Copper deprivation enhances the chemosensitivity of pancreatic cancer to rapamycin by mTORC1/2 inhibition.

Cuproplasia, or copper-dependent cell proliferation, has been observed in varieties of solid tumors along with aberrant copper homeostasis. Several studies reported good response of patients to copper chelator assisted neoadjuvant chemotherapy, however, the internal target molecules are still undetermined. Unravel copper-associated tumor signaling would be valuable to forge new links to translate biology of copper into clinical cancer therapies. We evaluated the significance of high-affinity copper transporter-1 (CTR1) by bioinformatic analysis, and in 19 pairs of clinical specimens. Then, with the help of gene interference and chelating agent, enriched signaling pathways were identified by KEGG analysis and immunoblotting. Accompanying biological capability of pancreatic carcinoma-associated proliferation, cell cycle, apoptosis, and angiogenesis were investigated. Furthermore, a combination of mTOR inhibitor and CTR1 suppressor has been assessed in xenografted tumor mouse models. Hyperactive CTR1 was investigated in pancreatic cancer tissues and proven to as the key point of cancer copper homeostasis. Intracellular copper deprivation induced by CTR1 gene knock-down or systematic copper chelation by tetrathiomolybdate suppressed proliferation and angiogenesis of pancreatic cancer cell. PI3K/AKT/mTOR signaling pathway was suppressed by inhibiting the activation of p70(S6)K and p-AKT, and finally inhibited mTORC1 and mTORC2 after copper deprivation. Additionally, CTR1 gene silencing successfully improved the anti-cancer effect of mTOR inhibitor rapamycin. Our study reveals that CTR1 contributes to pancreatic tumorigenesis and progression, by up-regulating the phosphorylation of AKT/mTOR signaling molecules. Recovering copper balance by copper deprivation addresses as promising strategy for improved cancer chemotherapy.

Identification of the effect and mechanism of Yiyi Fuzi Baijiang powder against colorectal cancer using network pharmacology and experimental validation.

Background: Yiyi Fuzi Baijiang powder (YFBP) is a traditional Chinese medicine used to treat colorectal cancer, although its bioactivity and mechanisms of action have not been studied in depth yet. The study intended to identify the potential targets and signaling pathways affected by YFBP during the treatment of colorectal cancer through pharmacological network analysis and to further analyze its chemical compositions and molecular mechanisms of action. Methods: The Traditional Chinese Medicine Systems Pharmacology (TCMSP), Traditional Chinese Medicine Integrated Database (TCMID), HitPredict (HIT), and Search Tool for Interactions of Chemicals (STITCH) databases were used to screen the bioactive components and promising targets of YFBP. Targets related to colorectal cancer were retrieved from the GeneCards and Gene Ontology databases. Cytoscape software was used to construct the "herb-active ingredient-target" network. The STRING database was used to construct and analyze protein-protein interactions (PPIs). Afterward, the R packages clusterProfiler and Cytoscape Hub plug-in were used to perform Gene Ontology (GO) functional and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of target genes. The results of the network pharmacological analysis were also experimentally validated. Results: In total, 33 active components and 128 target genes were screened. Among them, 46 target genes were considered potential therapeutic targets that crossed the CRC target genes. The network pharmacology analysis showed that the active components of YFBP were correlated positively with CRC inflammatory target genes such as TLR4, TNF, and IL-6. The inflammation-related signaling pathways affected by the active components included the TNF-α, interleukin-17, and toll-like receptor signaling pathways. The active ingredients of YFBP, such as luteolin, ß-sitosterol, myristic acid, and vanillin, may exert anti-tumor effects by downregulating SMOX expression via anti-inflammatory signaling and regulation of the TLR4/NF-κB signaling pathway. Conclusion: In the present study, the potential active components, potential targets, and key biological pathways involved in the YFBP treatment of CRC were determined, providing a theoretical foundation for further anti-tumor research.

Vascular Calcification in Chronic Kidney Disease: An Update and Perspective.

Chronic kidney disease is a devastating condition resulting from irreversible loss of nephron numbers and function and leading to end-stage renal disease and mineral disorders. Vascular calcification, an ectopic deposition of calcium-phosphate salts in blood vessel walls and heart valves, is an independent risk factor of cardiovascular morbidity and mortality in chronic kidney disease. Moreover, aging and related metabolic disorders are essential risk factors for chronic kidney disease and vascular calcification. Marked progress has been recently made in understanding and treating vascular calcification in chronic kidney disease. However, there is a paucity of systematic reviews summarizing this progress, and investigating unresolved issues is warranted. In this systematic review, we aimed to overview the underlying mechanisms of vascular calcification in chronic kidney diseases and discuss the impact of chronic kidney disease on the pathophysiology of vascular calcification. Additionally, we summarized potential clinical diagnostic biomarkers and therapeutic applications for vascular calcification with chronic kidney disease. This review may offer new insights into the pathogenesis, diagnosis, and therapeutic intervention of vascular calcification.

