D1 Protein Research Articles

TNFα stimulates the proliferation of immature Sertoli cells by attenuating UPS-degradation of cyclin D1 and leads to the delay of BTB maturation in pubertal rats.

The Sertoli cell that plays a vital role during spermatogenesis is a known target of physiological and pathological factors affecting testicular development. Tumor necrosis factor alpha (TNFα) participates in the blood-testis barrier reconstruction, cell apoptosis, and inflammatory response by recognizing receptors on Sertoli cell. TNFα has also been shown to induce the proliferation of immature Sertoli cell in vitro, yet the mechanism still remains unclarified. This study was designed to investigate the effect of TNFα on blood-testis barrier development during puberty and the underlying mechanisms of TNFα-induced immature Sertoli cell proliferation. Immature male Sprague-Dawley rats of postnatal day 12 were intraperitoneally injected with TNFα. Biotin-labeled method was used to detect permeability of the developing blood-testis barrier after TNFα treatment, and the distribution of occludin and junctional adhesion molecule-A (JAM-A) were detected by immunofluorescence. Sertoli cells isolated from Sprague-Dawley rats of postnatal day 10 were cultured in vitro and treated with TNFα. Cell proliferation rate was reflected by Cell Counting Kit-8 (CCK-8) and 5-ethynyl-2'-deoxyuridine (EdU) assay. Immunoblot and quantitative polymerase chain reaction were used to detect the expression of proliferating cell nuclear antigen, Fbxo4, and cyclin D1. Immunoprecipitation was used to detect the ubiquitination of cyclin D1 and the interaction between Fbxo4 and cyclin D1. Ammonium pyrrolidinedithiocarbamate (PDTC) was applied to detect the effect of nuclear factor kappaB (NFκB) activity inhibition on TNFα-induced Sertoli cell proliferation. The adenoviral recombinant plasmid containing rat Fbxo4 gene was constructed to investigate the effect of Fbxo4 overexpression on Sertoli cell proliferation promoted by TNFα. The in vivo experiment revealed a significant delay of blood-testis barrier maturation in pubertal rats caused by exogenous TNFα. TNFα (10ng/ml) treatment in vitro was found to promote the proliferation of immature Sertoli cells, accompanied with increased NFκB activity and cyclin D1 protein level. The level of Fbxo4 and ubiquitination of cyclin D1 were decreased after TNFα treatment. Inhibitor of NFκB or overexpression of Fbxo4 could both reverse the TNFα-induced proliferation of immature Sertoli cells, meanwhile restore the ubiquitin-proteasome system-dependent degradation of cyclin D1. Overexpression of Fbxo4 could not affect the activation of NFκB caused by TNFα. These results indicate that TNFα inhibits the ubiquitination and degradation of cyclin D1 through the NFκB pathway, thereby promoting the proliferation of immature Sertoli cell in vitro and inducing the delay of blood-testis barrier maturation in pubertal rats.

Endothelin A receptors contribute to senescence of brain microvascular endothelial cells.

Cellular senescence plays a pivotal role in the aging and progression of neurodegenerative diseases, including vascular cognitive impairment and dementia (VCID). In postmortem brains from individuals with VCID, endothelin-1 (ET-1) levels closely correlate with blood barrier breakdown and cerebral hypoperfusion. Brain microvascular endothelial cells (BMVECs), previously thought to have exclusively endothelin B receptors, also possess endothelin A (ETA) receptors; however, the functional significance of this receptor in BMVECs is not known. We hypothesize that ETA receptors mediate BMVEC senescence. Serum-starved human BMVECs (HBEC5i) were incubated with ET-1 (1µmol/L) in the presence/absence of ETA receptor antagonist BQ-123 (20µmol/L). Cells were collected for Western blot and quantitative real-time PCR analyses. Treatment of ET-1 increased protein expression of ETA receptor, while it was prevented by the ETA receptor antagonist. ET-1 increased p21, p16, p53, LIF1 and cyclin D1 protein levels, and β-galactosidase accumulation, which were prevented in the presence of ETA blockade. While there was no change in tight junction proteins, ET-1 decreased adherent junction protein vascular endothelial cadherin (VE-cadherin) levels. In conclusion, ET-1 upregulates ETA receptors in BMVECs in an autocrine manner and triggers the activation of senescence. These in vitro findings need to be further studied in vivo to establish the role of ETA receptors in the progression of endothelial senescence in VCID.

