Apoptosis and Desquamation of Urothelial Cells in Tissue Remodeling During Rat Postnatal Development

Postnatal rat urothelium was studied from day 0 to day 14, when intense cell loss as part of tissue remodeling was expected. The morphological and biochemical characteristics of urothelial cells in the tissue and released cells were investigated by light and electron microscopy, by terminal deoxynucleotidyl transferase–mediated dUTP nick end labeling (TUNEL) assay, by annexin V/propidium iodide assay, and by immunofluorescent detection of active caspases and tight-junction protein occludin. Intense apoptosis and massive desquamation were detected between postnatal days 7 and 10. During this period, active caspases and TUNEL-positive cells were found in the urothelium. Disassembled cell–cell junctions were detected between cells. The majority of desquamated cells expressed no apoptotic cell morphology, but were active caspase positive and TUNEL positive. Ann+/PI- apoptotic bodies and desquamated Ann+/PI+ cells were detected in the lumen. These results indicate that apoptosis and desquamation participate in urothelial cell loss in the rat early postnatal period, indispensable for fast urothelial remodeling during development.

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114 PORCINE EMBRYO FRAGMENTATION, DEVELOPMENT AND APOPTOSIS: A CONFOCAL MICROSCOPY STUDY

Reproduction Fertility and Development ◽

10.1071/rdv17n2ab114 ◽

2005 ◽

Vol 17 (2) ◽

pp. 207

Author(s):

B. Mateusen ◽

A. Van Soom ◽

D. Maes ◽

A. de Kruif

Keyword(s):

Confocal Microscopy ◽

Apoptotic Cell ◽

Annexin V ◽

Terminal Deoxynucleotidyl Transferase ◽

Apoptotic Cells ◽

Blastocyst Development ◽

Cell Stage ◽

Developmental Arrest ◽

Morula Stage ◽

Embryonic Arrest

The relationship between embryonic fragmentation, embryonic arrest, and apoptosis has been the subject of some controversy (Hardy K 1999 Rev. Reprod. 4, 125–134). In order to investigate possible links, in vivo-produced, in vitro-cultured porcine embryos (n = 132) were scored for developmental stage and fragmentation at 7 days post insemination (dpi) and processed for propidium iodide and annexin V labelling. After fixation, embryos were processed for terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL). Using confocal microscopy, a cell was categorized apoptotic if (i) it had a fragmented or condensed nucleus, (ii) the cell membrane was annexin V-positive, and (iii) the nucleus was TUNEL labelled. An apoptotic cell ratio (ACR) was determined as the percentage of apoptotic cells per embryo. Differences in the % of fragmented and apoptotic embryos and correlations were analyzed using chi-square. Logistic regression was used to compare the average fragmentation % and the ACR. Sixty-one embryos (46%) arrested during the culture period, with 8 embryos arresting before or at the 4-cell stage. Significantly more arrested embryos were fragmented compared to embryos that were blastocysts at 7 dpi. Also, the average fragmentation percentage was significantly higher for arrested embryos compared to blastocysts. The correlation detected between developmental arrest and fragmentation was 0.60 (P < 0.05). None of the embryos without fragmentation had cells categorized as apoptotic, whereas 50 out of 55 embryos with fragmentation possessed apoptotic cells, which led to a correlation of 0.87 (P < 0.01) between fragmentation and apoptosis. The percentage of embryos with apoptotic cells was significantly higher for embryos arrested during the 5-cell to the morula stage compared to embryos that arrested before or at the 4-cell stage and embryos with blastocyst development at 7 dpi. The average ACR of embryos arrested during the 5-cell to the morula stage was significantly higher compared to the average ACR of blastocysts at 7 dpi. The correlation detected between the developmental arrest, during the 5-cell to the morula stage period and apoptosis was 0.57 (P < 0.01). Taken together, significant correlations between fragmentation, developmental arrest and apoptosis were detected. However, the association between embryonic arrest and apoptosis could be established only for embryos arrested after embryonic genome activation.

