Histology and histopathology Vol.39,nº10 (2024)
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- PublicationOpen AccessRNA-binding protein DND1 participates in migration, invasion, and EMT of prostate cancer cells by degrading CLIC4(Universidad de Murcia. Departamento de Biología Celular e Histología, 2024) Zhang, Wei; Xu, Qian; Shi, Chunmei; Chen, Xinfeng; Shen, Cheng; Zhang, Yong; Zheng, Bing; Zhu, HuaDead-End 1 (DND1) is an RNA-binding protein (RBP) with regulatory functions in multiple cancers, including gastric and colorectal. Nevertheless, the role that DND1 plays in prostatic cancer (PCa) as well as the hidden molecular mechanism is still obscure. The gene expression of DND1 and survival analyses in PCa were analyzed by the UALCAN database. Expression of DND1 and chloride intracellular channel 4 (CLIC4) were detected by qRT-PCR and western blot analysis. The Cell Counting Kit-8 assay and EDU staining were employed for the estimation of cell viability. The capabilities of cells to migrate and invade were appraised by the wound healing assay as well as the Transwell assay, while epithelial-mesenchymal transition (EMT) was measured by immunofluorescence and western blot assay. The interaction of DND1 and CLIC4 was predicted by PCTA, linkedomics, and RPISeq databases. It was discovered that DND1 expression was elevated in PCa cells. DND1 silencing had suppressive impacts on the proliferative, migrative, and invasive capabilities as well as EMT in DU145 and 22Rv1 cells. Mechanistically, bioinformatic analysis demonstrated that DND1 was negatively correlated with CLIC4 and that DND1 protein could bind to CLIC4 mRNA. Additionally, the CLIC4 level was reduced in PCa cells. CLIC4 depletion countervailed the suppressive impacts of DND1 deficiency on the capabilities of DU145 and 22Rv1 cells to proliferate, migrate, and invade as well as the process of EMT. These results suggested that DND1 silencing repressed the proliferation, migration, invasion, and EMT in PCa by regulating the mRNA level of CLIC4.
- PublicationOpen AccessThe immune microenvironment of cancer of the uterine cervix(Universidad de Murcia. Departamento de Biología Celular e Histología, 2024) Mastrogeorgiou, Michail; Chatzikalil, Elena; Theocharis, Stamatios; Papoudou-Bai, Alexandra; Péoc’h, Michel; Mobarki, Mousa; Karpathiou, GeorgiaWhile several treatment choices exist for cervical cancer, such as surgical therapy, chemotherapy, and radiotherapy, some patients will still show poor prognosis. HPV infection is a principal factor for cervical cancer development, from early inflammation to proliferation, angiogenesis, and neoplastic growth. While HPV T-cell responses exist, the tumor seems to evade the immune system upon its tolerance. The latter suggests the existence of a confluent tumor microenvironment responsible for the evasion tactics employed by the neoplasm. Therefore, novel biomarkers governing prognosis and treatment planning must be developed, with several studies tackling the significance of the tumor microenvironment in the genesis, development, proliferation, and overall response of cervical cancer during neoplastic processes. This review aims to analyze and contemplate the characteristics of the tumor microenvironment and its role in prognosis, progression, evasion, and invasion, including therapeutic outcome and overall survival.
- PublicationOpen AccessFemale fertility and the mammalian egg’s zona pellucida(Universidad de Murcia. Departamento de Biología Celular e Histología, 2024) Wassarman, Paul M.; Litscher, Eveline S.All mammalian eggs are surrounded by a relatively thick extracellular matrix (ECM) or zona pellucida (ZP) to which free-swimming sperm bind in a species-restricted manner during fertilization. The ZP consists of either three (e.g., Mus musculus) or four (e.g., Homo sapiens) glycosylated proteins, called ZP1-4. These proteins are unlike those found in somatic cell ECM, are encoded by single-copy genes on different chromosomes, and are well conserved among different mammals. Mammalian ZP proteins are synthesized as polypeptide precursors by growing oocytes that will become ovulated, unfertilized eggs. These precursors are processed to remove a signal-sequence and carboxyterminal propeptide and are secreted into the extracellular space. Secreted ZP proteins assemble into long, crosslinked fibrils that exhibit a structural repeat due to the presence of ZP2-ZP3 dimers every 140 Å or so along fibrils. Fibrils are crosslinked by ZP1 and are oriented either perpendicular, parallel, or randomly to the plasma membrane of eggs depending on their position in the ZP. Free-swimming mouse sperm recognize and bind to ZP2 or ZP3 that serve as sperm receptors. Acrosome-intact sperm bind to ZP3 oligosaccharides and acrosome-reacted sperm bind to ZP2 polypeptide. ZP fibrils fail to assemble in the absence of either nascent ZP2 or ZP3 and results in mouse eggs that lack a ZP and female infertility. Gene sequence variations in genes encoding ZP1-4 result in human eggs that lack a ZP or have an abnormal ZP and female infertility. These and other features of the mouse and human egg’s ZP are discussed here.
