26S Proteasome Activity Is Down-Regulated in Lung Cancer Stem-Like Cells Propagated In Vitro

Curcumin, the active ingredient in Curcuma longa, has been in medicinal use since ancient times. However, the therapeutic targets and signaling cascades modulated by curcumin have been enigmatic despite extensive research. Here we identify dual-specificity tyrosine-regulated kinase 2 (DYRK2), a positive regulator of the 26S proteasome, as a direct target of curcumin. Curcumin occupies the ATP-binding pocket of DYRK2 in the cocrystal structure, and it potently and specifically inhibits DYRK2 over 139 other kinases tested in vitro. As a result, curcumin diminishes DYRK2-mediated 26S proteasome phosphorylation in cells, leading to reduced proteasome activity and impaired cell proliferation. Interestingly, curcumin synergizes with the therapeutic proteasome inhibitor carfilzomib to induce apoptosis in a variety of proteasome-addicted cancer cells, while this drug combination exhibits modest to no cytotoxicity to noncancerous cells. In a breast cancer xenograft model, curcumin treatment significantly reduces tumor burden in immunocompromised mice, showing a similar antitumor effect as CRISPR/Cas9-mediated DYRK2 depletion. These results reveal an unexpected role of curcumin in DYRK2-proteasome inhibition and provide a proof-of-concept that pharmacological manipulation of proteasome regulators may offer new opportunities for anticancer treatment.

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Combined Effect of Bortezomib and Menadione Sodium Bisulfite on Proteasomes of Tumor Cells: The Dramatic Decrease of Bortezomib Toxicity in a Preclinical Trial

Cancers ◽

10.3390/cancers10100351 ◽

2018 ◽

Vol 10 (10) ◽

pp. 351 ◽

Cited By ~ 6

Author(s):

Tatiana Astakhova ◽

Alexey Morozov ◽

Pavel Erokhov ◽

Maria Mikhailovskaya ◽

Sergey Akopov ◽

...

Keyword(s):

High Efficiency ◽

Sodium Bisulfite ◽

Proteasome Activity ◽

26S Proteasome ◽

Mammary Adenocarcinoma ◽

Antitumor Drugs ◽

Antitumor Effects ◽

Preclinical Trial ◽

Menadione Sodium Bisulfite

Tumor growth is associated with elevated proteasome expression and activity. This makes proteasomes a promising target for antitumor drugs. Current antitumor drugs such as bortezomib that inhibit proteasome activity have significant side effects. The purpose of the present study was to develop effective low-toxic antitumor compositions with combined effects on proteasomes. For compositions, we used bortezomib in amounts four and ten times lower than its clinical dose, and chose menadione sodium bisulfite (MSB) as the second component. MSB is known to promote oxidation of NADH, generate superoxide radicals, and as a result damage proteasome function in cells that ensure the relevance of MSB use for the composition development. The proteasome pool was investigated by the original native gel electrophoresis method, proteasome chymotrypsin-like activity—by Suc-LLVY-AMC-hydrolysis. For the compositions, we detected 10 and 20 μM MSB doses showing stronger proteasome-suppressing and cytotoxic in cellulo effects on malignant cells than on normal ones. MSB indirectly suppressed 26S-proteasome activity in cellulo, but not in vitro. At the same time, MSB together with bortezomib displayed synergetic action on the activity of all proteasome forms in vitro as well as synergetic antitumor effects in cellulo. These findings determine the properties of the developed compositions in vivo: antitumor efficiency, higher (against hepatocellular carcinoma and mammary adenocarcinoma) or comparable to bortezomib (against Lewis lung carcinoma), and drastically reduced toxicity (LD50) relative to bortezomib. Thus, the developed compositions represent a novel generation of bortezomib-based anticancer drugs combining high efficiency, low general toxicity, and a potentially expanded range of target tumors.

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An oral second-generation proteasome inhibitor oprozomib significantly inhibits lung cancer in a p53 independent manner in vitro

Acta Biochimica et Biophysica Sinica ◽

10.1093/abbs/gmz093 ◽

2019 ◽

Vol 51 (10) ◽

pp. 1034-1040 ◽

Cited By ~ 1

Author(s):

Hongge Zhu ◽

Tianhai Wang ◽

Zhou Xin ◽

Yiyi Zhan ◽

Guoming Gu ◽

...