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CD74 in Apoptotic Macrophages Is Associated with Inflammation, Plaque Progression and Clinical Manifestations in Human Atherosclerotic Lesions

Metabolites ◽

10.3390/metabo12010054 ◽

2022 ◽

Vol 12 (1) ◽

pp. 54

Author(s):

Wei Li ◽

Nargis Sultana ◽

Linda Yuan ◽

Claes Forssell ◽

Xi-Ming Yuan

Keyword(s):

Clinical Symptoms ◽

Plaque Rupture ◽

Apoptotic Cell ◽

Clinical Manifestations ◽

Annexin V ◽

Statin Treatment ◽

Terminal Deoxynucleotidyl Transferase ◽

High Density Lipoproteins ◽

Atherosclerotic Lesions ◽

Thrombin Receptors

The aim of this study was to investigate whether CD74 levels in atherosclerotic lesions are associated with inflammation, apoptosis, plaque severity, and clinical symptoms among patients with carotid atherosclerosis. We further studied whether CD74 expression is associated with apoptosis in macrophages induced by 7ketocholesterol (7keto). Sixty-one carotid samples (39 males and 22 females) were immunostained with macrophages, smooth muscle cells, CD74, ferritin, TUNEL (Terminal deoxynucleotidyl transferase dUTP nick end labeling), and thrombin receptors. Double immunocytochemistry of CD74 and caspase 3 or CD74 and Annexin V was performed on THP-1 macrophages exposed to 7keto. In human carotid plaques, CD74 expression is lesion-dependently increased and is associated with necrotic core formation and plaque rupture, clinical symptoms, macrophage apoptosis, ferritin, and thrombin receptors. CD74 levels were inversely correlated to high-density lipoproteins and statin treatment, and positively correlated to triglycerides. In THP-1 macrophages, 7keto induced a significant increase in levels of CD74, ferritin, and apoptotic cell death. This study suggests that CD74 in apoptotic macrophages is linked to inflammation and thrombosis in progression of human atherosclerotic plaques, lipid metabolism, and clinical manifestation in atherosclerosis. Surface CD74 in apoptotic macrophages and ferritin production induced by oxidized lipids may contribute to inflammation and plaque vulnerability in atherosclerosis.

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Carfilzomib in Combination with Bortezomib Enhances Apoptotic Cell Death in B16-F1 Melanoma Cells

Biology ◽

10.3390/biology10020153 ◽

2021 ◽

Vol 10 (2) ◽

pp. 153

Author(s):

Min Seung Lee ◽

So Hyun Lim ◽

Ah-Ran Yu ◽

Chi Yeon Hwang ◽

Insug Kang ◽

...

Keyword(s):

Er Stress ◽

Dose Reduction ◽

Molecular Mechanisms ◽

Apoptotic Cell ◽

Proteasome Inhibitors ◽

Annexin V ◽

Fluorescein Isothiocyanate ◽

Melanoma Cells ◽

Eif2α Phosphorylation ◽

Pharmacological Interactions

Proteasome inhibitors, such as bortezomib (BZ) and carfilzomib (CFZ), have been suggested as treatments for various cancers. To utilize BZ and/or CFZ as effective therapeutics for treating melanoma, we studied their molecular mechanisms using B16-F1 melanoma cells. Flow cytometry of Annexin V-fluorescein isothiocyanate-labeled cells indicated apoptosis induction by treatment with BZ and CFZ. Apoptosis was evidenced by the activation of various caspases, including caspase 3, 8, 9, and 12. Treatment with BZ and CFZ induced endoplasmic reticulum (ER) stress, as indicated by an increase in eIF2α phosphorylation and the expression of ER stress-associated proteins, including GRP78, ATF6α, ATF4, XBP1, and CCAAT/enhancer-binding protein homologous protein. The effects of CFZ on ER stress and apoptosis were lower than that of BZ. Nevertheless, CFZ and BZ synergistically induced ER stress and apoptosis in B16-F1 cells. Furthermore, the combinational pharmacological interactions of BZ and CFZ against the growth of B16-F1 melanoma cells were assessed by calculating the combination index and dose-reduction index with the CompuSyn software. We found that the combination of CFZ and BZ at submaximal concentrations could obtain dose reduction by exerting synergistic inhibitory effects on cell growth. Moreover, this drug combination reduced tumor growth in C57BL/6 syngeneic mice. Taken together, these results suggest that CFZ in combination with BZ may be a beneficial and potential strategy for melanoma treatment.

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Indole-3-carbinol inhibits the proliferation of colorectal carcinoma LoVo cells through activation of the apoptotic signaling pathway

Human & Experimental Toxicology ◽

10.1177/09603271211021475 ◽

2021 ◽

pp. 096032712110214

Author(s):

JY Lee ◽

HM Lim ◽

CM Lee ◽

S-H Park ◽

MJ Nam

Keyword(s):