- PublicationOpen AccessThe potential of EZH2 expression to facilitate treatment choice in stage II colorectal adenocarcinoma(Universidad de Murcia. Departamento de Biología Celular e Histología, 2024) Zhu, Xiaoqun; He, Lu; Zheng, Zhong; Wang, Ya; Yang, Jun; Zhang, Biao; Wang, Chaoshan; Li, ZhiwenBackground. The current selection criteria of patients with stage II colorectal carcinoma (CRC) suitable for adjuvant therapy are not satisfactory. Enhancer of zeste homolog 2 (EZH2) has been demonstrated to be over-expressed in CRC. However, data regarding the role of EZH2 in CRC survival remains controversial, and little is known about it in stage II CRC. Thus, we conducted this study to investigate the clinical significance of EZH2 expression in stage II CRC. Methods. Cases with stage II CRC resected between 2015 and 2018 were retrospectively reviewed. EZH2 expression was analyzed by immunohistochemistry using tissue microarrays. The relationship between EZH2 expression and clinicopathological variables was analyzed. Survival curves were estimated by the KaplanMeier approach. Results. We found high EZH2 expression in 134 of 221 analyzable stage II tumors (60.63%). No significant associations were observed between EZH2 expression and common clinicopathological factors. Survival analyses showed that cases receiving surgery alone had inferior overall survival (OS) than those receiving surgery and chemotherapy (P=0.0075) in stage II CRC with high EZH2 expression, however, metastasis-free survival (MFS) was similar between these two subgroups. Treatment choice had no impact on the survival of stage II CRC with low EZH2 expression. Conclusion. The OS of stage II CRC with high EZH2 expression improved more strikingly with surgery and adjuvant chemotherapy than with surgery alone, which suggests the potential of EZH2 expression as a biomarker to help identify a subgroup of early-stage CRC benefiting from surgery and adjuvant chemotherapy. More large-scale studies are warranted to corroborate this finding and to further evaluate the predictive nature of EZH2.
- PublicationOpen AccessBiological action of bleaching agents on tooth structure: A review(Universidad de Murcia. Departamento de Biología Celular e Histología, 2024) Bragança Aragão, Walessa Alana; Santos Chemelo, Victória; de Melo Alencar, Cristiane; Martins Silva, Cecy; Pessanha, Sofia; Reis, Alessandra; de Souza Rodrigues, Renata Duarte; Rodrigues Lima, RafaelThe use of bleaching agents to remove stains is one of the main dental procedures to improve the aesthetics of teeth. This review presents the main agents used for tooth whitening, existing clinical protocols, and the structural changes that may occur through their use. The main bleaching agents consist of hydrogen peroxide and carbamide peroxide, which are used in bleaching techniques for vital teeth. These techniques can be performed in the office by a professional or by the individual in a home environment under professional guidance. Bleaching agents come in a variety of concentrations and there are over-the-counter products available on the market with lower concentrations of hydrogen peroxide. Due to the chemical characteristics of the agents, changes in the organic and inorganic content of the tooth structure can be observed. These changes are related to morphological changes characterized by increased permeability and surface roughness, such changes compromise the mechanical resistance of the tooth. Furthermore, bleaching agents can promote molecular changes after reaching the dental pulp, resulting in oxidative stress of pulp cells and the release of proinflammatory mediators. Despite the bleaching effectiveness, tooth sensitivity is considered the main side effect of use. Therefore, among the heterogeneity of protocols, those that used the bleaching agent for a prolonged time and in lower concentrations presented more harmful effects on the tooth structure.
