All-optical Mechanobiology Interrogation of Yes-associated Protein in Human Cancer and Normal Cells using a Multi-functional System

Cancer cells, relative to normal cells, demonstrate increased sensitivity to glucose-deprivation-induced cytotoxicity. To determine whether oxidative stress mediated by O2•− and hydroperoxides contributed to the differential susceptibility of human epithelial cancer cells to glucose deprivation, the oxidation of DHE (dihydroethidine; for O2•−) and CDCFH2 [5- (and 6-)carboxy-2′,7′-dichlorodihydrofluorescein diacetate; for hydroperoxides] was measured in human colon and breast cancer cells (HT29, HCT116, SW480 and MB231) and compared with that in normal human cells [FHC cells, 33Co cells and HMECs (human mammary epithelial cells)]. Cancer cells showed significant increases in DHE (2–20-fold) and CDCFH2 (1.8–10-fold) oxidation, relative to normal cells, that were more pronounced in the presence of the mitochondrial electron-transport-chain blocker, antimycin A. Furthermore, HCT116 and MB231 cells were more susceptible to glucose-deprivation-induced cytotoxicity and oxidative stress, relative to 33Co cells and HMECs. HT29 cells were also more susceptible to 2DG (2-deoxyglucose)-induced cytotoxicity, relative to FHC cells. Overexpression of manganese SOD (superoxide dismutase) and mitochondrially targeted catalase significantly protected HCT116 and MB231 cells from glucose-deprivation-induced cytotoxicity and oxidative stress and also protected HT29 cells from 2DG-induced cytotoxicity. These results show that cancer cells (relative to normal cells) demonstrate increased steady-state levels of ROS (reactive oxygen species; i.e. O2•− and H2O2) that contribute to differential susceptibility to glucose-deprivation-induced cytotoxicity and oxidative stress. These studies support the hypotheses that cancer cells increase glucose metabolism to compensate for excess metabolic production of ROS and that inhibition of glucose and hydroperoxide metabolism may provide a biochemical target for selectively enhancing cytotoxicity and oxidative stress in human cancer cells.

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Identification of Parasporin Genes in Vietnamese Isolates of Bacillus thuringiensis

Zeitschrift für Naturforschung C ◽

10.1515/znc-2008-1-225 ◽

2008 ◽

Vol 63 (1-2) ◽

pp. 139-143 ◽

Cited By ~ 7

Author(s):

Koichi Yasutake ◽

Akiko Uemori ◽

Ngo D. Binh ◽

Eiichi Mizuki ◽

Michio Ohba

Keyword(s):

Bacillus Thuringiensis ◽

Cancer Cells ◽

Human Cancer ◽

Nucleotide Sequence Analysis ◽

Hep G2 ◽

Normal Cells ◽

Proteolytic Activation ◽

Human Cancer Cells ◽

Genes Encoding

Four genes encoding parasporins, cytotoxins preferentially killing human cancer cells in vitro, were isolated from four Vietnamese strains of Bacillus thuringiensis. Nucleotide sequence analysis revealed that: (1) three genes fall into the two known classes, ps1Aa and ps1Ab, and (2) another one belongs to ps1Ac, a novel gene class established in this study. Upon proteolytic activation, parasporal protein of the organism with ps1Ac exhibited strong cytocidal activity against human cancer cells, HeLa and Hep G2, but not to non-cancer normal cells, UtSMC and HC.

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LN-6: a monoclonal antibody to vimentin expressed in non-hematopoietic mesenchymal cells and derived tumors and reactive in B5-fixed, paraffin-embedded tissues.

Journal of Histochemistry & Cytochemistry ◽

10.1177/37.9.2671152 ◽

1989 ◽

Vol 37 (9) ◽

pp. 1363-1370 ◽

Cited By ~ 4

Author(s):

E Stathopoulos ◽

G S Naeve ◽

C R Taylor ◽

A L Epstein

Keyword(s):

Monoclonal Antibody ◽

Human Cancer ◽

Wide Spectrum ◽

Mesenchymal Cells ◽

Lymphoid Cells ◽

Normal Cells ◽

Immunohistochemical Diagnosis ◽

Pathological Tissues

We generated a monoclonal antibody (MAb), designated LN-6, directed against human vimentin, which retains its immunoreactivity in B5-fixed, paraffin-embedded tissues. Like other anti-vimentin MAb, LN-6 was found to be reactive with a wide spectrum of human sarcomas and normal cells of mesenchymal derivation. However, unlike other similar reagents, LN-6 was unreactive with normal and malignant human lymphoid cells and therefore displays a more restricted immunoreactivity. Because of its ability to stain routinely processed pathological tissues and its marked reactivity with human sarcomas, LN-6 is a unique reagent for the immunohistochemical diagnosis of human cancer.

