Michigan Cancer Foundation-7 Cell Line Research Articles

The Efficacy of siRNA Specific MDM2 in the Induction of Apoptosis in MCF-7 Breast Cancer Cell Line

Background: A high number of human breast cancers overexpress the murine double minute (MDM2) gene which blocks the p53 protein which plays an important role in arresting the cell growth. The present study aimed to investigate the efficacy of siRNA specific MDM2 in knocking down MDM2 and its subsequent effects on p53 to exert antiproliferative effects on Michigan Cancer Foundation-7 (MCF-7) breast cancer cells. Method: In this in vitro study, we used the specific siRNA of the MDM2 gene to knock down the expression of the MDM2 protein in the MCF-7 cell line. The expression of MDM2, BCL2-associated X (BAX), BH3 interacting-domain death agonist (BID), and B cell lymphoma 2 (BCL2) genes was evaluated using the Real-time PCR technique. The apoptosis level was also assessed using the flow cytometry technique by the Annexin V test. Results: The results showed that the entry of MDM2 siRNA into MCF-7 cells significantly reduced the mRNA expression of MDM2 gene (P-value <0.05). Besides, the expression of the anti-apoptotic gene of BCL2 significantly decreased (P-value <0.05) in transfected MCF-7 cells, while that of BAX and BID genes increased (P-value <0.05). Conclusion: Based on the results, MDM2 inhibition is conducive to prevent cancer metastasis by the induction of cancer cell apoptosis. Moreover, it can be considered in cancer therapy along with chemotherapy.

Antimicrobial and cytotoxic activities of isoniazid connected menthone derivatives and their investigation of clinical pathogens causing infectious disease

BackgroundThis work is development of new molecules of isoniazid derivatives as dealing with potential of antimicrobial activity against clinical pathogens causing infectious disease. Antimicrobial of novel Mannich base derivatives can be achieved via one-pot synthesis in green chemistry approach. This method offers efficient, mild reaction conditions and high yields. In this study, totally 12 compounds (1a–l) was prepared and screened for cytotoxic and antimicrobial activities. Materials and methodsNewly synthesised compounds were conformed via FT- IR, 1H, and 13C NMR (Nuclear Magnetic Resonance), and mass spectra analysis. All compounds were checked antibacterial activity against gram-positive bacteria of Enterococcus faecalis, Staphylococcus aureus and gram-negative bacteria of Pseudomonas aeruginosa, Klebsiella pneumoniae and Escherichia coli. All compounds were checked against antifungal activity against Aspergillus fumigatus, Candida albicans, Cryptococcus neoformans, Aspergillus niger, and Microsporum audouinii. All compounds were screened for cytotoxic activity against the MCF-7 (Michigan Cancer Foundation-7) cancer cell line. ResultThe compound 1g was highly (MIC: 0.25 μg/mL) active against gram-negative bacterial of P. aeruginosa, whereas other compounds 1e and 1h were more active (MIC: 2 μg/mL) in K. pneumoniae and also 1g (MIC: 2 μg/mL) was more active in E. faecalis than standard ciprofloxacin. Antifungal screening, the compound 1b was highly active (MIC: 0.25 μg/mL) against C. albicance,1g (MIC: 2 μg/mL) and 1h (MIC: 4 μg/mL) was significant of active against A. fumigatus, and the compound 1c (MIC: 4 μg/mL) was extremely active in M. audouinii than clotrimazole. Compound 1g (GI50 = 0.01 μM) exhibited high activity against the MCF-7 cell line, while 1b (GI50 = 0.02 μM) was equipotent active compared with standard doxorubicin. ConclusionA novel set of isoniazid derivatives (1a–l) and 1h were synthesized and screened for antimicrobial and cytotoxic activities. We found some highly active molecules, which are evidencing to be a potential treatment of bacterial and fungal infection candidates.

Role of Amygdalin in Blocking DNA Replication in Breast Cancer In Vitro.