Photosynthetic modulation during the diurnal cycle in a unicellular diazotrophic cyanobacterium grown under nitrogen-replete and nitrogen-fixing conditions

Cyanobacteria are the only oxygenic photosynthetic organisms that can fix nitrogen. In diazotrophic cyanobacteria, the regulation of photosynthesis during the diurnal cycle is hypothesized to be linked with nitrogen fixation and involve the D1 protein isoform PsbA4. The amount of bioavailable nitrogen has a major impact on productivity in aqueous environments. In contrast to low- or nitrogen-fixing (−N) conditions, little data on photosynthetic regulation under nitrogen-replete (+ N) conditions are available. We compared the regulation of photosynthesis under −N and + N conditions during the diurnal cycle in wild type and a psbA4 deletion strain of the unicellular diazotrophic cyanobacterium Cyanothece sp. ATCC 51142. We observed common changes to light harvesting and photosynthetic electron transport during the dark in + N and −N conditions and found that these modifications occur in both diazotrophic and non-diazotrophic cyanobacteria. Nitrogen availability increased PSII titer when cells transitioned from dark to light and promoted growth. Under −N conditions, deletion of PsbA4 modified charge recombination in dark and regulation of PSII titer during dark to light transition. We conclude that darkness impacts the acceptor-side modifications to PSII and photosynthetic electron transport in cyanobacteria independently of the nitrogen-fixing status and the presence of PsbA4.

Crystal structures of photosystem II from a cyanobacterium expressing psbA2 in comparison to psbA3 reveal differences in the D1 subunit

Three psbA genes (psbA1, psbA2, and psbA3) encoding the D1 subunit of photosystem II (PSII) are present in the thermophilic cyanobacterium Thermosynechococcus elongatus and are expressed differently in response to changes in the growth environment. To clarify the functional differences of the D1 protein expressed from these psbA genes, PSII dimers from two strains, each expressing only one psbA gene (psbA2 or psbA3), were crystallized, and we analyzed their structures at resolutions comparable to previously studied PsbA1-PSII. Our results showed that the hydrogen bond between pheophytin/D1 (PheoD1) and D1-130 became stronger in PsbA2- and PsbA3-PSII due to change of Gln to Glu, which partially explains the increase in the redox potential of PheoD1 observed in PsbA3. In PsbA2, one hydrogen bond was lost in PheoD1 due to the change of D1-Y147F, which may explain the decrease in stability of PheoD1 in PsbA2. Two water molecules in the Cl-1 channel were lost in PsbA2 due to the change of D1-P173M, leading to the narrowing of the channel, which may explain the lower efficiency of the S-state transition beyond S2 in PsbA2-PSII. In PsbA3-PSII, a hydrogen bond between D1-Ser270 and a sulfoquinovosyl-diacylglycerol molecule near QB disappeared due to the change of D1-Ser270 in PsbA1 and PsbA2 to D1-Ala270. This may result in an easier exchange of bound QB with free plastoquinone, hence an enhancement of oxygen evolution in PsbA3-PSII due to its high QB exchange efficiency. These results provide a structural basis for further functional examination of the three PsbA variants.

PSAT176 Molecular Analysis of AMBRA1 as a Candidate Tumor Suppressor in Sporadic Parathyroid Adenomas