Colorectal Carcinoma ◽

Cancer Cells ◽

Inhibitory Activity ◽

Annexin V ◽

Fluorescein Isothiocyanate ◽

Cell Counting ◽

Terminal Deoxynucleotidyl Transferase ◽

Growth Inhibitory ◽

Growth Inhibitory Activity ◽

Lovo Cells

Indole-3-carbinol (I3C) is a phytochemical that exhibits growth-inhibitory activity against various cancer cells. However, there are limited studies on the effects of I3C on colon cancer cells. In this study, the growth-inhibitory activity of I3C against the human colorectal carcinoma cell line (LoVo) was examined. The results of the 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide, colony formation, and cell counting assays revealed that I3C suppressed the proliferation of LoVo cells. Microscopy and wound-healing analyses revealed that I3C affected the morphology and inhibited the migration of LoVo cells, respectively. I3C induced apoptosis and DNA fragmentation as evidenced by the results of fluorescein isothiocyanate-conjugated annexin V staining and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling assay, respectively. Additionally, I3C arrested the cell cycle at the G0/G1 phase and enhanced the reactive oxygen species levels. Western blotting analysis revealed that treatment with I3C resulted in the activation of apoptotic proteins, such as poly(ADP-ribose) polymerase, caspase-3, caspase-7, caspase-9, Bax, Bim, and p53 in LoVo cells. These results indicate that I3C induces apoptosis in LoVo cells by upregulating p53, leading to the activation of Bax and caspases. Taken together, I3C exerts cytotoxic effects on LoVo cells by activating apoptosis.

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Phytochemicals from Ajwa dates pulp extract induce apoptosis in human triple-negative breast cancer by inhibiting AKT/mTOR pathway and modulating Bcl-2 family proteins

Scientific Reports ◽

10.1038/s41598-021-89420-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Mohsin Ali Khan ◽

Sahabjada Siddiqui ◽

Imran Ahmad ◽

Romila Singh ◽

Durga Prasad Mishra ◽

...

Keyword(s):

Breast Cancer ◽

Triple Negative Breast Cancer ◽

Triple Negative ◽

Mononuclear Cells ◽

Apoptotic Cell ◽

Human Peripheral Blood ◽

Annexin V ◽

Phoenix Dactylifera ◽

Mtor Pathway ◽

Peripheral Blood Mononuclear

AbstractAjwa dates (Phoenix dactylifera L.) have been described in traditional and alternative medicine to provide several health benefits, but their mechanism of apoptosis induction against human triple-negative breast cancer MDA-MB-231 cells remains to be investigated. In this study, we analyzed the phytoconstituents in ethanolic Ajwa Dates Pulp Extract (ADPE) by liquid chromatography-mass spectrometry (LC–MS) and investigated anticancer effects against MDA-MB-231 cells. LC–MS analysis revealed that ADPE contained phytocomponents belonging to classes such as carbohydrates, phenolics, flavonoids and terpenoids. MTT assay demonstrated statistically significant dose- and time-dependent inhibition of MDA-MB-231 cells with IC50 values of 17.45 and 16.67 mg/mL at 24 and 48 h, respectively. Hoechst 33342 dye and DNA fragmentation data showed apoptotic cell death while AO/PI and Annexin V-FITC data revealed cells in late apoptosis at higher doses of ADPE. More importantly, ADPE prompted reactive oxygen species (ROS) induced alterations in mitochondrial membrane potential (MMP) in ADPE treated MDA-MB-231 cells. Cell cycle analysis demonstrated that ADPE induced cell arrest in S and G2/M checkpoints. ADPE upregulated the p53, Bax and cleaved caspase-3, thereby leading to the downregulation of Bcl-2 and AKT/mTOR pathway. ADPE did not show any significant toxicity on normal human peripheral blood mononuclear cells which suggests its safe application to biological systems under study. Thus, ADPE has the potential to be used as an adjunct to the mainline of treatment against breast cancer.

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Indirubin exerts anticancer effects on human glioma cells by inducing apoptosis and autophagy

AMB Express ◽

10.1186/s13568-020-01107-2 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Zhaohui Li ◽

Han Wang ◽

Jun Wei ◽

Liang Han ◽

Zhigang Guo

Keyword(s):

