STAT1 locus. The integration resulted in endogenous expression of fluorescent STAT3 and STAT1 chimeric fusion proteins. When stimulated with IL-6 or IFN-γ, the cells showed robust nuclear translocation of RFP-STAT3 or STAT1-GFP, respectively. Pre-incubation of cells with a known specific STAT3 inhibitor showed that IFN-γ-induced translocation of STAT1-GFP was not impaired. STAT3 activates multiple downstream targets such as genes involved in cell cycle progression - e.g. cyclin D1. To detect changes in expression of endogenous cyclin D1, we used ZFN technology to insert a secreted luciferase reporter behind the cyclin D1 promoter and separated the luciferase and cyclin D1 coding regions by a 2A sequence to induce a translational skip. The luciferase insertion was made in the RFP-STAT3/STAT1-GFP cell line to have all three reporters in a single cell line. Addition of a STAT3 inhibitor led to suppression of cyclin D1 promoter activity and cell growth arrest. The triple-modified cell line provides a simple and convenient method for high-content screening and pre-clinical testing of potential STAT3 inhibitors in live cells while ensuring that the STAT1 pathway is not affected. This approach of reporting endogenous gene activities using ZFN technology could be applied to other cancer targets.

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Section: Discussionmentioning

confidence: 99%

Tagging of Genomic STAT3 and STAT1 with Fluorescent Proteins and Insertion of a Luciferase Reporter in the Cyclin D1 Gene Provides a Modified A549 Cell Line to Screen for Selective STAT3 Inhibitors

Samsonov¹,

Zenser²,

Zhang³

et al. 2013

PLoS ONE

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show abstract

“…Inhibition of STAT3 with the orally active JAK inhibitor, AZD1480, was found to decrease tumor growth in neuroblastoma and pediatric sarcomas through downregulation of cell cycle regulators, cyclin D1, cyclin D3 and cdc25A, anti-apoptotic genes Bcl-2 and survivin, the metastasis-related factor TIMP-1, and c-Myc. Knockdown of STAT3 expression by RNAi inhibited the growth of human ovarian cancer through downregulation of cyclin D1, c-Myc and Bcl-2 [145].…”

Section: Proliferationmentioning

confidence: 99%

Targeting the STAT3 signaling pathway in cancer: Role of synthetic and natural inhibitors

Siveen

Sikka

Surana

et al. 2014

Biochimica et Biophysica Acta (BBA) - Reviews on Cancer

478

510

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Signal transducers and activators of transcription (STATs) comprise a family of cytoplasmic transcription factors that mediate intracellular signaling that is usually generated at cell surface receptors and thereby transmit it to the nucleus. Numerous studies have demonstrated constitutive activation of STAT3 in a wide variety of human tumors, including hematological malignancies (leukemias, lymphomas, and multiple myeloma) as well as diverse solid tumors (such as head and neck, breast, lung, gastric, hepatocellular, colorectal and prostate cancers). There is strong evidence to suggest that aberrant STAT3 signaling promotes initiation and progression of human cancers by either inhibiting apoptosis or inducing cell proliferation, angiogenesis, invasion, and metastasis. Suppression of STAT3 activation results in the induction of apoptosis in tumor cells, and accordingly its pharmacological modulation by tyrosine kinase inhibitors, antisense oligonucleotides, decoy nucleotides, dominant negative proteins, RNA interference and chemopreventive agents have been employed to suppress the proliferation of various human cancer cells in culture and tumorigenicity in vivo. However, the identification and development of novel drugs that can target deregulated STAT3 activation effectively remains an important scientific and clinical challenge. This review presents the evidence for critical roles of STAT3 in oncogenesis and discusses the potential for development of novel cancer therapies based on mechanistic understanding of STAT3 signaling cascade.

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“…Persistent activation of STAT3 has been reported in ovarian cancer cells [23]. Research using RNAi technology to knockdown the STAT3 gene in ovarian cancer cells has also been reported [24,25].…”

Section: Introductionmentioning

confidence: 99%

Knockdown of STAT3 expression in SKOV3 cells by biodegradable siRNA–PLGA/CSO conjugate micelles

Zhao

Zheng

Zhang

et al. 2015

Colloids and Surfaces B: Biointerfaces

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Knockdown of stat3 expression by RNAi inhibits in vitro growth of human ovarian cancer

Cited by 22 publications

References 27 publications

Tagging of Genomic STAT3 and STAT1 with Fluorescent Proteins and Insertion of a Luciferase Reporter in the Cyclin D1 Gene Provides a Modified A549 Cell Line to Screen for Selective STAT3 Inhibitors

Tagging of Genomic STAT3 and STAT1 with Fluorescent Proteins and Insertion of a Luciferase Reporter in the Cyclin D1 Gene Provides a Modified A549 Cell Line to Screen for Selective STAT3 Inhibitors

Targeting the STAT3 signaling pathway in cancer: Role of synthetic and natural inhibitors

Knockdown of STAT3 expression in SKOV3 cells by biodegradable siRNA–PLGA/CSO conjugate micelles

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