Abstract Primary hyperparathyroidism is a common endocrine disorder that is most often caused by a sporadic single-gland parathyroid adenoma. Currently, the only known and experimentally validated oncoprotein for parathyroid adenomas is cyclin D1. The overexpression of cyclin D1 protein has been reported in twenty to forty percent of cases. Eight percent of this overexpression can be explained by DNA rearrangement involving the cyclin D1 locus. The remaining molecular mechanisms behind this overexpression have yet to be identified. In this study, we explored a potential parathyroid tumorigenic mechanism that could increase cyclin D1 stability through a defect in the AMBRA1 E3 ligase adaptor, a molecule responsible for its degradation. Evidence that AMBRA1 may be a tumor suppressor gene includes the accumulation of cyclin D with hyperproliferation in AMBRA1-deficient cells, the context-dependent growth of tumors in Ambra1-deficient mice, and the finding that low levels of AMBRA1 correlated with poor clinical outcomes in The Cancer Genome Atlas (TCGA). We have therefore proceeded examine AMBRA1 for evidence of tumor suppressor-type inactivation within a cohort of parathyroid adenomas. Genomic DNA from twenty-six typical sporadic parathyroid adenomas was subjected to PCR-amplification of all eighteen coding exons of AMBRA1, followed by Sanger sequencing. Sequences were analyzed by comparison to the normal reference sequence, seeking to assess any observed intragenic or splice-site variants as potential or likely loss-of-function mutations. Thus far, with ninety-seven percent of the coding region fully sequenced, no intragenic AMBRA1 inactivating mutations were observed. Three known single nucleotide polymorphisms, considered non-pathogenic, were identified: missense variant c.3028G>T, and 3' UTR variants c.*491T>C and c.*576A>T, each in a single tumor. Future study of increased numbers of parathyroid adenomas is needed to exclude the possibility that AMBRA1 inactivation could play a driver role in a small percentage of cases, and the possibility that AMBRA1 might contribute less directly e.g. via secondarily altered expression also warrants investigation. Nonetheless, our current observations strongly suggest that AMBRA1 does not have a frequent role as a classic 2-hit tumor suppressor gene in sporadic parathyroid adenomas. Presentation: Saturday, June 11, 2022 1:00 p.m. - 3:00 p.m.

Exogenous trehalose protects photosystem II in heat-stressed wheat

Background: Photosystem II (PSII) is susceptible to heat stress. Plants naturally accumulate trehalose to improve stress tolerance. However, the mechanism by which trehalose affects PSII during heat stress is still unknown.
 Questions: How does trehalose affect PSII during heat stress?
 Studied species: Triticum aestivum L.
 Study site and dates: Shanghai, China. 2019-2021.
 Methods: Trehalose -pretreated wheat seedlings suffered from heat stress and their photosynthetic parameters were measured.
 Results: Heat stress caused a reduction in the photochemical efficiency of PSII, the electron transfer rate (ETR(II)), the quantum yield of regulated energy dissipationY(NPQ) and the coefficient of photochemical quenching (qP), but increased the quantum yield of non-regulated energy dissipation of PSII (Y[NO]). The shape of the fast chlorophyll fluorescence induction kinetics (OJIP) curve in the heat-stressed wheat was altered and the primary photochemistry maximum yield of PSII (Fv/Fo) and the PSII performance indicator PIabs were reduced. Accordingly, the activities of PSII and electron transport chain, the amount of ordered α-helix structures and the content of D1 protein also decreased. However, in trehalose-pretreated wheat, D1 protein and protein secondary structures of PSII were both protected, the electron transport activities of PSII and the whole chain were improved and greater fluorescence parameters values were maintained. Lower Y(NO) and more stable OJIP were obtained.
 Conclusions: Exogenous trehalose acted a vital role in the protection of the function of PSII, resulting in higherphotosynthetic capacity under heat stress.

Revealing the contribution of GbPR10.5D1 to resistance against Verticillium dahliae and its regulation for structural defense and immune signaling.

As an important family of pathogenesis-related (PR) proteins, the functional diversification and roles of PR10s in biotic stress have been well documented. However, the molecular basis of PR10s in plant defense responses against pathogens remains to be further understood. In the present study, we analyzed the phylogenetic relationship and function of a novel PR10 named GbPR10.5D1 in Sea-Island (or Pima or Egyptian) cotton (Gossypium barbadense L.), which has been identified as a Verticillium dahliae Kleb.-induced protein in a previous proteomics study. Phylogenetic analysis revealed that GbPR10.5D1, located on chromosome 2, is a unique member of GbPR10. The expression of GbPR10.5D1 was preferably in the root and induced upon V. dahliae infection. GbPR10.5D1 proteins were distributed in both nucleus and cytoplasm. GbPR10.5D1-virus-induced gene-silencing (VIGS) cotton plants were more susceptible to infection by V. dahliae, whereas overexpression (OE) of GbPR10.5D1 in cotton enhanced the resistance. By comparative transcriptome analysis between GbPR10.5D1-OE and wild-type (WT) plants and quantitative real-time polymerase chain reaction (qRT-PCR) verification, we found transcriptional activation of genes involved in cutin, suberine, and wax biosynthesis and mitogen-activated protein kinase (MAPK) signaling under normal conditions. Upon pathogen infection, defense signaling, fatty acid degradation, and glycerolipid metabolism were specifically activated in GbPR10.5D1-OE plants; biological processes (BPs), including glycolysis and gluconeogenesis, DNA replication, and cell wall organization, were specifically repressed in WT plants. Collectively, we proposed that GbPR10.5D1 possibly mediated lipid metabolism pathway to strengthen structural defense and activate defense signaling, which largely released the repression of cell growth caused by V. dahliae infection.

