Expression of DDX3 Is Directly Modulated by Hypoxia Inducible Factor-1 Alpha in Breast Epithelial Cells

Botlagunta, Mahendran; Krishnamachary, Balaji; Vesuna, Farhad; Winnard, Paul T.; Bol, Guus M.; Patel, Arvind H.; Raman, Venu

doi:10.1371/journal.pone.0017563

Cited by 40 publications

(37 citation statements)

References 46 publications

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“…We also identified ATP dependent RNA helicase (DDX3X) to be up-regulated by hypoxia, which was reported to be directly modulated by HIF-1 α in breast epithelial cells. 45 In addition, we identified several proteins in our study that have not previously been reported to be up-regulated by hypoxia, such as coronin (COR1B), ATP dependent RNA helicase (DDX17), von Hippel-Lindau-binding protein 1 (VBP1), membrane-organizing extension spike protein (MOES), radixin (RADI), and T-complex protein 1 subunit beta (TCPB) (Table S1, Supporting Information). …”

Section: Resultsmentioning

confidence: 83%

MALDI-Mass Spectrometric Imaging Revealing Hypoxia-Driven Lipids and Proteins in a Breast Tumor Model

et al. 2015

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Hypoxic areas are a common feature of rapidly growing malignant tumors and their metastases and are typically spatially heterogeneous. Hypoxia has a strong impact on tumor cell biology and contributes to tumor progression in multiple ways. To date, only a few molecular key players in tumor hypoxia, such as hypoxia-inducible factor-1 (HIF-1), have been discovered. The distribution of biomolecules is frequently heterogeneous in the tumor volume and may be driven by hypoxia and HIF-1α. Understanding the spatially heterogeneous hypoxic response of tumors is critical. Mass spectrometric imaging (MSI) provides a unique way of imaging biomolecular distributions in tissue sections with high spectral and spatial resolution. In this paper, breast tumor xenografts grown from MDA-MB-231-HRE-tdTomato cells, with a red fluorescent tdTomato protein construct under the control of a hypoxia response element (HRE)-containing promoter driven by HIF-1α, were used to detect the spatial distribution of hypoxic regions. We elucidated the 3D spatial relationship between hypoxic regions and the localization of lipids and proteins by using principal component analysis–linear discriminant analysis (PCA-LDA) on 3D rendered MSI volume data from MDA-MB-231-HRE-tdTomato breast tumor xenografts. In this study, we identified hypoxia-regulated proteins active in several distinct pathways such as glucose metabolism, regulation of actin cytoskeleton, protein folding, translation/ribosome, splicesome, the PI3K-Akt signaling pathway, hemoglobin chaperone, protein processing in endoplasmic reticulum, detoxification of reactive oxygen species, aurora B signaling/apoptotic execution phase, the RAS signaling pathway, the FAS signaling pathway/caspase cascade in apoptosis, and telomere stress induced senescence. In parallel, we also identified colocalization of hypoxic regions and various lipid species such as PC(16:0/18:0), PC(16:0/18:1), PC(16:0/18:2), PC(16:1/18:4), PC(18:0/18:1), and PC(18:1/18:1), among others. Our findings shed light on the biomolecular composition of hypoxic tumor regions, which may be responsible for a given tumor's resistance to radiation or chemotherapy.

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Section: Resultsmentioning

confidence: 83%

MALDI-Mass Spectrometric Imaging Revealing Hypoxia-Driven Lipids and Proteins in a Breast Tumor Model

et al. 2015

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“…We have demonstrated that hypoxia inducible factor HIF-1 induced the expression of DDX3 in two different breast cell lines by binding directly or indirectly to the hypoxia-response element (HRE) in the DDX3 proximal promoter [22]. On the other hand a significant down regulation of DDX3 expression is found in hepatocellular carcinoma (HCCs) from hepatitis B virus (HBV)-positive patients, but not from HCV-positive ones, compared to the corresponding non tumor tissues [23].…”

confidence: 99%

“…ATPase activity assay: DDX3 protein was purified from as per the standard protocol [22]. In brief, bacterial cell lysis was passing through Ni-NTA ©2016 agarose resin (In vitrogen) and the protein was purified by affinity chromatographic method.…”

Section: Mtt Assaymentioning

confidence: 99%

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2016

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Abstract:RNA helicase, DDX3 is a multifunctional enzyme and is known to be associated with several diseases like HIV progression, brain and breast cancer. Some of the ring expanded nucleoside compounds such as REN: NZ51, fused di imidazodiazepine ring (RK33), (Z)-3-(5-(3-bromo benzylidene)-4-oxo-2-thioxothiazolidin-3-yl)-N-(2-hydroxy phenyl) propanamide compound (FE15) have been documented to inhibit DDX3 helicase activity. However, synthesis of these drugs is limited to few research groups. Prevalence of literature study, we found that doxorubicin form strong hydrogen bond interactions with crystallized form of DDX3 using in-silico molecular docking approach. To evaluate the biological inhibitory action of doxorubicin, we performed the ATPase activity assay and anti-cancer activity using H357 cancer cell lines. Results showed that doxorubicin continually declined the inorganic phosphate (Pi) release and inhibited the ATP hydrolysis by directly interacting with DDX3. Anticancer activity was detected by MTT assay. The half maximal inhibitory concentrations of doxorubicin (IC50) for H357 cancer cell line is 50 µM and also doxorubicin significantly down regulated the expression of DDX3. Taken together, our results demonstrate, that inhibition of DDX3 expression by using doxorubicin can be used as an ideal drug candidate to treat DDX3 associated cancer disorder by interacting with unique amino acid residues (Thr 198) and common amino acid residues (Tyr 200 and Thr 201). Background:RNA helicases mainly found in all eukaryotes and most prokaryotes. These were distinguished from others based on highly conserved four amino-acid residues (Asp (D)-Glu (E)-Ala (A)-Asp/His (D/H)) at N-terminal region and helicase domain at C-terminal end [1,2]. This amino acid motif has been found in more than 500 proteins and has shown to represent up to 2% of the open reading frames of a genome and may share overlapping or distinct biological cellular functions [3]. These proteins have shown to associate with several aspects of energydependent RNA metabolism including translation, ribosome biogenesis, and pre-mRNA splicing [4, 5]. Unlike DEAH box genes, DEAD box genes have been identified on all human chromosomes except 15, 18, and 21. This highlights the importance that DEAD box RNA helicases in the physiological management of the cell and its survival. Among several DEAD box RNA helicases, DDX3 is located on X-chromosome at Xp11.3-p11.23 [6]. It has a functional homologue on the Y chromosome, DBY, and this gene product has an activity that is crucial for normal spermatogenesis [7-9]. It has shown to participate in human folliculogenesis and its deletion has shown to represent an important genetic cause of primary amenorrhea or impairment of female fertility [10].

