hoechst 33258
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2021 ◽  
Vol 88 (6) ◽  
pp. 942-947
Author(s):  
A. P. Antonyan ◽  
N. H. Petrosyan ◽  
P. O. Vardevanyan

The comparative study on interaction of bisbenzimidazole compound Hoechst 33258 and thiazine dye methylene blue (MB) with bovine serum albumin (BSA) was carried out by spectroscopic methods. Denaturation curves as well as absorption spectra and differential absorption spectra of protein-ligand complexes were obtained. Denaturation temperature of albumin complexes of BSA with Hoechst 33258 was shown to decrease with the growth of concentration ratio of ligand/protein, while for MB, vice versa, denaturation temperature increases. Changes in absorption spectra and differential absorption spectra of the complexes of ligands with albumin were revealed, which result from the binding of these DNA-specific ligands to protein. It is supposed that at the interaction of Hoechst 33258 with BSA some loosening of protein compact structure occurs due to the partial loss of helicity of α-structures, while for MB an increase of the protein compact structure takes place.


2021 ◽  
Vol 55 (2 (255)) ◽  
pp. 158-164
Author(s):  
Nara H. Petrosyan

The study on the interaction of DNA-specific low-molecular compounds – groove binding material Hoechst 33258 and intercalating ligand methylene blue (MB) with serum albumin has been carried out. The absorption and differential absorption spectra of complexes of the mentioned ligands with protein were obtained. Changes of the absorption and differential absorption spectra indicate the binding of two ligands with albumin. The obtained results indicate that at the interaction with both ligands, the conformational state of the protein alters, though these changes are not similar, since in the case of MB a compactization of the protein folding occurs, while in the case of Hoechst 33258, most apparently, an unfolding of the compact structure takes place as a result of partial loss of helicity of $\alpha$-structures.


2021 ◽  
Vol 55 (2 (255)) ◽  
pp. 136-143
Author(s):  
Poghos H. Vardevanyan ◽  
Mariam A. Shahinyan ◽  
Anna V. Vardanyan ◽  
Svetlana V. Grigoryan

In this work the effect of millimeter range electromagnetic waves (MM EMW) with the frequency 64.5 GHz on the complexes of Hoechst 33258 (H33258) with DNA and human serum albumin (HSA) has been studied by the methods of absorption and fluorescence spectroscopies. It was shown that the irradiation results in weakening of H33258 interaction with both macromolecules, which is connected with the fact that the frequency 64.5 GHz, being resonant for water, leads to the structurizing of water component around DNA and HSA, due to which the binding becomes weaker. This conclusion is based on the values of both binding constants and Stern–Volmer constants.


2021 ◽  
Vol 55 (2 (255)) ◽  
pp. 144-150
Author(s):  
Marine A. Parsadanyan ◽  
Mariam A. Shahinyan ◽  
Zvart H. Movsisyan ◽  
Ara P. Antonyan

Study on the interaction of DNA-specific ligands – classical intercalator acridine orange (AO) and groove binding compound Hoechst 33258 (H33258) with poly(rA)-poly(rU), being a model for double-stranded (ds-) RNA, has been carried out. The absorption and fluorescence spectra of the complexes of these ligands with ds-polynucleotide were obtained. It was revealed that the optic and fluorescent characteristics of the complexes of both ligands with ds-RNA are similar with those at the complex-formation with DNA.


2021 ◽  
Vol 2021 ◽  
pp. 1-6
Author(s):  
Zeyu Xing ◽  
Xin Wang ◽  
Jiaqi Liu ◽  
Gang Liu ◽  
Menglu Zhang ◽  
...  

Purpose. To explore the effects of ulinastatin on the proliferation and apoptosis of breast cancer cells and the relevant mechanism of action. Methods. Breast cancer cells (MCF-7) were cultured and randomly divided into three groups, namely, control group, ulinastatin group, and ulinastatin+extracellular-regulated protein kinase (ERK) inhibitor group. Then, the Cell Counting Kit-8 (CCK-8) assay was carried out to detect the effect of ulinastatin on the viability of breast cancer cells. The effects of ulinastatin on the proliferation and apoptosis of breast cancer cells were determined via EdU staining and Hoechst 33258 staining assays, respectively. The messenger ribonucleic acid (mRNA) and protein expression levels of ERK and forkhead box O3 (FOXO3) in breast cancer cells were measured through reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. Results. In comparison with the control group, the ulinastatin group displayed decreased viability of breast cancer cells, a decreased positive rate of 5-ethynyl-2 ′ -deoxyuridine (EdU) staining, an increased positive rate of Hoechst 33258 staining, and reduced mRNA and protein levels of ERK and FOXO3 in breast cancer cells. Compared with those in the ulinastatin group, the viability of breast cancer cells was lowered, the positive rate of EdU staining was reduced, the positive rate of Hoechst 33258 staining was raised, and the mRNA and protein levels of ERK and FOXO3 in breast cancer cells clearly declined in the ulinastatin+ERK inhibitor group. Conclusion. Ulinastatin inhibits the proliferation and promotes the apoptosis of breast cancer cells. The possible mechanism of action is associated with the suppression of the ERK signaling pathway.