Effect of Mycotoxin Cytochalasin A on Photosystem II in Ageratina adenophora.

Biological herbicides have received much attention due to their abundant resources, low development cost, unique targets and environmental friendliness. This study reveals some interesting effects of mycotoxin cytochalasin A (CA) on photosystem II (PSII). Our results suggested that CA causes leaf lesions on Ageratina adenophora due to its multiple effects on PSII. At a half-inhibitory concentration of 58.5 μΜ (I50, 58.5 μΜ), the rate of O2 evolution of PSII was significantly inhibited by CA. This indicates that CA possesses excellent phytotoxicity and exhibits potential herbicidal activity. Based on the increase in the J-step of the chlorophyll fluorescence rise OJIP curve and the analysis of some JIP-test parameters, similar to the classical herbicide diuron, CA interrupted PSII electron transfer beyond QA at the acceptor side, leading to damage to the PSII antenna structure and inactivation of reaction centers. Molecular docking model of CA and D1 protein of A. adenophora further suggests that CA directly targets the QB site of D1 protein. The potential hydrogen bonds are formed between CA and residues D1-His215, D1-Ala263 and D1-Ser264, respectively. The binding of CA to residue D1-Ala263 is novel. Thus, CA is a new natural PSII inhibitor. These results clarify the mode of action of CA in photosynthesis, providing valuable information and potential implications for the design of novel bioherbicides.

4-Hydroxyphenylpyruvate dioxygenase (HPPD)-inhibiting herbicides: past, present, and future

AbstractThe herbicides that inhibit 4-hydroxyphenylpyruvate dioxygenase (HPPD) are primarily used for weed control in corn, barley, oat, rice, sorghum, sugarcane, and wheat production fields in the United States. The objectives of this review were to summarize 1) the history of HPPD-inhibitor herbicides and their use in the United States; 2) HPPD-inhibitor resistant weeds, their mechanism of resistance, and management; 3) interaction of HPPD-inhibitor herbicides with other herbicides; and 4) the future of HPPD-inhibitor-resistant crops. As of 2022, three broadleaf weeds (Palmer amaranth, waterhemp, and wild radish) have evolved resistance to the HPPD inhibitor. The predominance of metabolic resistance to HPPD inhibitor was found in aforementioned three weed species. Management of HPPD-inhibitor-resistant weeds can be accomplished using alternate herbicides such as glyphosate, glufosinate, 2,4-D, or dicamba; however, metabolic resistance poses a serious challenge, because the weeds may be cross-resistant to other herbicide sites of action, leading to limited herbicide options. An HPPD-inhibitor herbicide is commonly applied with a photosystem II (PS II) inhibitor to increase efficacy and weed control spectrum. The synergism with an HPPD inhibitor arises from depletion of plastoquinones, which allows increased binding of a PS II inhibitor to the D1 protein. New HPPD inhibitors from the azole carboxamides class are in development and expected to be available in the near future. HPPD-inhibitor-resistant crops have been developed through overexpression of a resistant bacterial HPPD enzyme in plants and the overexpression of transgenes for HPPD and a microbial gene that enhances the production of the HPPD substrate. Isoxaflutole-resistant soybean is commercially available, and it is expected that soybean resistant to other HPPD inhibitor herbicides such as mesotrione, stacked with resistance to other herbicides, will be available in the near future.

MiR-193a-5p Enhances the Radioresistance of Pancreatic Cancer Cells by Targeting ZFP57 and Activating the Wnt Pathway.

