Comet assay and flow cytometry analysis of DNA repair in normal and cancer cells treated with known mutagens with different mechanisms of action

2003 ◽  
Vol 23 (S2) ◽  
pp. 13-29 ◽  
Author(s):  
M. Suggitt ◽  
J. Fearnley ◽  
D.J. Swaine ◽  
M. Volpato ◽  
R.M. Phillips ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Dna Repair ◽  
Comet Assay ◽  
Cancer Cells ◽  
Mechanisms Of Action ◽  
Flow Cytometry Analysis

scholarly journals Cisplatin Increased the Expression of PD-1 and PD-L1 by YAP1 in Hepatocellular Carcinoma

2021 ◽  
Author(s):  
Liyuan Hao ◽  
Yinglin Guo ◽  
Qing Peng ◽  
Zhiqin Zhang ◽  
Shenghao Li ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Flow Cytometry ◽  
T Cells ◽  
Cancer Cells ◽  
Flow Cytometry Analysis ◽  
Chemotherapy Drugs ◽  
The World ◽  
Hcc Cells

Abstract Hepatocellular carcinoma (HCC) was one of the most malignant cancers in the world. Cisplatin (DDP) was one of the main chemotherapy drugs for HCC, but the mechanism of DDP treatment for HCC remains unclear. In this presentation, we found that DDP inhibited the growth of HCC cells and promoted the expression of PD-1 and its ligand PD-L1 in cancer cells. Meanwhile, flow cytometry analysis revealed that DDP enhanced PD-1-CD8+ T cells expression and decreased PD-1+CD8+ T cells expression. ELISA analysis suggested that DDP decreased TGF-β expression in vivo. Therefore, the study indicated that DDP enhanced PD-1 and PD-L1 expression and inhibited the growth of HCC.

Effect of heterogeneity of HER2 expression on brain metastases of breast cancer.

2012 ◽  
Vol 30 (15_suppl) ◽  
pp. 635-635
Author(s):  
Mari Hosonaga ◽  
Yoshimi Arima ◽  
Eiji Sugihara ◽  
Norio Kohno ◽  
Hideyuki Saya
Keyword(s):  
Breast Cancer ◽  
Flow Cytometry ◽  
Brain Metastases ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Triple Negative ◽  
Her2 Expression ◽  
Flow Cytometry Analysis ◽  
Her2 Positive ◽  
The Brain

635 Background: HER2-overexpressing or triple-negative [ER(-)/PR(-)/HER2(-)] breast cancers are associated with increased risk of brain metastases. The mechanisms leading to metastasis in each subtype are not well known. Methods: We introduced the wild-type HER2 gene into MDA-MB-231-luc-D3H2LN (231-Luc) cells, which are triple-negative breast cancer cells, and established HER2-expressing (63.2%) cells as 231-Luc-HER2 cells. We investigated the tumor formation following orthotopic inoculation and brain metastasis following intracardiac injection into nude mice. Metastasis was detected by bioluminescence imaging and confirmed in H&E staining and immunohistochemistry of vimentin and HER2 expressions. Flow cytometry analysis was used to detect the proportion of CD44+/CD24- cells, a maker for stem-like breast cancer cells. Results: 231-Luc-HER2 cells formed larger tumors in orthotopic xenograft models compared to 231-Luc cells, however, no significant difference was observed in proliferation in vitro. Neither 231-Luc-HER2 nor 231-Luc metastasized in the brain from the breast after orthotopic inoculation. After intracardiac injections of the 231-Luc-HER2 cells, brain metastasis developed (7/13 mice, 53.8%). Immunohistochemical analysis revealed that most metastasized cells expressed HER2, although we had injected a mixture of HER2-positive and HER2-negative cancer cells. Interestingly, administering Lapatinib, a dual EGFR and HER2 tyrosine kinase inhibitor, effectively prevented HER2-positive cells to colonize the brain. However, the HER2-negative 231-Luc-HER2 cells developed into brain metastases. In fact, the 231-Luc cells, which are HER2-negative, also metastasized in the brain (10/16 mice, 62.5%). Flow cytometry analysis of the 231-Luc-HER2 cells showed that HER2-positive cells decreased the population of CD44+/CD24- (HER2+/CD44+/CD24-: 86.8% and HER2-/CD44+/CD24-: 96.3%). Conclusions: The mechanism of brain metastases of HER2-positive breast cancer cells is different from that of HER2-negative breast cancer cells. It is therefore important to consider an additional therapeutic approach when dealing with HER2-negative cells in tumors having the heterogeneity of HER2 expression.

