Evaluating Cytoplasmic and Nuclear Levels of Inflammatory Cytokines in Cancer Cells by Western Blotting

Singha, Bipradeb; Persaud, Valerie; Vancurova, Ivana

doi:10.1007/978-1-4939-0928-5_25

Cited by 7 publications

(7 citation statements)

References 23 publications

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“…These data showed that HMGB1 overexpression might be correlated with the development and progression of OS. These observations also support previous conclusions that increased expression HMGB1 is associated with increased cell proliferation, angiogenesis, and metastasis during cancer progression [15,33].…”

Section: Discussionsupporting

confidence: 90%

HMGB1 promotes cellular proliferation and invasion, suppresses cellular apoptosis in osteosarcoma

et al. 2014

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Osteosarcoma is the most common primary malignant bone tumor in children and adolescents. Unfortunately, treatment failures are common due to the metastasis and chemoresistance, but the underlying molecular mechanism remains unclear. Accumulating evidence indicated that the deregulation of DNA-binding protein high-mobility group box 1 (HMGB1) was associated with the development of cancer. This study aimed to explore the expression of HMGB1 in osteosarcoma tissues and its correlation to the clinical pathology of osteosarcoma and to discuss the role of HMGB1 in the development of osteosarcoma. The results from RT-PCR and Western blot showed that the expression rate of HMGB1 messenger RNA (mRNA) and the expression of HMGB1 in the osteosarcoma tissues were significantly higher than those in normal bone tissue (p < 0.05), the expression rate of HMGB1 mRNA and the expression of HMGB1 in the carcinoma tissues with positive lung metastasis were significantly higher than those without lung metastasis (p < 0.05), and with increasing Enneking stage, the expression rate of HMGB1 mRNA and the expression of HMGB1 also increased (p < 0.05). In order to explore the role of HMGB1 in osteosarcoma, the expression of HMGB1 in the human osteosarcoma MG-63 cell line was downregulated by the technique of RNA interference. Western blot results showed that the protein expression of HMGB1 was significantly decreased in the MG-63 cells from HMGB1-siRNA transfection group (p < 0.05), which suggested that HMGB1 was successfully downregulated in the MG-63 cells. Then the changes in proliferation, apoptosis, and invasion of MG-63 cells were examined by MTT test, PI staining, annexin V staining, and transwell chamber assay. Results showed that the abilities of proliferation and invasion were suppressed in HMGB1 knockdown MG-63 cells, and the abilities of apoptosis were enhanced in HMGB1 knockdown MG-63 cells. The expression of cyclin D1, MMP-9 was downregulated in HMGB1 knockdown MG-63 cells, and the expression of caspase-3 was upregulated in HMGB1 knockdown MG-63 cells. Taken together, the overexpression of HMGB1 in osteosarcoma might be related to the tumorigenesis, invasion, and metastasis of osteosarcoma, which might be a potential target for the treatment of osteosarcoma.

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Section: Discussionsupporting

confidence: 90%

HMGB1 promotes cellular proliferation and invasion, suppresses cellular apoptosis in osteosarcoma

et al. 2014

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“…Techniques for subcellular fractionation prior cytokine detection can distinguish nuclear and cytoplasmic localization [ 104 , 105 ]. Most of the cytokines are localized in the cytoplasm of the producing cell before secretion.…”

Section: Cytokine Detection Techniquesmentioning

confidence: 99%

Advances in Proteomic Techniques for Cytokine Analysis: Focus on Melanoma Research

Skalníková

Čížková

Cervenka

et al. 2017

IJMS

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Melanoma is a skin cancer with permanently increasing incidence and resistance to therapies in advanced stages. Reports of spontaneous regression and tumour infiltration with T-lymphocytes makes melanoma candidate for immunotherapies. Cytokines are key factors regulating immune response and intercellular communication in tumour microenvironment. Cytokines may be used in therapy of melanoma to modulate immune response. Cytokines also possess diagnostic and prognostic potential and cytokine production may reflect effects of immunotherapies. The purpose of this review is to give an overview of recent advances in proteomic techniques for the detection and quantification of cytokines in melanoma research. Approaches covered span from mass spectrometry to immunoassays for single molecule detection (ELISA, western blot), multiplex assays (chemiluminescent, bead-based (Luminex) and planar antibody arrays), ultrasensitive techniques (Singulex, Simoa, immuno-PCR, proximity ligation/extension assay, immunomagnetic reduction assay), to analyses of single cells producing cytokines (ELISpot, flow cytometry, mass cytometry and emerging techniques for single cell secretomics). Although this review is focused mainly on cancer and particularly melanoma, the discussed techniques are in general applicable to broad research field of biology and medicine, including stem cells, development, aging, immunology and intercellular communication.

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“…HMGB1 is considered a biomarker for ovarian cancer [ 38 , 39 ] and increased levels of interleukin-8 protein (IL-8) and HMGB1 correlate with poor prognosis in prostate and ovarian cancer cells [ 125 ]. Targeting HMGB1 by RNA interference inhibits ovarian cancer growth and metastasis [ 126 ].…”

Section: The Function Of Hmgb Proteins and Other Redox Sensors Durmentioning

confidence: 99%

High Mobility Group B Proteins, Their Partners, and Other Redox Sensors in Ovarian and Prostate Cancer

Barreiro-Alonso¹,

Lamas-Maceiras²,

Rodrı́guez-Belmonte³

et al. 2015

Oxidative Medicine and Cellular Longevity

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Cancer cells try to avoid the overproduction of reactive oxygen species by metabolic rearrangements. These cells also develop specific strategies to increase ROS resistance and to express the enzymatic activities necessary for ROS detoxification. Oxidative stress produces DNA damage and also induces responses, which could help the cell to restore the initial equilibrium. But if this is not possible, oxidative stress finally activates signals that will lead to cell death. High mobility group B (HMGB) proteins have been previously related to the onset and progressions of cancers of different origins. The protein HMGB1 behaves as a redox sensor and its structural changes, which are conditioned by the oxidative environment, are associated with different functions of the protein. This review describes recent advances in the role of human HMGB proteins and other proteins interacting with them, in cancerous processes related to oxidative stress, with special reference to ovarian and prostate cancer. Their participation in the molecular mechanisms of resistance to cisplatin, a drug commonly used in chemotherapy, is also revised.

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Evaluating Cytoplasmic and Nuclear Levels of Inflammatory Cytokines in Cancer Cells by Western Blotting

Cited by 7 publications

References 23 publications

HMGB1 promotes cellular proliferation and invasion, suppresses cellular apoptosis in osteosarcoma

HMGB1 promotes cellular proliferation and invasion, suppresses cellular apoptosis in osteosarcoma

Advances in Proteomic Techniques for Cytokine Analysis: Focus on Melanoma Research

High Mobility Group B Proteins, Their Partners, and Other Redox Sensors in Ovarian and Prostate Cancer

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