This study was to investigate whether miR-193a-5p and ZFP57 are involved in the radioresistance of pancreatic cancer and to explore its working mechanism. Pancreatic cancer tissues were harvested from patients who achieved CR (complete remission) and PR (partial remission) and those who achieved PD (progressive disease) and SD (stable disease). The mRNA and protein expressions of ZFP57 and miR-193a-5p were determined by RT-qPCR and WB (Western blot), respectively. For in vitro experiments, the parental BxPC-3 cell line was irradiated by X-ray at a total dose of 40 Gy to induce the irradiation-resistant subtype BxPC-3-RR. ZFP57 was downregulated in radioresistant pancreatic cancer cells. The results of dual-luciferase reporter gene assay, RNA pull-down assay, RT-qPCR, and WB confirmed that miR-193a-5p targeted ZFP57 and inhibited ZFP57 expression. The MTT assay and the colony formation assay showed that the radioresistant pancreatic cancer cells had higher viability and survival fraction. The results of WB indicated that in the radioresistant pancreatic cancer cells, the cyclin D1, Bax, CDk4, cleaved caspase-3, Bcl-2, and γ-H2AX proteins were upregulated to varying degrees. The results of the in vitro nude mouse experiment were consistent with those of in vivo experiments. According to the cell transfection and salvage experiments, miR-193a-5p down regulated ZFP57 after radiotherapy. As a result, the Wnt pathway was activated, which further induced radioresistance of pancreatic cancer cells. Our experiments showed that the miR-193a-5p/ZFP57/Wnt pathway mediated the radioresistance of pancreatic cancer cells, providing novel clues for the treatment of pancreatic cancer.

Magnetic Nanodroplets for Enhanced Deep Penetration of Solid Tumors and Simultaneous Magnetothermal-Sensitized Immunotherapy against Tumor Proliferation and Metastasis.

The central cells of solid tumors are more proliferative and metastatic than the marginal cells. Therefore, more intelligent strategies for targeting cells with deep spatial distributions in solid tumors remain to be explored. In this work, a biocompatible nanotheranostic agent with a lipid membrane-coated, Fe3 O4 and perfluoropentane (PFP)-loaded, cRGD peptide (specifically targeting the integrin αvβ3 receptor)-grafted, magnetic nanodroplets (MNDs) is developed. The MNDs exhibit excellent magnetothermal conversion and controllable magnetic hyperthermia (MHT) through alternating magnetic field regulation. Furthermore, MHT-mediated magnetic droplet vaporization (MDV) induces the expansion of the MNDs to transform them into ultrasonic microbubbles, increasing the permeability of tissue and the cell membrane via the ultrasound-targeted microbubble destruction (UTMD) technique and thereby promoting the deep penetration of MNDs in solid tumors. More importantly, MHT not only causes apoptotic damage by downregulating the expression of the HSP70, cyclin D1, and Bcl-2 proteins in tumor cells but also improves the response rate to T-cell-related immunotherapy by upregulating PD-L1 expression in tumor cells, thus inhibiting the growth of both primary and metastatic tumors. Overall, this work introduces a distinct application of nanoultrasonic biomedicine in cancer therapy and provides an attractive immunotherapy strategy for preventing the proliferation and metastasis of deeply distributed cells in solid tumors.

Deacylation of galactolipids decomposes photosystem II dimers to enhance degradation of damaged D1 protein.

Photosystem II (PSII) contains many lipid molecules that are essential for the function and maintenance of PSII. Under strong light conditions, PSII complexes are dynamically modified during the repair process; however, the molecular mechanism of the dynamic changes in the PSII structure is still unclear. In the present study, we investigated the role of a lipase in the repair of PSII in Synechocystis sp. PCC 6803. We identified a protein encoded by the sll1969 gene, previously named lipase A (lipA), in the Synechocystis sp. PCC 6803 genome as a candidate for the lipase involved in PSII repair. Recombinant protein expressed in Escherichia coli cells hydrolyzed fatty acids at the sn-1 position of monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol as well as triacylglycerol esterified with stearic acids. PSII repair in a disrupted mutant of the lipA gene was suppressed by the slow degradation of damaged D1 protein under strong light. The level of the PSII dimer remained higher in lipA mutant cells than wild-type (WT) cells under strong light. LipA protein was associated with the PSII dimer in vivo, and recombinant LipA protein decomposed PSII dimers purified from WT cells to monomers by reducing MGDG content in the PSII complex. These results indicate that LipA reacts with PSII dimers, dissociates them into monomers by digesting MGDG, and enhances D1 degradation during PSII repair.