scholarly journals Phospholipase A2 from krait Bungarus fasciatus venom induces human cancer cell death in vitro

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e8055 ◽  
Author(s):  
Thien V. Tran ◽  
Andrei E. Siniavin ◽  
Anh N. Hoang ◽  
My T.T. Le ◽  
Chuong D. Pham ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cell Death ◽  
Cancer Cells ◽  
Cell Lines ◽  
Phospholipase A ◽  
Human Lung Cancer ◽  
Flow Cytometry Analysis ◽  
Mcf7 Cells ◽  
Ki 67 ◽  
Cellular Marker

Background Snake venoms are the complex mixtures of different compounds manifesting a wide array of biological activities. The venoms of kraits (genus Bungarus, family Elapidae) induce mainly neurological symptoms; however, these venoms show a cytotoxicity against cancer cells as well. This study was conducted to identify in Bungarus fasciatus venom an active compound(s) exerting cytotoxic effects toward MCF7 human breast cancer cells and A549 human lung cancer cells. Methods The crude venom of B. fasciatus was separated by gel-filtration on Superdex HR 75 column and reversed phase HPLC on C18 column. The fractions obtained were screened for cytotoxic effect against MCF7, A549, and HK2 cell lines using colorimetric assay with the tetrazolium dye MTT- 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. The primary structure of active protein was established by ultra high resolution LC-MS/MS. The molecular mechanism of the isolated protein action on MCF7 cells was elucidated by flow cytometry. Results MTT cell viability assays of cancer cells incubated with fractions isolated from B. fasciatus venom revealed a protein with molecular mass of about 13 kDa possessing significant cytotoxicity. This protein manifested the dose and time dependent cytotoxicity for MCF7 and A549 cell lines while showed no toxic effect on human normal kidney HK2 cells. In MCF7, flow cytometry analysis revealed a decrease in the proportion of Ki-67 positive cells. As Ki-67 protein is a cellular marker for proliferation, its decline indicates the reduction in the proliferation of MCF7 cells treated with the protein. Flow cytometry analysis of MCF7 cells stained with propidium iodide and Annexin V conjugated with allophycocyanin showed that a probable mechanism of cell death is apoptosis. Mass spectrometric studies showed that the cytotoxic protein was phospholipase A2. The amino acid sequence of this enzyme earlier was deduced from cloned cDNA, and in this work it was isolated from the venom as a protein for the first time. It is also the first krait phospholipase A2 manifesting the cytotoxicity for cancer cells.

Comet assay and DNA flow cytometry analysis of staurosporine-induced apoptosis

Cytometry ◽  
1999 ◽  
Vol 36 (2) ◽  
pp. 117-122 ◽  
Author(s):  
Thierry Godard ◽  
Edwige Deslandes ◽  
Pierre Lebailly ◽  
Carole Vigreux ◽  
Laurent Poulain ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Comet Assay ◽  
Dna Flow Cytometry ◽  
Flow Cytometry Analysis ◽  
Induced Apoptosis

Nickel(II) Affects Poly(ADP-ribose) Polymerase-Mediated DNA Repair in Normal and Cancer Cells

2006 ◽  
Vol 61 (1-2) ◽  
pp. 142-148 ◽  
Author(s):  
Katarzyna Wozniak ◽  
Agnieszka Czechowska ◽  
Janusz Blasiak
Keyword(s):  
Dna Repair ◽  
Comet Assay ◽  
Cancer Cells ◽  
Nickel Chloride ◽  
Repair Process ◽  
Alkaline Comet Assay ◽  
Γ Radiation ◽  
Potential Target