Apoptotic Cell ◽

Treatment Strategies ◽

Glioma Cells ◽

Annexin V ◽

Metastatic Potential ◽

Human Glioma ◽

Human Glioma Cells ◽

Efficient Treatment ◽

Cell Percentage ◽

Anticancer Effects

Abstract Glioma causes significant mortality across the world and the most aggressive type of brain cancer. The incidence of glioma is believed to increase in the next few decades and hence more efficient treatment strategies need to be developed for management of glioma. Herein, we examined the anticancer effects of Indirubin against a panel of human glioma cells and attempted to explore the underlying mechanisms. The results of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay showed that Indirubin could inhibit the growth of all the glioma cells but the lowest IC50 of 12.5 µM was observed against the U87 and U118 glioma cells. Additionally, the cytotoxic effects of Indirubin were comparatively negligible against the normal astrocytes with an IC50 of > 100 µM. Investigation of mechanism of action, revealed that Indirubin exerts growth inhibitory effects on the U87 and U118 glioma cells by autophagic and apoptotic cell death. Annexin V/PI staining assay showed that apoptotic cell percentage increased dose dependently. Apoptosis was associated with increase in Bax decrease in Bcl-2 expressions. Additionally, the expression of autophagic proteins such as LC3II, ATG12, ATG15 and Beclin 1 was also increased. Wound heal assay showed that Indirubin caused remarkable decrease in the migration of the U87 and U118 cells indicative of anti-metastatic potential of Indirubin. Taken together, these results suggest that Indirubin exerts potent anticancer effects on glioma cells and may prove essential in the management of glioma.

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MINERALIZATION OF TEETH ENAMEL AFTER ERUPTION

Wiadomości Lekarskie ◽

10.36740/wlek202106101 ◽

2021 ◽

Vol 74 (6) ◽

pp. 1297-1301

Author(s):

Oleksij P. Kostyrenko ◽

Nataliia I. Vynnyk ◽

Mykhailo M. Koptev ◽

Petro A. Hasiuk ◽

Maksym I. Skrypnyk ◽

...

Keyword(s):

Light And Electron Microscopy ◽

Permanent Tooth ◽

Postnatal Period ◽

Permanent Teeth ◽

Tooth Crown ◽

Electron Microscopic ◽

Cervical Portion ◽

Immunohistochemical Methods ◽

Protein Deposit ◽

Different Parts

The aim: The paper was aimed at the study of the processes of mineralization of the enamel of the permanent tooth after its eruption. Materials and methods: To study the structure of the enamel of permanent teeth has been carried out using light and electron microscopy. The study of the process of the development of the primordia of the permanent teeth involved 10 culled puppies of 30-40 days of age. Microscopic, electron microscopic, immunohistochemical methods of research have been used to study the processes of histogenesis. Results: The studies show that in the postnatal period, the formation of the crown, externally covered with cuticular epithelium, marks the formation of the primordium of the permanent tooth at the follicle stage. After eruption of a tooth, different parts of its crown have three individual structural and functional barriers to enamel biomineralization. The first one is provided by the cuticular epithelium of the pitted areas of the crown, which ensures filtering of the salivary fluid from the protein deposit in the form of a pellicle. The second barrier is defined on the lateral and cuspidate surfaces of the enamel, where the cuticle is erased or poorly expressed. The third structural and functional barrier of enamel biomineralization is located in the cervical portion of teeth of different classes. Conclusions: Different areas of the enamel in the tooth crown have specific filtration barriers, which can be distinguished as follows: pit-and-fissure-and-groove, cuspidateand-approximal, and cervical barriers. The cuticle is poorly expressed or totally absent on the cusps of the tooth crowns in contrast to pitted areas.

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Mitomycin C Induces Apoptosis in Rheumatoid Arthritis Fibroblast-Like Synoviocytes via a Mitochondrial-Mediated Pathway

Cellular Physiology and Biochemistry ◽

10.1159/000373938 ◽

2015 ◽

Vol 35 (3) ◽

pp. 1125-1136 ◽

Cited By ~ 21

Author(s):

Chuqi Yan ◽

Dechao Kong ◽

Dong Ge ◽

Yanming Zhang ◽

Xishan Zhang ◽

...

Keyword(s):

Rheumatoid Arthritis ◽

Cell Viability ◽

Mitomycin C ◽

Apoptotic Cell ◽

Chronic Inflammatory Disease ◽

Cell Counting ◽

Terminal Deoxynucleotidyl Transferase ◽

Control Treatment ◽

Induced Apoptosis ◽

Fibroblast Like Synoviocytes

Background/Aims: Rheumatoid arthritis (RA) is a systemic chronic inflammatory disease characterised by prominent synoviocyte hyperplasia and a potential imbalance between the growth and death of fibroblast-like synoviocytes (FLS). Mitomycin C (MMC) has previously been demonstrated to inhibit fibroblast proliferation and to induce fibroblast apoptosis. However, the effects of MMC on the proliferation and apoptosis of human RA FLS and the potential mechanisms underlying its effects remain unknown. Methods: Cell viability was determined using the Cell Counting Kit-8 assay. Apoptotic cell death was analysed via Annexin V-FITC/PI double staining and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labelling. The production of intracellular reactive oxygen species (ROS) was assessed via flow cytometry, and the changes in mitochondrial membrane potential (ΔΨm) were visualized based on JC-1 staining via fluorescence microscopy. The expression of apoptosis-related proteins was determined via Western blot. Results: Treatment with MMC significantly reduced cell viability and induced apoptosis in RA FLS. Furthermore, MMC exposure was found to stimulate the production of ROS and to disrupt the ΔΨm compared to the control treatment. Moreover, MMC increased the release of mitochondrial cytochrome c, the ratio of Bax/Bcl-2, the activation of caspase-9 and caspase-3, and the subsequent cleavage of poly(ADP-ribose) polymerase. Conclusion: Our findings suggest that MMC inhibits cell proliferation and induces apoptosis in RA FLS, and the mechanism underlying this MMC-induced apoptosis may involve a mitochondrial signalling pathway.