Based on molecular docking to evaluate the protective effect of saponins from ginseng berry on D-gal-induced brain injury via multiple molecular mechanisms in mice

• To explore the effect of ginseng berries saponins on D-gal-induced brain injury. • The content of various saponins in ginseng berries was quantitatively analyzed. • Ginseng berry saponins can improve oxidative stress, apoptosis and cell aging. • Ginseng berry saponins may activate the SIRT1/Akt signaling pathway. The present study investigated the protective effects of crude saponins (ginsenosides) in berries in an brain injury model by intraperitoneal injection of D-gal (D-galactose). Our study showed that ginseng berry saponin (GBS) treatment significantly improved memory impairment in mice. Meanwhile, long-term exposure to D-gal reduces the expression of cholinergic modulators such as choline acetyltransferase (ChAT) and acetylcholinesterase (AChE), which can be significantly improved after administration. Furthermore, we found that GBS improved redox levels in the brain. In addition, TUNEL staining results showed that GBS inhibited D-Gal-induced apoptosis. Finally, we found that GBS treatment delayed cell aging by modulating the expression levels of cyclin D1 and senescence proteins p16 and p21, and further attenuated oxidative stress by activating SIRT1/Akt pathway. These results suggest that GBS can effectively treat D-Gal-induced brain damage and may be a potential therapeutic agent for slowing down the aging process.

Nanoplastic impacts on the foliar uptake, metabolism and phytotoxicity of phthalate esters in corn (Zea mays L.) plants.

Nanoplastic pollution in terrestrial plants is of increasing concern for its negative effects on living organisms. However, the impacts of nanoplastics on chemical processes and plant physiology of phthalate esters (PAEs) remain unclear. The present work offers insight into the foliar uptake, metabolism and phytotoxicity of two typical PAEs, namely, di-n-butyl phthalate (DBP) and di-(2-ethylhexyl) phthalate (DEHP), in corn (Zea mays L.) seedlings and the effects of amino-functionalized polystyrene nanoplastics (PSNPs-NH2). The presence of PSNPs-NH2 increased DBP and DEHP accumulation in the leaves by 1.36 and 1.32 times, respectively. PSNPs-NH2 also promoted the leaf-to-root translocation of DBP and DEHP, with the translocation factor increasing by approximately 1.05- and 1.16-fold, respectively. Furthermore, the addition of PSNPs-NH2 significantly enhanced the transformation of PAEs to their primary metabolites, mono-butyl phthalate and mono(2-ethylhexyl) phthalate in corn leaves and roots. The co-presence of PSNPs-NH2 and PAEs showed stronger impairment of photosystem II efficiency via the downregulation of transporter D1 protein, thus exhibiting a greater inhibitory effect on plant growth. Our findings reveal that nanoplastics promote the foliar uptake and transformation of PAE chemicals in crops and exacerbate their toxicity to crop plants, thereby threatening agricultural safety and human health.

Expression of Cell-Cycle Regulatory Proteins pRb, Cyclin D1, and p53 Is Not Associated with Recurrence Rates of Equine Sarcoids.

Simple SummaryExpression of cell-cycle regulatory proteins pRB, Cyclin D1, and p53 is not associated with recurrence rates of equine sarcoids. Sarcoids are common tumors of equids, and since the pathogenesis and Bovine Papillomavirus (BPV) mechanism of tumorigenesis is not fully understood yet, we investigated the expression of cell cycle regulatory proteins pRB, Cyclin D1, and p53 to understand whether BPV and Human Papillomavirus (HPV) share similar mechanisms of action. The expression pattern of these proteins in a series of BPV-positive equine sarcoids is different from HPV-induced tumors. Moreover, their expression is not correlated with the recurrence or occurrence of sarcoids in new body sites.Sarcoids are among the most common tumors diagnosed in equids; their association with bovine papillomaviruses (BPV) infection has been widely reported, but the mechanism of carcinogenesis has not been fully elucidated. To verify whether BPV infection causes dysregulation of the pRb-Cyclin D1-p16CDKN2A-p53 pathway as reported for human papillomavirus (HPV), the study employed immunohistochemistry to test 55 equine sarcoid biopsies for the expression of pRb, Cyclin D1, and p53 cell cycle regulatory proteins and to evaluate the proliferative rate through Ki67. High Cyclin D1 and pRb expression were observed in 51% and 80% of cases, respectively, while low expression was observed in 49% and 20% of cases, respectively. Significantly higher Ki67 proliferation indexes were observed in fibroblastic, nodular, and mixed sarcoids compared to the occult and verrucous. High proliferation was significantly associated with high Cyclin D1 expression. In contrast with previous studies, p53 positivity was not observed in the cases examined in this study. Moreover, follow-up analysis revealed that fibroblastic, mixed sarcoids were associated with significantly higher local recurrence rates while the verrucous subtype was associated with higher rates of new sarcoid development at distant sites.