Amygdalin has anticancer benefits because of its active component, hydrocyanic acid. However, the underlying molecular mechanism is unclear. This study aimed to investigate the molecular mechanism by which amygdalin exerts antiproliferative effects in the human Michigan Cancer Foundation-7 (MCF-7) breast cancer cell line. MCF-7 cells were exposed to amygdalin at a particular IC50 value for 24 and 48 hours and compared to non-treated cells. An Affymetrix whole-transcript expression array was used to analyze the expression of 32 genes related to DNA replication. Among the 32 genes, amygdalin downregulated the expression of 16 genes and 19 genes by >1.5-fold at 24 and 48 hours, respectively. At 24 hours, the downregulated genes from the DNA polymerase α-primase complex were POLA1, POLA2, PRIM1, and PRIM2; DNA polymerase δ complex: POLD3; DNA polymerase ε complex: POLE4, Minichromosome Maintenance protein (MCM) complex (helicase): MCM2, MCM3, MCM4, MCM6, and MCM7; clamp and clamp loader: PCNA; nuclease: FEN1; and DNA ligase: LIG1. At 48 hours, the downregulated genes from the DNA polymerase α-primase complex were POLA1, POLA2, and PRIM1; DNA polymerase δ complex: POLD3; DNA polymerase ε complex: POLE and POLE2; MCM complex (helicase): MCM2, MCM3, MCM4, MCM5, MCM6, and MCM7; clamp and clamp loader: PCNA, RFC2, and RFC3; RNase H: RNASEH2A; nucleases: DNA2 and FEN1; and DNA ligase: LIG1. Amygdalin treatment caused downregulation of several genes that play critical roles in DNA replication in the MCF-7 cell line. Thus, it might be useful as an anticancer agent.

Study on Lidocaine Inhibiting Proliferation, Migration and Invasion of Breast Cancer Cells by Modulating the MicroRNA-495-3p/Fibroblast Growth Factor 9 Axis

To investigate the mechanism of lidocaine in inhibiting proliferation, migration and invasion of breast cancer cells. Human breast cancer cell line Michigan Cancer Foundation-7 cells were treated with lidocaine. Cell proliferation was detected by 2,5-diphenyl-2H-tetrazolium bromide, cell migration and invasion was detected by Transwell, microRNA-495-3p expression was detected by quantitative reverse transcription polymerase chain reaction and fibroblast growth factor 9, cyclin D1, matrix metalloproteinase 2 and matrix metalloproteinase 9 protein expression were detected by western blot. Human breast cancer cell line Michigan Cancer Foundation-7 cells were transfected with microRNA-495-3p inhibitor or fibroblast growth factor 9 overexpression vector. The effects of silencing microRNA-495-3p or fibroblast growth factor 9 overexpression on proliferation, migration and invasion of human breast cancer cell line Michigan Cancer Foundation-7 cells and the expression of cyclin D1, matrix metalloproteinase 2 and matrix metalloproteinase 9 protein were detected by 2,5-diphenyl-2H-tetrazolium bromide, Transwell and western blot. The dual-luciferase reporter gene experiment confirmed the relationship between microRNA-495-3p and fibroblast growth factor 9 targeting. Lidocaine could inhibit the proliferation, migration and invasion of human breast cancer cell line Michigan Cancer Foundation-7 cells, promote the expression of microRNA- 495-3p and inhibit the expression of fibroblast growth factor 9 protein in human breast cancer cell line Michigan Cancer Foundation-7 cells. Silencing microRNA-495-3p expression or fibroblast growth factor 9 overexpression decreased the inhibitory effect of lidocaine on proliferation, migration and invasion of human breast cancer cell line Michigan Cancer Foundation-7 cells. MicroRNA-495-3p negatively regulated fibroblast growth factor 9 expression in human breast cancer cell line Michigan Cancer Foundation-7 cells. Silencing fibroblast growth factor 9 expression reversed the effect of microRNA-495-3p silencing on proliferation, migration and invasion of human breast cancer cell line Michigan Cancer Foundation-7 cells treated with lidocaine. Lidocaine inhibits the expression of fibroblast growth factor 9 by up-regulating microRNA-495-3p, thus inhibiting the proliferation, migration and invasion of breast cancer cells.