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HGF Converts ErbB2/Neu Epithelial Morphogenesis to Cell Invasion

Molecular Biology of the Cell ◽

10.1091/mbc.e04-07-0567 ◽

2005 ◽

Vol 16 (2) ◽

pp. 550-561 ◽

Cited By ~ 77

Author(s):

Hanane Khoury ◽

Monica A. Naujokas ◽

Dongmei Zuo ◽

Veena Sangwan ◽

Melanie M. Frigault ◽

...

Keyword(s):

Growth Factor ◽

Hepatocyte Growth Factor ◽

Cell Invasion ◽

Single Cells ◽

Epithelial Morphogenesis ◽

Cell Junctions ◽

Junction Protein ◽

Hepatocyte Growth ◽

E Cadherin ◽

Cell Cell

Activation of the hepatocyte growth factor receptor Met induces a morphogenic response and stimulates the formation of branching tubules by Madin-Darby canine kidney (MDCK) epithelial cells in three-dimensional cultures. A constitutively activated ErbB2/Neu receptor, NeuNT, promotes a similar invasive morphogenic program in MDCK cells. Because both receptors are expressed in breast epithelia, are associated with poor prognosis, and hepatocyte growth factor (HGF) is expressed in stroma, we examined the consequence of cooperation between these signals. We show that HGF disrupts NeuNT-induced epithelial morphogenesis, stimulating the breakdown of cell-cell junctions, dispersal, and invasion of single cells. This correlates with a decrease in junctional proteins claudin-1 and E-cadherin, in addition to the internalization of the tight junction protein ZO-1. HGF-induced invasion of NT-expressing cells is abrogated by pretreatment with a pharmacological inhibitor of the mitogen-activated protein kinase kinase (MEK) pathway, which restores E-cadherin and ZO-1 at cell-cell junctions, establishing the involvement of MEK-dependent pathways in this process. These results demonstrate that physiological signals downstream from the HGF/Met receptor synergize with ErbB2/Neu to enhance the malignant phenotype, promoting the breakdown of cell-cell junctions and enhanced cell invasion. This is particularly important for cancers where ErbB2/Neu is overexpressed and HGF is a physiological growth factor found in the stroma.

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Induction of cell death in prostate cancer cells by escopoletin, a promising treatment strategy

Bangladesh Journal of Pharmacology ◽

10.3329/bjp.v10i3.23095 ◽

2015 ◽

Vol 10 (3) ◽

pp. 500 ◽

Cited By ~ 1

Author(s):

Da Chen ◽

Xiao-Yi Zhang ◽

Fa-Zhu Zheng ◽

Hai-Tao Wang ◽

Jian-Liang Cai ◽

...

Keyword(s):

Prostate Cancer ◽

Cell Death ◽

Antioxidant Activities ◽

Apoptotic Cell ◽

Annexin V ◽

Cycle Arrest ◽

Anti Cancer ◽

Enhanced Expression ◽

Promising Treatment ◽

Du145 Cells

<p>Escopoletin, a phenolic compound belonging to anthocyanin family shows promising antioxidant activities. In the present study, anti-cancer effects of escopoletin treatment in DU145 cells were investigated. The sulphorhodamine-B staining and annexin V and propidium iodide were respectively used for the analysis of cell viability and death. The results revealed a significantly higher cytotoxicity by escopoletin that caused cell death in DU145 cells. Escopoletin treatment in DU145 cells markedly inhibited cell growth through non-apoptotic cell death and induced significant reactive oxygen species (ROS) production. It also induced G1 cell cycle arrest and cyclin D1 accumulation through the enhanced expression of p21. However, the effect of escopoletin on DU145 cells was reversed by pretreatment with glutathione antioxidant. This suggests that escopoletin induced generation of ROS is responsible for the increased cytotoxicity in DU145 cells. Thus, escopoletin exhibits potential therapeutic efficacy for the treatment of prostate cancer.</p><p> </p>

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