Hydrocortisone Increases the Vinblastine-Induced Chromosomal Damages in L929 Cells Investigated by the Micronucleus Assay on Cytokinesis-Blocked Binucleated Cells

Micronucleus assay is a test used to evaluate genotoxic damage in cells, which can be caused by various factors, like ionizing radiation. Interactions between radiation energies and DNA can cause breakage, leading to use chromosomal mutations or loss of genetic material, important events that could be induced in solid tumors to mitigate its expansion within human body. Melanoma has been described as a tumor with increased radio resistance. This work evaluated micronuclei percentages (%MN) in human melanoma cells (SK-MEL-37), irradiated by gamma radiation, with doses between 0 and 16Gy. Cell suspensions were irradiated in PBS by a 60Co source in doses between 0 and 16Gy, and incubated by 48h. Then cell membranes were lysed in the presence of SYTOX Green and EMA dyes, preserving nuclear membranes. Using this method, EMA-stained nuclei could be discriminated as those derived from dead cells, and SYTOX nuclei and micronuclei could be quantified. Micronuclei percentages were found to be proportional to dose, (R2 = 0.997). Only the highest dose (16Gy) could induce statistically significant increase of MN (p<0.0001), although cultures irradiated by 4, 8 and 16Gy showed significant increase of dead cell fractions. Calculation of the nuclei-to-beads ratio showed that 8 and 16Gy could reduce melanoma cell proliferation. Results showed that although cell death and loss of proliferative capacity could be observed on cultures irradiated at lower doses, genotoxic damage could be induced only on a higher dose. Resistance to radiation-induced genotoxicity could explain a relatively high radio resistance of melanoma tumors.

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Estimation of Genotoxic Effects from GSM Base Stations on Four Successive Generations (F1 F4) of Albino Mice, Mus Musculus Using RAPD Bands and Micronucleus Assay

Journal of Environment and Human ◽

10.15764/eh.2014.02001 ◽

2014 ◽

Vol 2014 (2) ◽

pp. 1-8

Author(s):

Aneyo Ayisat ◽

◽

Otitoloju Akeem ◽

Fowora Muinah ◽

◽

...

Keyword(s):

Mus Musculus ◽

Micronucleus Assay ◽

Base Stations ◽

Genotoxic Effects ◽

Albino Mice ◽

Successive Generations

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Rodent Bone Marrow Micronucleus Assay. Test Substance: Solvent Yellow 33 2-(2-Quinolyl)-1,3-indandione

10.21236/ada536686 ◽

2011 ◽

Author(s):

Devaki Sadhu

Keyword(s):

Bone Marrow ◽

Micronucleus Assay ◽

Test Substance ◽

Bone Marrow Micronucleus

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Biological and Toxicological Evaluation of N-(4methyl-phenyl)-4-methylphthalimide on Bone Cancer in Mice

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520619666190207130732 ◽

2019 ◽

Vol 19 (5) ◽

pp. 667-676

Author(s):

José R. Santin ◽

Gislaine F. da Silva ◽

Maria V.D. Pastor ◽

Milena F. Broering ◽

Roberta Nunes ◽

...

Keyword(s):

Breast Cancer ◽

Bone Metastasis ◽

In Silico ◽

Bone Matrix ◽

Micronucleus Assay ◽

Repeated Treatment ◽

Toxicological Evaluation ◽

Toxicological Analysis ◽

Breast Cancer Bone Metastasis

Background: It was recently demonstrated that the phthalimide N-(4-methyl-phenyl)-4- methylphthalimide (MPMPH-1) has important effects against acute and chronic pain in mice, with a mechanism of action correlated to adenylyl cyclase inhibition. Furthermore, it was also demonstrated that phthalimide derivatives presented antiproliferative and anti-tumor effects. Considering the literature data, the present study evaluated the effects of MPMPH-1 on breast cancer bone metastasis and correlated painful symptom, and provided additional toxicological information about the compound and its possible metabolites. Methods: In silico toxicological analysis was supported by in vitro and in vivo experiments to demonstrate the anti-tumor and anti-hypersensitivity effects of the compound. Results: The data obtained with the in silico toxicological analysis demonstrated that MPMPH-1 has mutagenic potential, with a low to moderate level of confidence. The mutagenicity potential was in vivo confirmed by micronucleus assay. MPMPH-1 treatments in the breast cancer bone metastasis model were able to prevent the osteoclastic resorption of bone matrix. Regarding cartilage, degradation was considerably reduced within the zoledronic acid group, while in MPMPH-1, chondrocyte multiplication was observed in random areas, suggesting bone regeneration. Additionally, the repeated treatment of mice with MPMPH-1 (10 mg/kg, i.p.), once a day for up to 36 days, significantly reduces the hypersensitivity in animals with breast cancer bone metastasis. Conclusion: Together, the data herein obtained show that MPMPH-1 is relatively safe, and significantly control the cancer growth, allied to the reduction in bone reabsorption and stimulation of bone and cartilage regeneration. MPMPH-1 effects may be linked, at least in part, to the ability of the compound to interfere with adenylylcyclase pathway activation.