Effects of ΔNp63 Gene Down-expression on Invasion of Bladder Carcinoma Cells in Vitro.

PURPOSE: This work aims to investigate the effects of ΔNp63 gene down-expression on invasion of bladder carcinoma cells in vitro. MATERIALS AND METHODS: Bladder carcinoma cell lines UM-UC-3 and 5637 were cultured. The expression plasmids encoding ΔNp63 were constructed and transfected into UM-UC-3 and 5637 cells. The migration and adhesion of cells were detected. The expressions of ΔNp63 and invasion-related zonula occludens protein-1 (ZO-1) in cells were determined by real-time polymerase chain reaction (PCR) and western blot analysis. Confocal microscopy was used to observe the location of ZO-1 in cells. RESULTS: Results showed that the down-expression of ΔNp63 reduced the migration of UM-UC-3 and 5637 cells, decreased the heterogeneity adhesion, and increased homogeneous adhesion. After transfection with ΔNp63, the ZO-1 expression in cell membrane and cell cytoplasm was inhibited, also the ZO-1 mRNA and protein levels in cells were significantly decreased. CONCLUSION: This study indicates thatΔNp63 gene down-expression can reduce the invasion of bladder carcinoma cells in vitro.

HOXB2 and FOXC1 synergistically drive the progression of Wilms tumor.

OBJECTIVE: To uncover the expression patterns of HOXB2 and FOXC1 in Wilms tumor samples, and their synergistical regulations on the development of Wilms tumor. METHODS: Expression levels of HOXB2 and FOXC1 in 58 cases of Wilms tumor tissues and paracancerous ones were detected. The influences of HOXB2 and FOXC1 on prognosis in Wilms tumor patients were analyzed. Their regulatory effects on proliferative and migratory abilities in WT-CLS1 and HFWT cells were examined by cell counting kit-8 (CCK-8) and Transwell assay, respectively. The interaction between HOXB2 and FOXC1, and their synergistical regulation on the development of Wilms tumor were finally explored. RESULTS: HOXB2 and FOXC1 were upregulated in Wilms tumor tissues. Higher levels of HOXB2 and FOXC1 indicated higher risks of advanced stage and lymphatic metastasis, as well as worse prognosis in Wilms tumor patients. Knockdown of HOXB2 or FOXC1 weakened proliferative and migratory abilities in WT-CLS1 and HFWT cells, while the opposite trends were observed in those overexpressing HOXB2 or FOXC1. The positive interaction between HOXB2 and FOXC1 was identified, which synergistically drove the malignant development of Wilms tumor. CONCLUSIONS: HOXB2 and FOXC1 are upregulated in Wilms tumor samples, and they are closely linked to tumor staging and lymphatic metastasis in Wilms tumor patients. HOXB2 and FOXC1 synergistically drive the malignant development of Wilms tumor by stimulating proliferative and migratory potentials.

[Clinical study of concealed penis correction surgery based on principle of midline symmetry].

OBJECTIVE: To investigate the effectiveness of concealed penis correction surgery based on the principle of midline symmetry. METHODS: Between January 2016 and September 2018, 18 children with concealed penis were treated with correction surgery based on the principle of midline symmetry. All children were 3-12 years old, with an average age of 8.3 years. Physical examination showed that the penis was short; the penis body could not be exposed or be exposed too limited; the corpus cavernosum developed well. The pressure dressing was removed at 3 days after operation and the urethral tube was removed. The color of the glans, the swelling and congestion of penis and scrotum, and the blood supple of the prepuce flap were observed. RESULTS: The operation time ranged from 47 to 54 minutes, with an average of 50 minutes. All children were followed up 3 months after operation. There was no hemorrhage and necrosis of the glans and no infection or ischemic necrosis of the flap. All patients had different degree of prepuce edema at 3 days after operation, 5 patients still had prepuce edema at 2 weeks, and the prepuce edema in all patients subsided at 3 months. All penises were exposed well after midline symmetric anastomosis with no bulky prepuce and scrotum. CONCLUSION: The correction surgery based on the principle of midline symmetry can be used to correct the appearance of the concealed penis effectively.

