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Fenugreek in Management of Immunological, Infectious, and Malignant Disorders
Published in Dilip Ghosh, Prasad Thakurdesai, Fenugreek, 2022
Rohini Pujari, Prasad Thakurdesai
The exposure of the methanolic extract of fenugreek seeds (100 to 500 ??g/mL) to HepG2 cell line is reported to result in dose-dependent caspase-mediated apoptosis as evidenced by MTT assay, cell morphology alterations, enzyme-linked immunosorbent assay (ELISA), flow cytometric analysis, caspase-3 activity, and expression of p53, a proapoptotic protein, Bax, and proliferating cell nuclear antigen (PCNA) (Khalil et al. 2015). Inactivation of the p53 tumor suppressor is a frequent event in tumorigenesis, and apoptosis, including hepatocellular carcinoma. The p53 gene is documented to play a crucial role as a transcriptional activator in inducing the transcription of several genes, including apoptosis-related genes (Hussain et al. 2007; Rivlin et al. 2011). The Bax gene is a proapoptotic gene that acts as a transcriptional target of p53 (Benchimol 2001; Moxley and Reisman 2021). PCNA is responsible for several cellular mechanisms, including apoptosis, cell cycle regulation, DNA synthesis and repair, and interaction with various proteins in a p53-dependent manner (Chen 2016). Another study reported the dose-dependent cytotoxicity efficacy of ethanolic extract of fenugreek in a reduction in cell viability, increasing the cell mortality of HepG2 cell lines during in vitro MTT assay, perhaps through antioxidant properties (Al-Dabbagh et al. 2018).
Flow Cytometric Analysis of Human Bone Marrow
Published in Adrian P. Gee, BONE MARROW PROCESSING and PURGING, 2020
James G. Bender, Dennis Van Epps
An alternative method for identifying the proliferating fraction of cells in the marrow is to use antibodies to nuclear proteins associated with proliferation. These proteins include the proliferating cell nuclear antigen (PCNA), also called cyclin, which is an auxiliary factor for DNA polymerase delta,74,80 and a nuclear protein identified by the Ki-67 antibody expressed in cycling cells, including the G1 phase of the daughter population.73,75 Expression of the protein identified by Ki-67 therefore identifies a slightly larger population of proliferating cells than DNA stains (PI).77 Multiparameter analysis of both Ki-67 and PCNA has been used to resolve distinct G1, S, G2, and M populations of proliferating cell lines.74 The use of Ki-67 to detect proliferating cells is adaptable for multicolor analysis, as it can be used with any fluorochrome, and the complete resolution of the proliferating population eliminates the problems of calculating the cells in early S-phase using mathematical algorithms.77 Although Ki-67 and antibodies to PNCA have been used for the evaluation of malignant cells, including those in bone marrow,76 an evaluation of the proliferating fraction of normal cells in normal marrow has not been made.
Synthetic DNA-Based Compounds for the Prevention of Coronary Restenosis: Current Status and Future Challenges
Published in Eric Wickstrom, Clinical Trials of Genetic Therapy with Antisense DNA and DNA Vectors, 2020
Andrew Zalewski, Yi Shi, John D. Mannion, Femando Roqué
PCNA is a cofactor of DNA polymerase, both of which are required for DNA replication (Prelich and Stillman, 1988). The antiproliferative effect of PCNA antisense (phosphorothioate) in combination with oligonucleotides directed against cell division cycle 2 (cdc2) kinase has been demonstrated. This represents an example of combined oligomer application against multiple targets to enhance antisense effects and to reduce nonspecific cell inhibition (Morishita et al., 1993). Antisense targeting PCNA alone has also been demonstrated to reduce PCNA expression and neointimal formation in a rat model after periadventitial delivery in a pluronic gel (Simons et al., 1994).
