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بخشی از مقاله انگلیسیعنوان انگلیسی:Determining the interaction behavior of calf thymus DNA with berberine hydrochloride in the presence of linker histone: a biophysical study~~en~~

Abstract

The binding of small molecules with histone-DNA complexes can cause an interference in vital cellular processes such as cell division and the growth of cancerous cells that results in apoptosis. It is significant to study the interaction of small molecules with histone-DNA complex for the purpose of better understanding their mechanism of action, as well as designing novel and more effective drug compounds. The fluorescence quenching of ct-DNA upon interaction with Berberine has determined the binding of Berberine to ct-DNA with Ksv= 9.46 107 M1 . Ksv value of ct-DNA-Berberine in the presence of H1 has been observed to be 3.10 107 M1 , indicating that the H1 has caused a reduction in the binding affinity of Berberine to ct-DNA. In the competitive emission spectrum, ethidium bromide (EB) and acridine orange (AO) have been examined as intercalators through the addition of Berberine to ct-DNA complexes, which includes ctDNA-EB and ctDNA-AO. Although in the presence of histone H1 , we have observed signs of competition through the induced changes within the emission spectra, yet there has been apparently no competition between the ligands and probes. The viscosity results have confirmed the different behaviors of interaction between ctDNA and Berberine throughout the binary and ternary systems. We have figured out the IC50 and viability percent values at three different time durations of interaction between Berberine and MCF7 cell line. The molecular experiments have been completed by achieving the results of MTT assay, which have been confirmed to be in good agreement with molecular modeling studies.

Introduction

Cancer is acknowledged as the collapse of cellular regulation and uncontrolled cell division. DNA carries unique genetic information about all the living creatures and controls cellular regulations, which acts as the guiding agent for the biological procedures of cells that result in transcription replication and protein synthesis (Agarwal, Jangir, & Mehrotra, 2013; Anbazhagan & Renganathan, 2008; Arrondo, Muga, Castresana, & Goni, ~ 1993; Asadi, Safaei, Ranjbar, & Hasani, 2004). Cell DNA is known as the compact form of chromatin, and among the group of proteins called histones, which are involved in the formation of chromatin, histone H1 is labeled as their most important member. Hence, the DNA and histone–DNA complex have the potential of being utilized as therapeutic goals for designing anticancer drugs (Bazett-Jones, Cot ^ e, Landel, Peterson, & Workman, 1999).

Generally, there are three different ways that the anticancer drugs can interact with DNA including binding to DNA groove (groove binding interaction), the binding between two strands of DNA (intercalation interaction) and covalent binding. Drugs with intercalation interactions can eliminate the cancer cells by creating a binding between two DNA strands (Bera, Sahoo, Ghosh, & Dasgupta, 2008; Bi et al., 2006; Blake et al., 1999; Borchman, Yappert, & Herrell, 1991; Burton et al., 1978; Cao & He, 1998; Chen et al., 2005; Chi & Liu, 2011; Du et al., 2011; Sowrirajan, Yousuf, & Enoch, 2014). We have investigated the interaction of Berberine chloride with DNA and histone H1–DNA complex throughout this study. Berberine (C20H18NO4) is a chemical compound with the molar mass of 366.36122 g/mol (Figure 1) (Gallori et al., 2000; Gittelson & Walker, 1967; Guo, Yue, & Gao, 2011; Hossain & Kumar, 2009; Huang & Zou, 2006; Ivankin, Carson, Kinney, & Wanunu, 2013; Jordan & Wilson, 2004; Kashanian et al., 2008; Khare & Pande, 2012; Kumar, Naik, Girija, Sharath, & Pradeepa, 2012; Kypr & Vorlckova, 2002; Lerman, 1961). It has been widely studied as an isokoinoline alkaloid that belongs to the Protobebrine structural class and contains a cholesterol-like construction. In accordance with the available data, Berberine is capable of various biological and clinical activities such as antimicrobial, anticancer, antiinflammatory and antioxidant effects (Li & Dong, 2009; Macquet & Butour, 1978; Nafisi, Saboury, Keramat, Neault, & Tajmir-Riahi, 2007; Neelam, Gokara, Sudhamalla, Amooru, & Subramanyam, 2010). This particular substance can induce apoptosis through NFKB pathway and inhibit cell proliferation, as well as prevent and cause a delay in the progress of angiogenesis (Oohara & Wada, 1987). In addition, Jin et al. have studied the outcomes of Berberine on SKOV3 cancer cell line and reported that it has shown antitumor effects (Jin, Zhang, & Li, 2015). Berberine is also carried out to inhibit cancer cell proliferation by reducing the expression of BCL2 anti-apoptotic genes and increasing the expression of proapoptotic genes. Moreover, this interesting substance can induce apoptosis in the MCF-7 breast cancer cell line by causing a decrease in the expression of BCL2 and increasing the expression of CytC genes. Berberine has been widely studied and proven to contain antioxidant, antibacterial, antiTB and antitumor properties (Tang et al., 2009).

In this research, we have performed investigations on the type of interaction between Berberine chloride with DNA and histone H1–DNA complex throughout a variety of methods including fluorescence spectroscopy, absorption spectroscopy, resonance light scattering, circular dichroism (CD), melting temperature measurement, viscosity measurements, molecular modeling and 3-(4,5-cimethylthiazol-2-yl)-2,5- diphenyl tetrazolium bromide (MTT) assay

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