- PublicationOpen AccessHistological and morphometrical evaluation of the urethral wall after bioresorbable stent implantation in male New Zealand White Rabbits: A preliminary study(Universidad de Murcia. Departamento de Biología Celular e Histología, 2024) Skonieczna-Kurpiel, Joanna; Madej, Jan P.; Klekiel, Tomasz; Mackiewicz, Agnieszka; Będziński, Romuald; Noszczyk-Nowak, Agnieszka; Piasecki, TomaszThe aim of the study was the histological and morphometrical evaluation of the urethral wall at three time points after bioresorbable stent implantation in male New Zealand White Rabbits. The research was performed on 26 male New Zealand White rabbits aged 3-4 months and weighing 2.1-3.0 kg. Two models of bioresorbable sodium alginate-based stents were developed and implanted into the urethral lumen for one (T1), three (T3), and six weeks (T6). Sections of 5 µm thickness were cut from the urethra at intervals of 2 mm. The sliced sections were stained with hematoxylin-eosin (H&E), Van Gieson's (VG), Von Kossa, and Movat– Russell modified pentachrome (MOVAT) staining methods. The study provided valuable information for future models of urethral stents. The first model of the stent failed to fit the requirements due to inadequate mechanical properties. It curled up on itself losing the ability to adhere to the animals’ urethra and was bioresorbed three weeks after implantation. The more rigid no. 2 stent was effective in widening the urethral lumen but did not biodegrade during the experiment. A comprehensive assessment of the second model’s properties of biosorption and biointegration requires an extended observation of at least 12 months for an in depth morphological analysis. Stent migration is not likely to be caused solely by the mechanical properties of the urethra or urinary flow but mainly by muscle contraction of the organ wall.
- PublicationOpen AccessColorectal cancer histopathology image analysis: A comparative study of prognostic values of automatically extracted morphometric nuclear features in multispectral and red-blue-green imagery(Universidad de Murcia. Departamento de Biología Celular e Histología, 2024) Liu, Wenlou; Qu, Aiping; Yuan, Jingping; Wang, Linwei; Chen, Jiamei; Zhang, Xiuli; Wang, Hongmei; Han, Zhengxiang; Li, Yan
- PublicationOpen AccessUpregulation of LY6K induced by FTO-mediated demethylation promotes the tumorigenesis and metastasis of oral squamous cell carcinoma via CAV-1-mediated ERK1/2 signaling activation(Universidad de Murcia. Departamento de Biología Celular e Histología, 2024) Xu, Chen; Gong, Rujuan; Yang, HaibingLymphocyte antigen 6 complex locus K (LY6K) has been demonstrated to play a significant role in cancers and identified as a therapeutic biomarker for head and neck squamous cell carcinoma. However, the role of LY6K in oral squamous cell carcinoma (OSCC) has not been explored. The current study discovered that LY6K was aberrantly upregulated in OSCC cell lines and tissues and that high LY6K expression significantly correlated with poorer survival of OSCC patients. Through stable knockdown of LY6K, we found that the growth, colony formation, migration, and invasion of OSCC cells were substantially suppressed. In addition, tumor growth and lung metastasis in vivo were effectively inhibited by LY6K depletion. Mechanically, LY6K binds with CAV-1 and activates CAV-1-mediated MAPK/ERK signaling to exert its oncogenic effects on OSCC. In addition, LY6K expression in OSCC was discovered to be regulated by FTO-mediated RNA N6- methyladenosine (m6A) modification in an IGF2BP1- dependent manner. Generally, LY6K expression was upregulated by FTO-mediated demethylation in OSCC, which promoted the tumorigenesis and metastasis of OSCC via activating the CAV-1-mediated ERK1/2 signaling pathway
- PublicationOpen AccessNeoastilbin ameliorates sepsis-induced liver and kidney injury by blocking the TLR4/NF-κB pathway(Universidad de Murcia. Departamento de Biología Celular e Histología, 2024) Xu, Ruiming; Wang, Dawei; Shao, Zhengyi; Li, Xiangbo; Cao, QiumeiSepsis frequently causes systemic inflammatory response syndrome and multiple organ failure in patients. Neoastilbin (NAS) is a flavonoid that plays vital functions in inflammation. This work aims to investigate the protective effects of NAS against sepsisinduced liver and kidney injury and elucidate its underlying mechanisms. The mouse model was established using cecal ligation puncture (CLP) induction. NAS was given to mice by gavage for 7 consecutive days before surgery. Liver and kidney function, oxidative stress, and inflammatory factors in serum or tissues were examined by ELISA or related kits. The expression of relevant proteins was assessed by Western blot. Hematoxylin and eosin and/or periodic acid-Schiff staining revealed that NAS ameliorated the pathological damage in liver and kidney tissues of CLPinduced mice. NAS improved liver and kidney functions, as evidenced by elevated levels of blood urea nitrogen, Creatinine, ALT, and AST in the serum of septic mice. TUNEL assay and the expression of Bcl-2 and Bax showed that NAS dramatically reduced apoptosis in liver and renal tissues. NAS treatment lowered the levels of myeloperoxidase and malondialdehyde, while elevated the superoxide dismutase content in liver and kidney tissues of CLPinduced mice. The levels of inflammatory cytokines (IL6, TNF-α, and IL-1β) in the serum and both tissues of CLP-injured mice were markedly decreased by NAS. Mechanically, NAS downregulated TLR4 expression and inhibited NF-κB activation, and overexpression of TLR4 reversed the protective effects of NAS against liver and kidney injury. Collectively, NAS attenuated CLP-induced apoptosis, oxidative stress, inflammation, and dysfunction in the liver and kidney by restraining the TLR4/NF-κB pathway.