2021 ◽  
Vol 11 (7) ◽  
pp. 1346-1351
Author(s):  
Xiaowei Xie ◽  
Jiansheng Lin ◽  
Mingqiang Kang ◽  
Ying Guo

Our study investigated miR-222’s effect on NSCLC apoptosis and proliferation. H1650 cells were assigned into NSCLC group, SI group and IN group followed by analysis of Akt-mTOR protein content, cell apoptosis and proliferation, and miR-222 expression by Western blot, Hoechst 33258 fluorescence staining, qRT-PCR, cck-8, and immunohistochemistry miR-222 expression was lowest in IN group and highest in SI group (P < 0.05). The nucleus shrinkage rate in IN group and NSCLC group was significantly higher than that in SI group (P < 0.05) with NSCLC group showed more cells apoptosis than IN group (P < 0.05). The SI group had significantly higher OD value than IN group and NSCLC group (P < 0.05) with NSCLC group having significantly higher OD value than IN group (P < 0.05). The Akt-mTOR and p-Akt/p-mTOR expression was lowest in IN group and highest in SI group (P < 0.05) with lower level in IN group than NSCLC group (P < 0.05). The number of positive Akt-mTOR of H1650 cells was highest in SI group while lowest in IN group (P < 0.05). The decreased miR-222 expression in NSCLC can promote cancer cell apoptosis and inhibit lung cancer development, which may be via down-regulating Akt-mTOR signaling.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Majtnerova Pavlina ◽  
Capek Jan ◽  
Petira Filip ◽  
Handl Jiri ◽  
Rousar Tomas

AbstractAt present, nuclear condensation and fragmentation have been estimated also using Hoechst probes in fluorescence microscopy and flow cytometry. However, none of the methods used the Hoechst probes for quantitative spectrofluorometric assessment. Therefore, the aim of the present study was to develop a spectrofluorometric assay for detection of nuclear condensation and fragmentation in the intact cells. We used human hepatoma HepG2 and renal HK-2 cells cultured in 96-well plates treated with potent apoptotic inducers (i.e. cisplatin, staurosporine, camptothecin) for 6–48 h. Afterwards, the cells were incubated with Hoechst 33258 (2 µg/mL) and the increase of fluorescence after binding of the dye to DNA was measured. The developed spectrofluorometric assay was capable to detect nuclear changes caused by all tested apoptotic inducers. Then, we compared the outcomes of the spectrofluorometric assay with other methods detecting cell impairment and apoptosis (i.e. WST-1 and glutathione tests, TUNEL, DNA ladder, caspase activity, PARP-1 and JNKs expressions). We found that our developed spectrofluorometric assay provided results of the same sensitivity as the TUNEL assay but with the advantages of being fast processing, low-cost and a high throughput. Because nuclear condensation and fragmentation can be typical markers of cell death, especially in apoptosis, we suppose that the spectrofluorometric assay could become a routinely used method for characterizing cell death processes.


2021 ◽  
Vol 55 (1 (254)) ◽  
pp. 39-45
Author(s):  
Marine A. Parsadanyan

The study of complexes of groove binding ligand Hoechst 33258 (H33258) with Calf Thymus DNA has been carried out. The data obtained revealed that the melting curves of the complexes of H33258 with DNA are monophasic at low ligand concentrations (0 < r ≤ 0.2) and become biphasic at relatively high concentrations (0.2 < r ≤ 0.33). This effect was revealed to depend on the ionic strength of the solution, and can also occur at high concentrations of the ligand. Comparison of the obtained data with the results for poly(rA)-poly(rU) and poly(dA)-poly(dT) shows a coincidence in the case of DNA and poly(rA)-poly(rU), while in the case of poly(dA)-poly(dT) the melting curves become biphasic at low ligand concentrations and actually do not depend on the ionic strength of the solution.


2021 ◽  
Vol 5 (2) ◽  
Author(s):  
Yuming Zhang ◽  
Zhijun Mao ◽  
Fei Xue ◽  
Yu Zhang ◽  
Yuting Min ◽  
...  

Objective: To explore the effect of propofol (Prof) on the proliferation, migration and invasion of human gastric cancer cell MGC-803 and its molecular mechanism. Methods: The MTT method was used to study the effects of Prof with different doses and durations on the viability of MGC-803 cells. Hoechst 33258 staining and electron microscopy were used to detect the effects of Prof on MGC-803 cell apoptosis. Transwell experiments were used to detect the effects of Prof on the migration and invasion of MGC-803 cells. RT-PCR detects the effect of Prof on the expression of miR-195 in MGC-803 cells, and Western Blot detects the effect of Prof on the protein expression of JAK/STAT signaling pathway. Results: Compared with 0mg/ml Prof, 5mg/ml, 10mg/ml and 20mg/ml Prof treatment with 24h, 48h and 72h can significantly reduce cell viability (P <0.05). Compared with the Control group, the percentage of Hoechst 33258 staining positive cells in the Prof group and the apoptosis rate under the electron microscope were significantly increased (P <0.05). Compared with the Control group, the cell migration rate and invasion rate of the Prof group were significantly reduced (P <0.05). Compared with the Control group, the expression of miRNA-195 in the Prof group cells was increased significantly (P <0.05). Compared with the Control group, the activity of p-Jak1 and p-STAT3 proteins in the Prof group were significantly reduced (P <0.05). Conclusion: Prof can reduce the cell viability, migration and invasion of gastric cancer cell MGC-803, and promote its apoptosis. Its mechanism may be related to the promotion of miR-195 expression and inhibition of JAK/STAT signal pathway activity.


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