1,25-dihydroxyvitamin D3 ameliorates high glucose-mediated proliferation, migration, and MCP-1 secretion of vascular smooth muscle cells by inhibiting MAPK phosphorylation

ObjectivesTo explore the impacts of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) on the proliferation, migration, and monocyte chemoattractant protein-1 (MCP-1) secretion of vascular smooth muscle cells (VSMCs) in a high glucose environment and its possible mechanism.MethodsWe extracted VSMCs from the thoracic aorta of a male Sprague–Dawley rats before culturing them in a 25-mM glucose-containing medium in the presence or absence of 1,25(OH)2D3 (10−9 –10−7 M). Cell proliferation was determined by bromodeoxyuridine incorporation assays. Subsequently, cell migratory capacity was examined by performing a transwell assay. An enzyme-linked immunosorbent assay was conducted to assess MCP-1 levels. Protein levels of matrix metalloproteinase-9 (MMP-9), mitogen-activated protein kinases (MAPKs), cyclin D1, and phosphorylated MAPKs were determined by immunoblotting.Results1,25(OH)2D3 significantly suppressed the proliferation, migration, and MCP-1 secretion of VSMCs mediated by high glucose in a dose-dependent manner, diminished the enhanced protein expression of MMP-9 and cyclin D1, and attenuated MAPK phosphorylation. The p38 inhibitor SB203580 and ERK1/2 inhibitor PD98059 suppressed high glucose-mediated upregulation of MMP-9 and cyclin D1 protein expression and MCP-1 secretion, respectively.Conclusions1,25(OH)2D3 ameliorates high glucose-mediated proliferation, migration, and MCP-1 secretion of VSMCs by inhibiting MAPK phosphorylation, implying a potential therapeutic approach using 1,25(OH)2D3 for diabetic macrovascular complications.

Theabrownin Alleviates Colorectal Tumorigenesis in Murine AOM/DSS Model via PI3K/Akt/mTOR Pathway Suppression and Gut Microbiota Modulation.

Colorectal cancer (CRC) is one of the most common and fatal cancers worldwide, yet therapeutic options for CRC often exhibit strong side effects which cause patients’ well-being to deteriorate. Theabrownin (TB), an antioxidant from Pu-erh tea, has previously been reported to have antitumor effects on non-small-cell lung cancer, osteosarcoma, hepatocellular carcinoma, gliomas, and melanoma. However, the potential antitumor effect of TB on CRC has not previously been investigated in vivo. The present study therefore aimed to investigate the antitumor effect of TB on CRC and the underlying mechanisms. Azoxymethane (AOM)/dextran sodium sulphate (DSS) was used to establish CRC tumorigenesis in a wild type mice model. TB was found to significantly reduce the total tumor count and improve crypt length and fibrosis of the colon when compared to the AOM/DSS group. Immunohistochemistry staining shows that the expression of the proliferation marker, Ki67 was reduced, while cleaved caspase 3 was increased in the TB group. Furthermore, TB significantly reduced phosphorylation of phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), and the downstream mechanistic target of rapamycin (mTOR)and cyclin D1 protein expression, which might contribute to cell proliferation suppression and apoptosis enhancement. The 16s rRNA sequencing revealed that TB significantly modulated the gut microbiota composition in AOM/DSS mice. TB increased the abundance of short chain fatty acid as well as SCFA-producing Prevotellaceae and Alloprevotella, and it decreased CRC-related Bacteroidceae and Bacteroides. Taken together, our results suggest that TB could inhibit tumor formation and potentially be a promising candidate for CRC treatment.

The Biomarker Like the Correlation between Vasculogenic Mimicry, Vascular Endothelial Cadherin, Sex-DeterminingRegion on Y-Box Transcription Factor 17, and Cyclin D1 in Oesophageal Squamous Cell Carcinoma.