- PublicationOpen AccessFunctional mechanism of baicalein in alleviating severe acute pancreatitis-acute lung injury by blocking the TLR4/MyD88/TRIF signaling pathway(Universidad de Murcia. Departamento de Biología Celular e Histología, 2024) Yang, Qingjing; Yue, Chao; Huang, Xing; Wang, Zihe; Li, Zhenlu; Hu, Weiming; Lu, Huiminy. Severe acute pancreatitis-acute lung injury (SAP-ALI) is a disease with high mortality. This study aims to explore the mechanism of baicalein on SAP-ALI in rats by blocking toll-like receptor-4 (TLR4)/myeloid differentiation primary response gene 88 (MyD88)/TIRdomain-containing adapter-inducing interferon-β (TRIF) signal pathway. The SAP-ALI rat model was established by intraperitoneal injection of 3% pentobarbital sodium (30 mg/kg), with pancreas and intestines turned over, injected with 3.5% sodium taurocholate backward into the bile-pancreatic duct at 0.1 mL/100 g for 12h, and treated with baicalein, lipopolysaccharide (LPS), miR182 agomir, or miR-182 antagomir. The TLR4/MyD88/ TRIF pathway was activated using LPS in SAP-ALI rats after baicalein treatment. Baicalein attenuated inflammatory cell infiltration, alveolar wall edema, decreased W/D ratio and levels of TLR4, MyD88, and TRIF in the lung tissues, reduced levels of inflammatory factors in pancreatic and lung tissues and BALF, diminished ROS, and elevated GSH, SOD and CAT in pancreatic and lung tissues of SAP-ALI rats. Activation of the TLR4/MyD88/TRIF pathway partly abrogated baicalein-mediated improvements in inflammation and oxidative stress in SAP-ALI rats. miR-182 targeted TLR4. miR-182 suppressed inflammation and oxidative stress in SAP-ALI rats by targeting TLR4. Inhibition of miR-182 partly nullified baicalein-mediated attenuation on inflammation and oxidative stress in SAP-ALI rats. In conclusion, baicalein can inhibit the TLR4/MyD88/ TRIF pathway and alleviate inflammatory response and oxidative stress in SAP-ALI rats by upregulating miR182 and suppressing TLR4, thus ameliorating SAP-ALI.
- PublicationOpen AccessHSP47 expression in the hamster Sertoli cell: An immunohistochemical study(Universidad de Murcia. Departamento de Biología Celular e Histología, 2024) Seco-Rovira, Vicente; Serrano-Sánchez, María Isabel; Beltrán-Frutos, Ester; Martínez-Hernández, Jesús; Ferrer, Concepción; Pastor, Luis MiguelHSP47, a chaperone whose main function is the maturation of collagen molecules, is considered a marker of fibrotic diseases. Increased collagen synthesis in the testis has been associated with various pathologies leading to seminiferous tubule regression. Our aim was to study whether HSP47 is expressed in hamster Sertoli cells both in the adult and in two physiological situations of seminiferous tubule atrophy: irreversible testicular ageing and testicular regression due to short photoperiod (reversible). Eighteen animals were divided as follows: a group of 6 young animals aged 6 months, a group of 6 animals aged 24 months, which were exposed to a long photoperiod, and a final group of 6 young animals subjected to a short photoperiod. Testicular samples were fixed in methacarn and an immuno-histochemical technique was used to detect HSP47. A semiquantitative study of this protein expression was performed between tubular sections of aged animals with complete spermatogenesis and arrested spermatogenesis and tubular sections with arrest spermatogenesis of photoinhibited testes. Sertoli cells were positive for HSP47, the intensity being greater in tubular sections with arrested spermatogenesis in both aged and photoinhibited animals. Semiquantitative analysis corroborated this observation in the sense that the expression of this protein differed according to the functional state of the seminiferous tubules. Thus, the ratio of immunoreactivity was significantly higher in tubular sections with arrested spermatogenesis in aged animals compared with regressed animals, and in the latter compared with those whose tubular sections showed complete spermatogenesis. In conclusion, HSP47 expression in Sertoli cells was found for the first time in mammals. Moreover, increased expression seemed to be related to the degree of seminiferous atrophy epithelium and to the reversible or nonreversible physiological state of this epithelium.