Keyword(s):

Lung Cancer ◽

Cancer Cells ◽

Anticancer Agents ◽

Proteasome Inhibitor ◽

Second Generation ◽

26S Proteasome ◽

Lung Cancer Cells ◽

Independent Manner ◽

P53 Expression

Abstract The destruction of proteins via the ubiquitin–proteasome system is a multi-step, complex process involving polyubiquitination of substrate proteins, followed by proteolytic degradation by the macromolecular 26S proteasome complex. Inhibitors of the proteasome promote the accumulation of proteins that are deleterious to cell survival and are promising anticancer agents. Oprozomib (OPZ), an oral second-generation proteasome inhibitor, has been shown to inhibit the growth of several cancers in preclinical and clinical trials, including multiple myeloma and head and neck cancers, but its effects on lung cancer has not yet been determined. In this study, we evaluated the inhibitory effects of OPZ on lung cancer cell lines in vitro. The results showed that OPZ significantly suppressed cell proliferation and strongly induced apoptosis in both tested lung cancer cells independent of p53 expression. OPZ was able to cause obvious caspase 3 and PARP cleavages and stabilize p53 and its transcriptional targets p21, PUMA, and Noxa. Moreover, OPZ was capable of sensitizing lung cancer cells to the conventional chemotherapeutic drug cisplatin. Our study provides preclinical data and sheds light on the potential applications of proteasome inhibitor OPZ in lung cancer treatment.

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The Phosphatase SHP-2 Activates HIF-1α in Wounds In Vivo by Inhibition of 26S Proteasome Activity

International Journal of Molecular Sciences ◽

10.3390/ijms20184404 ◽

2019 ◽

Vol 20 (18) ◽

pp. 4404 ◽

Cited By ~ 2

Author(s):

Yvonn Heun ◽

Katharina Grundler Groterhorst ◽

Kristin Pogoda ◽

Bjoern F Kraemer ◽

Alexander Pfeifer ◽

...

Keyword(s):

Tyrosine Phosphatase ◽

Hypoxia Inducible Factor ◽

Proteasome Activity ◽

26S Proteasome ◽

Mitogen Activated Protein Kinase ◽

Hypoxic Conditions ◽

Mitogen Activated Protein ◽

Specific Regulation

Vascular remodeling and angiogenesis are required to improve the perfusion of ischemic tissues. The hypoxic environment, induced by ischemia, is a potent stimulus for hypoxia inducible factor 1α (HIF-1α) upregulation and activation, which induce pro-angiogenic gene expression. We previously showed that the tyrosine phosphatase SHP-2 drives hypoxia mediated HIF-1α upregulation via inhibition of the proteasomal pathway, resulting in revascularization of wounds in vivo. However, it is still unknown if SHP-2 mediates HIF-1α upregulation by affecting 26S proteasome activity and how the proteasome is regulated upon hypoxia. Using a reporter construct containing the oxygen-dependent degradation (ODD) domain of HIF-1α and a fluorogenic proteasome substrate in combination with SHP-2 mutant constructs, we show that SHP-2 inhibits the 26S proteasome activity in endothelial cells under hypoxic conditions in vitro via Src kinase/p38 mitogen-activated protein kinase (MAPK) signalling. Moreover, the simultaneous expression of constitutively active SHP-2 (E76A) and inactive SHP-2 (CS) in separate hypoxic wounds in the mice dorsal skin fold chamber by localized magnetic nanoparticle-assisted lentiviral transduction showed specific regulation of proteasome activity in vivo. Thus, we identified a new additional mechanism of SHP-2 mediated HIF-1α upregulation and proteasome activity, being functionally important for revascularization of wounds in vivo. SHP-2 may therefore constitute a potential novel therapeutic target for the induction of angiogenesis in ischemic vascular disease.

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Bufalin derivative BF211 inhibits proteasome activity in human lung cancer cells in vitro by inhibiting β1 subunit expression and disrupting proteasome assembly

Acta Pharmacologica Sinica ◽

10.1038/aps.2016.30 ◽

2016 ◽

Vol 37 (7) ◽

pp. 908-918 ◽

Cited By ~ 13

Author(s):

Peng Sun ◽

Li-xing Feng ◽

Dong-mei Zhang ◽

Miao Liu ◽

Wang Liu ◽

...