Exogenous glutathione protects against gentamicin-induced acute kidney injury by inhibiting NF-κB pathway, oxidative stress, and apoptosis and regulating PCNA
Published in Drug and Chemical Toxicology, 2023
Esmaeel Babaeenezhad, Omid Dezfoulian, Forouzan Hadipour Moradi, Sobhan Rahimi Monfared, Mohammad Davood Fattahi, Maryam Nasri, Abdolhakim Amini, Hassan Ahmadvand
PCNA has been established to contribute to DNA repair and replication (Tillhon et al. 2013). In an earlier study, Potocnjak et al. (Potočnjak and Domitrović 2016) disclosed that cisplatin as a nephrotoxic drug promoted the expression of P53, P21, and PCNA representing cell cycle arrest and DNA repair. Similarly, GM has been reported to upregulate P53 and P21 in renal proximal cells which could lead to cell cycle arrest (Denamur et al. 2016). In addition, like our results, several studies indicated that the expression of PCNA was elevated in the kidney after GM injection in animals (Bledsoe et al. 2008, Salama et al. 2018, Babaeenezhad et al. 2021). Accordingly, increased PCNA expression in GM-intoxicated rats may be related to cell cycle arrest and DNA repair in response to DNA injury resulting from GM intoxication (Choi et al. 2000). On the contrary, EGSH administration considerably reduced renal PCNA expression compared with animals received only GM which was in line with former researches (Bledsoe et al. 2008, Salama et al. 2018, Babaeenezhad et al. 2021). We speculated that decreased PCNA expression in the treated group was likely due to reducing DNA injury by EGSH (Kwon et al. 2019).
Fucoxanthin Inactivates the PI3K/Akt Signaling Pathway to Mediate Malignant Biological Behaviors of Non-Small Cell Lung Cancer
Published in Nutrition and Cancer, 2022
Xuehong Fang, Yuzhen Zhu, Taomin Zhang, Qian Li, Lvhua Fan, Xiaodan Li, Daishun Jiang, Jie Lin, Liyi Zou, Jianwei Ren, Zunnan Huang, Hua Ye, Yi Liu
To assess the effect of FX on cell proliferation, we first performed an MTT assay to detect the viability of NCI-H1299 and A549 cells following FX treatment. As shown in Figure 1A, B, FX could significantly inhibit the proliferation of NCI-H1299 and A549 cells in a time- and dose-dependent manner (P < 0.05) and IC50 of FX for NCI-H1299 and A549 cells treated for 24, 48, and 72 h were 41.41 ± 1.389 µM, 29.95 ± 2.791 µM, 22.79 ± 2.616 µM, and 30.34 ± 1.161 µM, 22.18 ± 2.449 µM, 17.66 ± 1.573 µM, respectively (Supplementary Table 1). Similar results were obtained by the colony formation assay (Figure 1C, D). Furthermore, we examined the expression of proliferating cell nuclear antigen (PCNA) in NCI-H1299 and A549 cells following treatment with FX for 24 h. PCNA participates in a variety of DNA metabolism processes, and targeting PCNA is an effective strategy to inhibit tumor cell proliferation (28). The different concentrations of FX significantly decreased the expression level of PCNA compared to the control group (Figure 1E, F). These results suggest that FX can effectively inhibit NSCLC cell proliferation.
Balanitoside as a Natural Adjuvant to Gemcitabine in Lung Cancer Experimental Model
Published in Nutrition and Cancer, 2022
Elsayed I. Salim, Sara S. Aboueisha, Abeer A. Khamis
Proliferating cell nuclear antigen (PCNA) is a well-conserved protein found in all eukaryotic species, as well as Archaea. PCNA was first discovered to be involved in DNA replication. PCNA function also participates in other cellular processes including chromatin remodeling, DNA repair, sister-chromatid cohesion, and cell cycle control. PCNA is a cell proliferation marker that is expressed in a variety of malignancies. Sections (4 µm), two from each lung lobe, and sections from all visible nodules or tumors morphologically appearing on the outer surface of the lungs were used for IHC. Briefly, sections were dewaxed, hydrated, rinsed in sodium citrate buffer (pH 6.0), and boiled for antigen retrieval. Sections were then treated with 0.3% hydrogen peroxide, normal horse serum, and anti-PCNA rabbit polyclonal antibody (Sigma-Aldrich, Cairo, Egypt) at a 1:500 dilution. After washing with Tris buffer solution, sections were developed using ABC-peroxidase reagents (Vector Laboratories, USA). As a negative control, normal serum was used instead of primary antibodies. IHC reactions were visualized using 3,3′-diaminobenzidine tetrahydrochloride and then counterstained with Mayer’s hematoxylin. The number of positively stained nuclei from high-power fields of the lung sections was counted and divided by the total number of nuclei × 100 to determine the PCNA labeling index (PCNA LI %).