Gas chromatography-mass spectrometry fingerprint and in vitro cytotoxic studies of Rubus steudneri leaf fractions against michigan cancer foundation-7 breast cancer cell line

Aim: Rubus steudneri Schweinf. (Rosaceae) is one of three Rubus species that grow in Ethiopia. Studies of this species have been restricted to in vitro antioxidant, antidiabetic, and nutritional evaluation. Until today, no identification has been made of its phytochemical fingerprints, resulting in an unclear picture of its phytochemical constituents. This study investigated the phytochemical composition of R. steudneri and its in vitro cytotoxicity against Michigan Cancer Foundation-7 (MCF-7) breast cancer cell lines and Vero cell lines. Materials and Methods: The leaf powder was subjected to liquid–liquid fractionation using n-hexane, chloroform, ethyl acetate, methanol, and water as solvents and the obtained fractions were subjected to flash chromatography and in vitro cytotoxicity studies in MCF-7 cell lines at concentrations from 1 to 1000 μg/mL, using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Results: The chloroform fraction proved to be the most promising of the bioactive fractions, with 50% of cytotoxicity inhibition for concentrations of 10 μg/mL on MCF-7 cell lines and 125 μg/mL on Vero cell lines. Two important drug leads, namely butyl isobutyl phthalate and 2-pyrrolidinone 5-(cyclohexylmethyl), were identified in chloroform fraction using gas chromatography-mass spectrometry (GC-MS) and may be responsible for the in vitro cytotoxicity. Flow cytometer results indicated that the chloroform fraction arrests cell cycle in the sub-G1 phase at a concentration of 100 μg/mL on MCF-7 cell lines, which proves that metabolites in this fraction may belong to the apoptotic population. Conclusion: In this study, butyl isobutyl phthalate and pyrrolidinone 5-(cyclohexylmethyl), which have significant cytotoxic effects and cell cycle arrest, are extracted. Further in-depth research is in progress to prove the anticancer activity of R. steudneri in search of new leads for anticancer drugs.

Effect of Sex Determining Region Y-Box 2 on the Biological Characteristics of Breast Cancer Cells and its Clinical Significance

To investigate the effects of the sex determining region Y-box 2 gene silencing on biological characteristics and the mitogen-activated protein kinase kinase kinase kinase 4 signal pathway of breast cancer cells. Firstly, the expression of sex determining region Y-box 2 in breast cancer was detected by the immunohistochemical method and the relationship with clinicopathological features was analyzed. Then the sex determining region Y-box 2 silencing breast cancer cell line Michigan Cancer Foundation-7 was prepared, and the messenger ribonucleic acid and protein expression levels of sex determining region Y-box 2, c-Jun N-terminal kinase, p53 and tumor necrosis factor-alpha in the mitogen-activated protein kinase kinase kinase kinase 4 pathway were detected by quantitative polymerase chain reaction and western blotting. Cell proliferation, migration, invasion and apoptosis were detected by the Cell Counting Kit-8, scratch test, transwell test, and flow cytometry. After si-sex determining region Y-box 2 transfection, the expression of sex determining region Y-box 2, c-Jun N-terminal kinase, p53 and tumor necrosis factor-alpha decreased, cell growth was inhibited, cell proliferation, migration and invasion decreased significantly, and apoptosis increased, while si-sex determining region Y-box 2+ mitogen-activated protein kinase kinase kinase kinase 4 transfection could reverse the above trend. Sex determining region Y-box 2 silencing can activate the mitogen-activated protein kinase kinase kinase kinase 4 signal pathway, promote the proliferation, migration, and invasion of breast cancer cells and inhibit apoptosis.

Porphyrin-based covalent organic framework as bioplatfrom for detection of vascular endothelial growth factor 165 through fluorescence resonance energy transfer