Keyword(s):

Lung Cancer ◽

Cancer Cells ◽

Human Lung ◽

Proteasome Activity ◽

Lung Cancer Cells ◽

Human Lung Cancer ◽

Human Lung Cancer Cells ◽

Subunit Expression

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HSP70-driven molecular response to the proteasome machinery inhibition is a vulnerability in cancer

10.1101/2021.11.16.468807 ◽

2021 ◽

Author(s):

Magdalena Oron ◽

Marcin Grochowski ◽

Akanksha Jaiswar ◽

Magdalena Nowak-Niezgoda ◽

Malgorzata Kolos ◽

...

Keyword(s):

Multiple Myeloma ◽

Cancer Cells ◽

Proteasome Inhibition ◽

Cell Types ◽

Proteasome Activity ◽

26S Proteasome ◽

Molecular Response ◽

Compensatory Mechanisms ◽

Compensatory Response

Human neoplasias are often addicted to the cellular proteasome machinery. This has led to the development of bortezomib and carfilzomib proteasome inhibitors, approved for the treatment of multiple myeloma. Cancers, however, were found resistant to the proteasome inhibition in clinical trials, suggesting effective, cancer-specific compensatory responses. Here we employed global proteomics to determine contributions of compensatory mechanisms upon the proteasome inhibition with carfilzomib - in the cells of multiple myeloma, normal fibroblasts, and cancers of lung, colon, and pancreas. A pathway-oriented siRNA screen based on proteomics results showed that molecular chaperones, autophagy- and endocytosis-related proteins are cancer-specific vulnerabilities in combination with carfilzomib. HSP70 family chaperones HSPA1A/B were the most universal proteasome inhibition responders in the proteomes of all the studied cell types and HSPA1A/B inhibition most specifically sensitized cancer cells to carfilzomib in cell lines, patient-derived organoids and mouse xenografts. Overlap of proteomics with RNA-seq data showed that the proteasome inhibition-dependent HSPA1A/B induction in cancer cells is mainly transcription-driven and HSF1/2-dependent. Consequently, we found that a high level of HSPA1A/B mRNA is associated with a low proteasome activity in cancer patient tissues and is a risk factor in cancer patients with the low level of expression of the proteasome. Functionally, the HSPA1A/B induction does not affect a proteasome expression bounce-back upon the carfilzomib treatment, while it supports other mechanisms of the proteasome inhibition response - autophagy, unfolded protein response, and directly the 26S proteasome activity. We found that the 26S proteasome is chaperoned and protected from the inhibition with carfilzomib by HSPA1A/B assisted by DNAJB1 co-chaperone in cancer cells and using purified protein system in vitro. Thus, we define HSPA1A/B as a central player in the cellular compensatory response to the decreased proteasome activity, and the sensitive target in cancer cells with the inhibited proteasome.

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Role of 4-O Galloylchlorogenic Acid in Lung Cancer- An Insilico Approach

International Journal of Pharmaceutical and Clinical Research ◽

10.25258/ijpcr.v9i5.8595 ◽

2017 ◽

Vol 9 (5) ◽

Author(s):

Jaynthy C. ◽

N. Premjanu ◽

Abhinav Srivastava

Keyword(s):

Lung Cancer ◽

Cell Proliferation ◽

In Silico ◽

Computational Study ◽

Regulation Mechanism ◽

Down Regulation ◽

Fruit Extract ◽

In Vitro Techniques ◽

Discovery Studio

Cancer is a major disease with millions of patients diagnosed each year with high mortality around the world. Various studies are still going on to study the further mechanisms and pathways of the cancer cell proliferation. Fucosylation is one of the most important oligosaccharide modifications involved in cancer and inflammation. In cancer development increased core fucosylation by FUT8 play an important role in cell proliferation. Down regulation of FUT8 expression may help cure lung cancer. Therefore the computational study based on the down regulation mechanism of FUT8 was mechanised. Sapota fruit extract, containing 4-Ogalloylchlorogenic acid was used as the inhibitor against FUT-8 as target and docking was performed using in-silico tool, Accelrys Discovery Studio. There were several conformations of the docked result, and conformation 1 showed 80% dock score between the ligand and the target. Further the amino acids of the inhibitor involved in docking were studied using another tool, Ligplot. Thus, in-silico analysis based on drug designing parameters shows that the fruit extract can be studied further using in-vitro techniques to know its pharmacokinetics.