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Glucosamine Protects Rat Bone Marrow Cells Against Cisplatin-induced Genotoxicity and Cytotoxicity

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520619666190704164126 ◽

2019 ◽

Vol 19 (14) ◽

pp. 1695-1702 ◽

Cited By ~ 1

Author(s):

Mohsen Cheki ◽

Salman Jafari ◽

Masoud Najafi ◽

Aziz Mahmoudzadeh

Keyword(s):

Bone Marrow ◽

Bone Marrow Cells ◽

Micronucleus Assay ◽

Ros Generation ◽

Polychromatic Erythrocytes ◽

Hematological Analysis ◽

Antagonistic Potential ◽

Harmful Effects ◽

Rat Bone ◽

Marrow Cells

Background and Objective: Glucosamine is a widely prescribed dietary supplement used in the treatment of osteoarthritis. In the present study, the chemoprotectant ability of glucosamine was evaluated against cisplatin-induced genotoxicity and cytotoxicity in rat bone marrow cells. Methods: Glucosamine was orally administrated to rats at doses of 75 and 150 mg/kg body weight for seven consecutive days. On the seventh day, the rats were treated with a single injection of cisplatin (5 mg/kg, i.p.) at 1h after the last oral administration. The cisplatin antagonistic potential of glucosamine was assessed by micronucleus assay, Reactive Oxygen Species (ROS) level analysis, hematological analysis, and flow cytometry. Results: Glucosamine administration to cisplatin-treated rats significantly decreased the frequencies of Micronucleated Polychromatic Erythrocytes (MnPCEs) and Micronucleated Normchromatic Erythrocytes (MnNCEs), and also increased PCE/(PCE+NCE) ratio in bone marrow cells. Furthermore, treatment of rats with glucosamine before cisplatin significantly inhibited apoptosis, necrosis and ROS generation in bone marrow cells, and also increased red blood cells count in peripheral blood. Conclusion: This study shows glucosamine to be a new effective chemoprotector against cisplatin-induced DNA damage and apoptosis in rat bone marrow cells. The results of this study may be helpful in reducing the harmful effects of cisplatin-based chemotherapy in the future.

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A Comparative Analysis of Methods for Titering Reovirus

Pharmaceutical Nanotechnology ◽

10.2174/2211738508666200826103322 ◽

2020 ◽

Vol 8 (5) ◽

pp. 409-417

Author(s):

Yi-Chen Yang ◽

Xian-Yao Wang ◽

Yuan-Yuan An ◽

Chun-Xiang Liao ◽

Nian-Xue Wang ◽

...

Keyword(s):

Traditional Method ◽

Virus Titer ◽

Cell Analysis ◽

Half Cell ◽

L929 Cells ◽

Cell Index ◽

Standard Curve ◽

Analysis Technique ◽

Curve Method ◽

Standard Curve Method

Background: A key challenge in the process of virus amplification is the need for a simple and convenient method for measuring virus titers. Objective: Real-time unlabeled cell analysis (RTCA) was used to establish a standard curve of correlation between half-cell index time (CIT50) and virus titer. At the same time, the virus titer from tunable resistance pulse detection (TRPS) technology was compared with the traditional median tissue culture infectious dose (TCID50) method to evaluate the feasibility and application value of the RTCA technique and TRPS technology. Methods: : Cell index (CI) values for L929 cells under different culture conditions were detected, and the appropriate initial cell inoculation density was screened. The half-cell index (CI50) values of reovirus infected L929 cells with TCID50 titers were analyzed by RTCA, the CI50-TCID50 standard curve was created, and a regression equation was developed. RTCA, TCID50, and TRPS methods were used to detect the reovirus titer obtained by the amplification, and the sensitivity and feasibility of the CIT50-TCID50 standard curve method were analyzed. The virus titer was detected by TRPS technology and the TCID50 method. Results: L929 cells were best propagated at an initial density of 6 × 103 cells/well. After infecting L929 cells with different titers of reference reovirus, the linear correlation of CIT50 and TCID50 was y = -2.1806x + 71.023 (R2 = 0.9742). The titer resulting from the RTCA assay was 7×109.6821 pfu/mL, from the TRPS assay was 4.52×1010 pfu/mL, and from the TCID50 assay was 7×109.467 pfu/mL. Conclusion: The CIT50-TCID50 standard curve method established by the RTCA technique can be used to quantitatively detect reovirus titer with L929 cells. Compared with the TCID50 method, it takes a relatively short time and has high sensitivity and accuracy. The TRPS technology requires even less time to quantify the virus, but its precision is lower than that of the TCID50 method and RTCA technology. This study provides new technical methods for assessing the virulence of infectious live reovirus particles. Lay Summary: After amplification of the virus, we need to detect the virus titers (the virulence of the virus). The traditional method is to use the virus to infect cells, and then the virus titers can be calculated by 50% of the cells infected. However, this traditional method is time consuming. The ways of RTCA (a real-time cell analysis technique) and TRPS (a nano-bioparticle analysis technique) help us to detect viral titers. The consistency of these three methods determines their feasibility and accuracy. If they are feasible, then these two simple technologies will provide new ideas for detecting viral titers.

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