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“…[10][11][12][13][14] We have also reported that hypoxia inducible factor (HIF-1) induce the expression of DDX3 in two different breast cell lines by binding directly or indirectly to the hypoxia-response element (HRE) in the DDX3 proximal promoter. 15 On the other hand, a significant down regulation of DDX3 expression is found in hepatocellular carcinoma (HCCs) from the hepatitis B virus (HBV) positive patients, but not from HCV positive in comparison to the corresponding non tumor tissues. 16 In the hepatocellular carcinoma model, DDX3 was found to act as a tumor suppressor by activating the expression of cyclin dependent kinase inhibitor p21 cip1 .…”

Section: Introductionmentioning

confidence: 99%

In vitro Anti-Cancer Activity of Rosuvastatin and Ketorolac Nanoformulations against DDX3

Bheemanapally¹,

Thimmaraju²,

Kasagoni³

et al. 2017

JYP

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INTRODUCTIONHuman DDX3 gene encodes a polypeptide of 662 amino acids and this protein plays an important role in several aspects of RNA metabolism. 1,2 This protein has two clear distinguishable domains comprised of N-terminal DEAD box domain 1 (211-403 residues) and C-terminal helicase domain 2 (411-575 residues). Both domains displayed RecA-like folds comprising a central β-sheet flanked by α-helices connected by a noncanonical linker of 11 amino acids.3 Expression of DDX3 was detected in several tissues such as testis, colon, lung, liver, skeletal muscle, and kidney 2,4 indicating the universal role of DDX3 in cellular homeostasis. Moreover, elevated expression of DDX3 was found in a highly aggressive metastatic breast cancer cell line, MDA-MB-231, as compared with non-metastatic MCF-7 cells, which show its potential role in aggressive breast cancers and associated metastasis. 5,6 We have previously reported that overexpression of DDX3 in immortalized non-turmorigenic MCF10A cells promoted neoplastic transformation as indicated by down regulation of a cell adhesion molecule, E-cadherin.5 Down-regulation of E-cadherin is a common feature of a variety of metastatic epithelial tumors, including those of the lung, breast and prostate cancer.5,7-9 Hypoxic regions of solid tumors were considered to be the primary sites for the generation of the metastatic phenotype and have been demonstrated to be chemo and radio-resistant. [10][11][12][13][14] We have also reported that hypoxia inducible factor (HIF-1) induce the expression of DDX3 in two different breast cell lines by binding directly or indirectly to the hypoxia-response element (HRE) in the DDX3 proximal promoter. 15 On the other hand, a significant down regulation of DDX3 expression is found in hepatocellular carcinoma (HCCs) from the hepatitis B virus (HBV) positive patients, but not from HCV positive in comparison to the corresponding non tumor tissues. 16 In the hepatocellular carcinoma model, DDX3 was found to act as a tumor suppressor by activating the expression of cyclin dependent kinase inhibitor p21 cip1 . 17 Besides cancer, induced expression of DDX3 was also found in HIV-1 infected cells. 18,19 Overall, it suggests that DDX3 is a multifunctional protein and it plays a specific role in regulatory mechanisms and signaling pathways. Apart from embryonic development, this gene is also involved in multiple diseases like HIV, Neuro-degenerative diseases, hepatocellular carcinoma and brain and breast cancer. Several Synthetic compounds have been discovered to inhibit the function of DDX3 by blocking the function of helicase activity. [20][21][22][23][24] Synthetic drugs are those substances that are produced entirely from chemical reactions in a laboratory. Synthetic drugs most often have shown to suppress the immune system by paralyzing the bone marrow. 25,26 Moreover, it takes an average of about 8 to 12 years from the time a cancer drug enters into clinical trials from the research lab.27,28 Therefore, we focused to identify FDA approved drugs against D...

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Expression of DDX3 Is Directly Modulated by Hypoxia Inducible Factor-1 Alpha in Breast Epithelial Cells

Cited by 40 publications

References 46 publications

MALDI-Mass Spectrometric Imaging Revealing Hypoxia-Driven Lipids and Proteins in a Breast Tumor Model

MALDI-Mass Spectrometric Imaging Revealing Hypoxia-Driven Lipids and Proteins in a Breast Tumor Model

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In vitro Anti-Cancer Activity of Rosuvastatin and Ketorolac Nanoformulations against DDX3

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