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Revisiting Szent-Györgyi's Cancer Hypothesis

10.21203/rs.3.rs-754303/v1 ◽

2021 ◽

Author(s):

Morris Alfred Johnson ◽

Michael Smits

Keyword(s):

Cell Lines ◽

Strong Interaction ◽

Human Cancer ◽

Glyoxalase I ◽

Human Cancer Cell ◽

Normal Cells ◽

Human Cancer Cell Lines ◽

Mouse Lymphoma Cells ◽

Near Term ◽

Mouse Lymphoma

Abstract Rationale for this communication • -The promine/retine hypothesis on the control of cancer never reached a definite conclusion.• -The chemical natures of promine and retine remain unsettled though usually assumed to be glyoxalases and methylglyoxal.• -Many years ago we published some data that indicated the hypothesis may be operating in plants in which glyoxalase I may exist in normal cells in an inhibited state rather than compartmentalized as in early versions of the hypothesis.• -Manju Ray in India has published many papers claiming that methylglyoxal can be used to successfully treat cancer in humans.• -We present here previously unpublished data that shows depriving glyoxalase I of GSH allows methylglyoxal to kill mouse lymphoma cells. During treatment of two human cancer cell lines, killing of one line was enhanced by blocking thioredoxin as well as GSH.It is hoped that what is conveyed here may reignite interest in the near term. Nuclear methodology and statistics can be found in Figure 1. The data show a strong interaction between the hypothesis and thiols. It is concluded that the hypothesis has yet to be thoroughly investigated.

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Activation of PTHrP-cAMP-CREB1 signaling following p53 loss is essential for osteosarcoma initiation and maintenance

eLife ◽

10.7554/elife.13446 ◽

2016 ◽

Vol 5 ◽

Cited By ~ 22

Author(s):

Mannu K Walia ◽

Patricia MW Ho ◽

Scott Taylor ◽

Alvin JM Ng ◽

Ankita Gupte ◽

...

Keyword(s):

Human Cancer ◽

Point Mutations ◽

Therapeutic Targets ◽

Complete Loss ◽

Osteoblastic Cells ◽

Normal Cells ◽

P53 Pathway ◽

Common Cancer ◽

Frequent Event ◽

Camp Levels

Mutations in the P53 pathway are a hallmark of human cancer. The identification of pathways upon which p53-deficient cells depend could reveal therapeutic targets that may spare normal cells with intact p53. In contrast to P53 point mutations in other cancer, complete loss of P53 is a frequent event in osteosarcoma (OS), the most common cancer of bone. The consequences of p53 loss for osteoblastic cells and OS development are poorly understood. Here we use murine OS models to demonstrate that elevated Pthlh (Pthrp), cAMP levels and signalling via CREB1 are characteristic of both p53-deficient osteoblasts and OS. Normal osteoblasts survive depletion of both PTHrP and CREB1. In contrast, p53-deficient osteoblasts and OS depend upon continuous activation of this pathway and undergo proliferation arrest and apoptosis in the absence of PTHrP or CREB1. Our results identify the PTHrP-cAMP-CREB1 axis as an attractive pathway for therapeutic inhibition in OS.

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A cancer-unique glycan: de-N-acetyl polysialic acid (dPSA) linked to cell surface nucleolin depends on re-expression of the fetal polysialyltransferase ST8SIA2 gene

Journal of Experimental & Clinical Cancer Research ◽

10.1186/s13046-021-02099-y ◽

2021 ◽

Vol 40 (1) ◽

Author(s):

Gregory R. Moe ◽

Lindsay M. Steirer ◽

Joshua A. Lee ◽

Adarsha Shivakumar ◽

Alejandro D. Bolanos

Keyword(s):

Cell Surface ◽

Cell Lines ◽

Cancer Cell ◽

Fetal Development ◽

Blood Cells ◽

Human Cancer ◽

Polysialic Acid ◽

Human Cancer Cell ◽

Normal Cells ◽

Human Cancer Cell Lines

Abstract Background Polysialic acid (polySia) modifies six cell surface proteins in humans mainly during fetal development and some blood cells in adults. Two genes in humans, ST8SIA2 and ST8SIA4, code for polysialyltransferases that synthesize polySia. ST8SIA2 is highly expressed during fetal development and in cancer but not in adult normal human cells. ST8SIA4 is expressed in fetal and adult brain, spleen, thymus, and peripheral blood leukocytes and in cancer. We identified a derivative of polySia containing de-N-acetyl neuraminic acid residues (dPSA), which is expressed on the cell surface of human cancer cell lines and tumors but not normal cells. Methods dPSA-modified proteins in several human cancer cell lines and normal blood cells were identified using co-immunoprecipitation with anti-dPSA antibodies, mass spectroscopy and Western blot. RNAi and CRISPR were used to knockdown and knockout, respectively, the polysialyltransferase genes in human melanoma SK-MEL-28 and neuroblastoma CHP-134 cell lines, respectively, to determine the effect on production of cell surface dPSA measured by flow cytometry and fluorescence microscopy. Results We found that dPSA is linked to or associated with nucleolin, a nuclear protein reported to be on the cell surface of cancer but not normal cells. Knocking down expression of ST8SIA2 with RNAi or knocking out each gene individually and in combination using CRISPR showed that cell surface dPSA depended on expression of ST8SIA2. Conclusions The presence of dPSA specifically in a broad range of human cancers but not human adult normal cells offers novel possibilities for diagnosis, prevention and treatment targeting the dPSA antigen that appears to be cancer-specific, consistent across not only human cancers but also species, and may be an unrecognized mechanism of immune shielding.

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