shikonin and Pancreatic-Neoplasms

Shikonin is the main component of root extracts from the Chinese herbal medicine

Topics: Apoptosis; Cell Line, Tumor; Cell Proliferation; Humans; Molecular Docking Simulation; Naphthoquinones; p21-Activated Kinases; Pancreatic Neoplasms

Pancreatic cancer (PC) is a highly fatal malignancy with few effective therapies currently available. Recent studies have shown that PD-L1 inhibitors could be potential therapeutic targets for the treatment of PC. The present study aims to investigate the effect of Shikonin on immune evasion in PC with the involvement of the PD-L1 degradation.. Initially, the expression patterns of PD-L1 and NF-κB in PC were predicted in-silico using the GEPIA database, and were subsequently validated using PC tissues. Thereafter, the correlation of NF-κB with STAT3, CSN5 and PD-L1 was examined. PC cells were treated with Shikonin, NF-κB inhibitor, STAT3 activator, and CSN5 overexpression plasmid to investigate effects on PD-L1 glycosylation and immune evasion in PC. Finally, in vivo tumor formation was induced in C57BL/6J mice, in order to verify the in vitro results.. PD-L1, NF-κB, NF-κB p65, STAT3, and CSN5 were highly expressed in PC samples, and NF-κB was positively correlated with STAT3/CSN5/PD-L1. Inhibition of NF-κB decreased PD-L1 glycosylation and increased PD-L1 degradation, whereas activated STAT3 and overexpressed CSN5 reversed these trends. Shikonin blocked immune evasion in PC, and lowered the expression of PD-L1, NF-κB, NF-κB p65, STAT3 and CSN5 in vivo and in vitro.. The findings indicated Shikonin inhibited immune evasion in PC by inhibiting PD-L1 glycosylation and activating the NF-κB/STAT3 and NF-κB/CSN5 signaling pathways. These effects of Shikonin on PC cells may bear important potential therapeutic implications for the treatment of PC.

Topics: Animals; Antineoplastic Agents; B7-H1 Antigen; Biomarkers, Tumor; Blotting, Western; Cell Line, Tumor; COP9 Signalosome Complex; Flow Cytometry; Humans; Immunohistochemistry; Intracellular Signaling Peptides and Proteins; Male; Mice; Mice, Inbred C57BL; Naphthoquinones; NF-kappa B; Pancreatic Neoplasms; Peptide Hydrolases; Signal Transduction; STAT3 Transcription Factor; Tumor Escape; Xenograft Model Antitumor Assays

Pancreatic ductal adenocarcinoma (PDAC) has poor survival and treatment options. PDAC cells shift their metabolism towards glycolysis, which fuels the plasma membrane calcium pump (PMCA), thereby preventing Ca. PDAC cell growth, migration and death were assessed by using sulforhodamine-B/tetrazolium-based assays, gap closure assay and poly-ADP ribose polymerase (PARP1) cleavage, respectively. Cellular ATP and metabolism were assessed using luciferase/fluorescent-based assays and the Seahorse XFe96 analyzer, respectively. Cell surface biotinylation identified membrane-associated proteins. Fura-2 imaging was used to assess cytosolic Ca. The PKM2 inhibitor (shikonin) reduced PDAC cell proliferation, cell migration and induced cell death. This was due to inhibition of glycolysis, ATP depletion, inhibition of PMCA and cytotoxic Ca. Cutting off the PKM2-derived ATP supply to the PMCA represents a novel therapeutic strategy for the treatment of PDAC.

Topics: Adenosine Triphosphate; Calcium; Carcinoma, Pancreatic Ductal; Carrier Proteins; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Cytosol; Glycolysis; Humans; Membrane Proteins; Naphthoquinones; Pancreas; Pancreatic Neoplasms; Thyroid Hormone-Binding Proteins; Thyroid Hormones

Although gemcitabine is currently the best chemotherapeutic agent available for the treatment of advanced pancreatic cancer, eventual failure of response is a significant clinical problem. Therefore, novel therapeutic approaches against this disease are highly needed. The aim of this study was to evaluate whether shikonin, a naphthoquinone derivative, has potential in the treatment of pancreatic cancer when used either alone or in combination with gemcitabine. Our in vitro results showed that shikonin inhibited the proliferation of three different human pancreatic cancer cell lines and potentiated the cytotoxic effect of gemcitabine, which correlated with the down-regulation of constitutive as well as gemcitabine-induced activation of NF-κB and NF-κB-regulated gene products. Most importantly, using a xenograft model of human pancreatic cancer, we found shikonin alone significantly suppressed tumor growth and argumented the antitumor activity of gemcitabine. These effects also correlated with the down-regulation of NF-κB activity and its target genes, decreased proliferation (PCNA and Ki-67), decreased microvessel density (CD31), and increased apoptosis (TUNEL) in tumor remnants. Collectively, our results suggest that shikonin can suppress the growth of human pancreatic tumors and potentiate the antitumor effects of gemcitabine through the suppression of NF-κB and NF-κB-regulated gene products.

Topics: Animals; Antineoplastic Agents; Apoptosis; Cell Survival; Deoxycytidine; Drug Synergism; Gemcitabine; Humans; Male; Mice; Mice, Nude; Microvessels; Naphthoquinones; Neovascularization, Pathologic; NF-kappa B; Pancreatic Neoplasms; Signal Transduction; Xenograft Model Antitumor Assays