Background This study aimed to explore the relationships between the sex-determining region on Y (SRY) box transcription factor 17 (SOX17), Cyclin D1, vascular endothelial cadherin (VE-cadherin), and vasculogenic mimicry (VM) in the occurrence and development of esophageal squamous cell carcinoma (ESCC). Methods The expressions of SOX17, Cyclin D1, and VE-cadherin, as well as VM, in tissues, were determined using immunohistochemistry. SOX17, Cyclin D1, and VE-cadherin mRNA in ESCC and their corresponding adjacent normal tissues were quantified using quantitative reverse transcription polymerase chain reaction analysis. Cell invasion, migration, and proliferation were determined after the silencing of VE-cadherin. SOX17, Cyclin D1, and VE-cadherin protein were quantified using Western blotting. Results The expression levels of SOX17, Cyclin D1, and VE-cadherin significantly correlated with the clinical characteristics of ESCC. After the VE-cadherin silencing, cell invasion, migration, and proliferation decreased, along with the Cyclin D1 levels, while the SOX17 levels increased. Conclusion SOX17, Cyclin D1, and VE-cadherin are involved in the development of ESCC.

​Exploring Inhibitory Potential of Curcumin against Multiple Targets Involved in the Cancer Progression, Metastasis and Apoptosis Pathways by in silico Molecular Docking

Background: Cancer is a multifactorial disease characterized by altered gene expression and one of the most leading causes of death in the world. Curcumin obtained from Curcuma longa known to exhibit anti-inflammatory, antioxidant as well as anti-cancer properties. The present study was undertaken to know different targets through which curcumin acts to elicit the antitumor effect. Methods: In this present study in silico interaction of curcumin with different cancer targets were analyzed by molecular docking using Biovia Discovery studio 4.0. Proteins were selected based on their role in cancer pathways which include transcription factor Nf-kB p50 and p65 subunit, Matrix metalloproteinase involved in invasion and metastasis, cell cycle regulator cyclin D and apoptotic factor Bax. Result: Curcumin found to interact with all the selected proteins with high dock score of 125.428, 94.778, 114.5,111.731,98.6844 with Nf-kB p50, p65 subunits, MMP9, Cyclin D1 and Bax proteins, respectively. Amino acid residues to which curcumin interact, number of hydrogen bonds and hydrogen bond length also predicted. RMSD (Root Mean Square Deviation) value was found to be one in all the interactions. Based on the molecular interaction studies and ADME (Absorption Distribution Metabolism Excretion) predictions it was found that curcumin can act as potent inhibitor of many cancer targets and can be used as a starting molecule for the design of anticancerous drug.

Expression of SASP, DNA Damage Response, and Cell Proliferation Factors in Early Gastric Neoplastic Lesions: Correlations and Clinical Significance.

The cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING)-mediated senescence-associated secretory phenotype (SASP) pathway has recently been identified in the suppression and promotion of cancers. However, its practical role in carcinogenesis remains to be comprehensively elucidated. Here, we describe an investigation analysing SASP activity and its correlations with DNA damage response (DDR), genomic mutations, and cell proliferation in gastric carcinogenesis among 30 cases with available endoscopic submucosal dissection (ESD) specimens of early neoplastic lesions (including low-grade dysplasia [LGD], high-grade dysplasia [HGD], and intramucosal carcinoma). The positive cells of senescence-associated β-galactosidase staining and cGAS, STING, interferon-regulatory factor 3 (IRF3), and signal transducer and activator of transcription 6 (STAT6) expression levels using immunostaining were elevated in HGD and in cancers. Similarly, increased expression of the Fanconi anemia group D2 (FANCD2) protein, tumour suppressor p53 binding protein 1 (TP53BP1), and replication protein A (RPA2) (i.e., primary DDR factors) was detected in HGD and in cancers; these increased expression levels were closely correlated with high expression of Ki67 and minichromosome maintenance complex component 7 (MCM7) proteins. Moreover, genomic mutations in TP53 gene were detected in 56.67% of the evaluated cases (17/30) using next-generation sequencing, and positive staining was verified in HGD and in cancers. Statistical analysis revealed that cell proliferation closely correlated with the expression of DDR factors, of which TP53BP1 was positively associated with SASP factors and IRF3 was positively correlated with cell proliferation. In addition, an analysis evaluating clinical features demonstrated that STAT6-positive cases showed a longer progression-free survival time than STAT6-negative cases. Our evaluation, conducted using a limited number of specimens, suggests SASP may be prevalent in early gastric neoplastic lesions and could be activated by accelerated cell proliferation-induced DDR. The clinical significance of SASP still needs to be determined.