Abstract Nickel(II) can be genotoxic, but the mechanism of its genotoxicity is not fully understood and the process of DNA repair may be considered as its potential target. We studied the effect of nickel chloride on the poly(ADP-ribose) polymerase (PARP)-mediated repair of DNA damaged by γ-radiation and idarubicin with the alkaline comet assay in normal and cancer cells. Our results indicate that nickel chloride at very low, non-cytotoxic concentration of 1 μᴍ can affect PARP-mediated DNA repair of lesions evoked by idarubicin and γ-radiation. We also suggest that in the quiescent lymphocytes treated with γ-radiation, nickel(II) could interfere with DNA repair process independent of PARP

scholarly journals Production of monoclonal antibody to quantify serum CA125 concentration in breast and ovarian cancer patients

2020 ◽  
Vol 42 (4) ◽  
Author(s):  
Nguyen Thi Xuan ◽  
Nguyen Trong Ha
Keyword(s):  
Flow Cytometry ◽  
Cancer Cells ◽  
Sandwich Elisa ◽  
Flow Cytometry Analysis ◽  
Brca1 And Brca2 ◽  
Elisa System ◽  
Serum Ca125 ◽  
Capture Antibody ◽  
Hybridoma Cell Lines ◽  
Tumor Growth And Metastasis

Ovarian carcinoma (OC) and breast cancer (BC) are mainly caused by alterations in genes such as BRCA1 and BRCA2, which are involved in differentiation and survival of cancer cells. The protein CA125 (MUC16) is released by cancer cells in most OC and a few BC patients. The cut-off point of CA125 level for tumor growth and metastasis is 35 U/mL. Production of CA125 monoclonal antidody (mAb) to determine expression level of this antigen by ELISA has been well known. In this study, we aimed to generate CA125-specific mAb for developing a new in-house ELISA kit. To this end, BALB/c mice were immunized with CA125 protein and splenocytes of immunized mice were fused with myeloma Sp2/0 cells. Efficiency of mAbs secreted from the hybridoma clones was examined by ELISA and flow cytometry analysis. As a result, among 3 stable hybridoma cell lines identified, A1 clone attained about 90% positive for anti-CA125 mAb, whereas H1 and H3 clones were about 40% and 50% positive for anti-CA125 mAb, respectively. By flow cytometry analysis, anti-CA125 mAb from A1 clone was more specific to CA125 antigen present in OVCAR -3 cells than those from H1 or H3 clone. In addition, the isotype of the obtained mAb was specific IgG1 and Kappa light chain. In conclusion, the mouse anti-human CA125 mAb generated in our lab was specifically binding to CA125 antigen and used as the capture antibody in sandwich ELISA system for early diagnosis as well as monitoring therapeutic response in OC patients.  

scholarly journals Nuclear body phase separation drives telomere clustering in ALT cancer cells

2020 ◽  
Vol 31 (18) ◽  
pp. 2048-2056 ◽  
Author(s):  
Huaiying Zhang ◽  
Rongwei Zhao ◽  
Jason Tones ◽  
Michel Liu ◽  
Robert L. Dilley ◽  
...  
Keyword(s):  
Dna Repair ◽  
Phase Separation ◽  
Cancer Cells ◽  
Nuclear Body ◽  
Promyelocytic Leukemia ◽  
Nuclear Bodies ◽  
Telomere Elongation ◽  
Alternative Lengthening ◽  
Induce Phase Separation ◽  
Induce Phase

A chemical dimerization approach is developed to induce phase separation of APB nuclear bodies involved in telomere elongation in alternative lengthening of telomeres (ALT) cancer cells. It reveals that ALT telomere-associated promyelocytic leukemia nuclear body (APB) fusion leads to telomere clustering to provide templates for homology-directed telomere synthesis, an ability that is decoupled from APB function in enriching DNA repair factors.

Pharmacological targeting of differential DNA repair, radio-sensitizes WRN-deficient cancer cells in vitro and in vivo

2021 ◽  
Vol 186 ◽  
pp. 114450
Author(s):  
Pooja Gupta ◽  
Bhaskar Saha ◽  
Subrata Chattopadhyay ◽  
Birija Sankar Patro
Keyword(s):  
Dna Repair ◽  
Cancer Cells
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