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USP9X-mediated KDM4C deubiquitination promotes lung cancer radioresistance by epigenetically inducing TGF-β2 transcription

Cell Death and Differentiation ◽

10.1038/s41418-021-00740-z ◽

2021 ◽

Author(s):

Xiaohua Jie ◽

William Pat Fong ◽

Rui Zhou ◽

Ye Zhao ◽

Yingchao Zhao ◽

...

Keyword(s):

Lung Cancer ◽

Affinity Purification ◽

Smad Signaling ◽

Lung Cancer Patients ◽

Lysine Specific Demethylase ◽

Underlying Mechanisms ◽

H3k9me3 Level ◽

Critical Binding

AbstractRadioresistance is regarded as the main barrier to effective radiotherapy in lung cancer. However, the underlying mechanisms of radioresistance remain elusive. Here, we show that lysine-specific demethylase 4C (KDM4C) is overexpressed and correlated with poor prognosis in lung cancer patients. We provide evidence that genetical or pharmacological inhibition of KDM4C impairs tumorigenesis and radioresistance in lung cancer in vitro and in vivo. Moreover, we uncover that KDM4C upregulates TGF-β2 expression by directly reducing H3K9me3 level at the TGF-β2 promoter and then activates Smad/ATM/Chk2 signaling to confer radioresistance in lung cancer. Using tandem affinity purification technology, we further identify deubiquitinase USP9X as a critical binding partner that deubiquitinates and stabilizes KDM4C. More importantly, depletion of USP9X impairs TGF-β2/Smad signaling and radioresistance by destabilizing KDM4C in lung cancer cells. Thus, our findings demonstrate that USP9X-mediated KDM4C deubiquitination activates TGF-β2/Smad signaling to promote radioresistance, suggesting that targeting KDM4C may be a promising radiosensitization strategy in the treatment of lung cancer.

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MSCs-engineered biomimetic PMAA nanomedicines for multiple bioimaging-guided and photothermal-enhanced radiotherapy of NSCLC

Journal of Nanobiotechnology ◽

10.1186/s12951-021-00823-6 ◽

2021 ◽

Vol 19 (1) ◽

Author(s):

Yipengchen Yin ◽

Yongjing Li ◽

Sheng Wang ◽

Ziliang Dong ◽

Chao Liang ◽

...

Keyword(s):

Lung Cancer ◽

Cell Membranes ◽

Imaging System ◽

Fluorescence Signal ◽

Bimodal Imaging ◽

Red Blood Cell Membranes ◽

Magnetic Resonance Imaging Mri ◽

Tumor Accumulation

Abstract Background The recently developed biomimetic strategy is one of the mostly effective strategies for improving the theranostic efficacy of diverse nanomedicines, because nanoparticles coated with cell membranes can disguise as “self”, evade the surveillance of the immune system, and accumulate to the tumor sites actively. Results Herein, we utilized mesenchymal stem cell memabranes (MSCs) to coat polymethacrylic acid (PMAA) nanoparticles loaded with Fe(III) and cypate—an derivative of indocyanine green to fabricate Cyp-PMAA-Fe@MSCs, which featured high stability, desirable tumor-accumulation and intriguing photothermal conversion efficiency both in vitro and in vivo for the treatment of lung cancer. After intravenous administration of Cyp-PMAA-Fe@MSCs and Cyp-PMAA-Fe@RBCs (RBCs, red blood cell membranes) separately into tumor-bearing mice, the fluorescence signal in the MSCs group was 21% stronger than that in the RBCs group at the tumor sites in an in vivo fluorescence imaging system. Correspondingly, the T1-weighted magnetic resonance imaging (MRI) signal at the tumor site decreased 30% after intravenous injection of Cyp-PMAA-Fe@MSCs. Importantly, the constructed Cyp-PMAA-Fe@MSCs exhibited strong photothermal hyperthermia effect both in vitro and in vivo when exposed to 808 nm laser irradiation, thus it could be used for photothermal therapy. Furthermore, tumors on mice treated with phototermal therapy and radiotherapy shrank 32% more than those treated with only radiotherapy. Conclusions These results proved that Cyp-PMAA-Fe@MSCs could realize fluorescence/MRI bimodal imaging, while be used in phototermal-therapy-enhanced radiotherapy, providing desirable nanoplatforms for tumor diagnosis and precise treatment of non-small cell lung cancer.

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