The PI3K/AKT axis modulates AATF activity in Wilms' tumor cells.

Previous studies have reported excessive expression of apoptosis-antagonizing transcription factor (AATF) in various tumors, where it reinforces the generation and development of cancers and is linked to the clinical outcome. Nevertheless, the expression and influence of AATF in Wilms' tumor (WT) is largely unknown. Here, we discovered that AATF expression was markedly increased in WT tissues as compared to the surrounding normal tissues. Elevated levels of AATF expression were related to tumor relapse and pulmonary metastasis, congruent with it being a predictor of clinical outcome in people suffering from WT. Proliferation, invasion, and migration of WT cells were suppressed by knockdown of AATF and promoted by AATF overexpression in vitro. Furthermore, the tumor generation capability of WT cells noticeably decreased after knockout of AATF in vivo. The phosphoinositide-3-kinase (PI3K)/AKT pathway modulated the activity of AATF in WT. The findings of our study indicate that AATF expression is increased in WT and can serve as a predictor of clinical outcome; in addition, it may enhance the development of WT via the PI3K/AKT axis and may be a promising marker for WT diagnosis and therapy.

IL-8 and IP-10 expression from human bronchial epithelial cells BEAS-2B are promoted by Streptococcus pneumoniae endopeptidase O (PepO).

BACKGROUND: The bronchial epithelium serves as the first defendant line of host against respiratory inhaled pathogens, mainly through releasing chemokines (e.g. interleukin-8 (IL-8), interferon-induced protein 10 (IP-10) etc.) responsible for neutrophil or lymphocyte recruitment to promote the clearance of inhaled pathogens including Streptococcus pneumoniae (S. pneumoniae). Previous studies have shown that IL-8 expression is induced by pneumococcal virulence factors (e.g. pneumolysin, peptidoglycan-polysaccharides, pneumococcal surface protein A (PspA) etc.), which contributes to the pathogenesis of pneumonia. Whether other pneumococcal virulence factors are involved in inducing chemokines expression in epithelium is still unknown. RESULTS: We studied the effect of PepO, a widely expressed and newly discovered pneumococcal virulence protein, on the release of proinflammatory cytokines, IL-8 and IP-10, from human bronchial epithelial cell line BEAS-2B and identified the relevant signaling pathways. Incubation of BEAS-2B with PepO resulted in increased synthesis and release of IL-8 and IP-10 in a dose and time independent manner. We also detected the increased and sustained expression of TLR2 and TLR4 transcripts in BEAS-2B stimulated by PepO. PepO activation leaded to the phosphorylation of MAPKs, Akt and p65. Pharmacologic inhibitors of MAPKs, PI3K and IκB-α phosphorylation attenuated IL-8 release, while IP-10 production was just suppressed by inhibitors of IκB-α phosphorylation, PI3K and P38 MAPK. CONCLUSION: These results suggest that PepO enhances IL-8 and IP-10 production in BEAS-2B in a MAPKs-PI3K/Akt-p65 dependent manner, which may play critical roles in the pathogenesis of pneumonia.

ΔNp63 promotes UM­UC­3 cell invasiveness and migration through claudin­1 in vitro.

The p63 gene, a member of the p53 gene family, has two different promoter usage­generating proteins that contain or lack (ΔN) an NH2­terminus. Although p53 and p63 have high sequence and structural similarities, the molecules differ in function and expression profiles. p63 is critical for the development of epithelial organs or tissues, including the epidermis and other squamous epithelia, as well as the salivary, lachrymal, mammary and prostate glands and the urothelium. In addition, p63 is essential for the proliferative potential of stem cells in the epidermis. In contrast to p53, the role of ΔNp63 in tumors remains unclear and complex. Our previous study demonstrated that ΔNp63 is overexpressed in human bladder carcinoma tissues. The mechanism by which ΔNp63 promotes tumor cell development, including adhesion, proliferation and polarity, is unknown. Data demonstrate that ΔNp63 induces the invasiveness of cancer cells through specific downstream genes and the mechanism is associated with cell junctions. Claudin­1 is an important p63 target gene for normal skin development. Claudin­1, as a connexin, functions in a similar manner to other connexins to affect important events during cancer cell development. In the present study, ΔNp63 gene expression in bladder tumor tissues was found to be significantly higher than that in normal tissue, indicating that ΔNp63 is localized to the nucleus. In addition, ΔNp63 silencing decreased invasion and metastasis in UM­UC­3 cells and reduced claudin­1 expression.