A fluorescent aptasensor based on porphyrin-based covalent organic framework (p-COF) and carbon dots (CDs) was constructed for detecting vascular endothelial growth factor 165 (VEGF165) and for imaging of the breast cancer cell line Michigan cancer foundation-7 (MCF-7). CDs synthesized with strong photoluminescence at λ∼380 nm were used as donors to label the VEGF165-targeted aptamers (AptVEGF/CDs). Additionally, the p-COF nanostructure comprised rich functional groups of CN on the surface and π-stacking planar nanostructure, resulting in the CDs adsorption via weakly π−π stacking, hydrogen bond and the Van der Waals force. Thereby, the fluorescence resonance energy transfer (FRET) occurred due to the close distance between the p-COF network and CDs, leading to the quenching of the fluorescence feature of CDs and p-COF. In the presence of VEGF165, the G-quadruplex was formed via the specific binding between VEGF165 and aptamer. It impelled that the release of partial VEGF165-AptVEGF/CDs complex, affording the fluorescence recovery of the sensing system to some extent. Consequently, the proposed AptVEGF/CDs/p-COF fluorescence biosensor offered excellent analytical performances for the VEGF165 detection, displaying a detection limit of 20.9 fg mL−1 within a wide linear range of the VEGF165 concentration of 1.0 pg mL−1–100 ng mL−1. The developed fluorescence biosensor was also used to determine VEGF165-overexpressed in MCF-7 cancer cells. Thereby, the present work can greatly widen the application of COFs in the development of aptasensors and cancer diagnosis.

Letrozole and zoledronic acid changed signalling pathways involved in the apoptosis of breast cancer cells

ObjectivesOestrogen plays a key role in the development of breast malignancies. Therefore, aromatase inhibitors (e.g. letrozole [LTZ]) are widely used in the treatment of breast cancer. On the other hand, oestrogen is important to the integrity of bone mass. Research has shown that zoledronic acid (ZLA) may prevent osteoporosis. Therefore, the present research aims to investigate the effect of a combination of LTZ and ZLA in the treatment of breast cancer and in reducing osteoporosis in patients with breast cancer. MethodsWe used immunocytochemistry and Western immunoblotting techniques in this study. ResultsWe observed that LTZ inhibited cellular growth of Michigan Cancer Foundation-7 (MCF-7) and T-47D at IC50 (70 ± 0.001) and (140 ± 0.004) nM, respectively, whereas ZLA inhibited cellular growth at IC50 (50 ± 0.005) μM and (150 ± 0.004) μM for MCF-7 and T-47D cell lines, respectively. Interestingly, the LTZ and ZLA combination down-regulated the protein expression of signal transducer and activator of transcription 3 (STAT3) and up-regulated BRCA1 protein expression in both cell lines. Moreover, a notable enhancement in the nuclear localisation of the BRCA1 protein was obtained after treatment of T-47D cells with LTZ for 24 h compared to the control cells. In contrast, there was a reduction in the nuclear localisation of STAT3 protein, which could be an attractive target for inhibition of breast cancer proliferation and progression. ConclusionOur study has shown that a combination of LTZ and ZLA enhanced apoptosis and inhibited growth of both breast cancer cell lines. This combination can be used to maintain bone integrity in women with breast cancer.

Electrochemically detecting DNA methylation in the EN1 gene promoter: implications for understanding ageing and disease.

There is a growing need for biomarkers which predict age-onset pathology. Although this is challenging, the methylome offers significant potential. Cancer is associated with the hypermethylation of many gene promoters, among which are developmental genes. Evolutionary theory suggests developmental genes arbitrate early-late life trade-offs, causing epimutations that increase disease vulnerability. Such genes could predict age-related disease. The aim of this work was to optimise an electrochemical procedure for the future investigation of a broad range of ageing-related pathologies. An electrochemical approach, which adopted three analytical techniques, was used to investigate DNA methylation in the engrailed-1 (EN1) gene promoter. Using synthetic single-stranded DNA, one technique was able to detect DNA at concentrations as low as 10 nM, with methylation status distinguishable at concentrations >25 nM. A negative correlation could be observed between % methylation of a heterogeneous solution and the key electrochemical parameter, charge transfer resistance (Rct; r = −0.982, P<0.01). The technique was applied to the breast cancer cell line Michigan Cancer Foundation-7 (MCF-7), where a similar correlation was observed (r = −0.965, P<0.01). These results suggest electrochemistry can effectively measure DNA methylation at low concentrations of DNA. This has implications for the future detection of age-related disease.

Nanocomposite of functional silver metal containing curcumin biomolecule model systems: Protein BSA bioavailability

Curcumin, a constituent of Curcuma longa L-Zingiberaceae is used in traditional Indian and worldwide medicine and shows anticancer and antioxidant properties. Curcumin has numerous biological and pharmacological activities but due to its hydrophobic nature, the major drawback is poor absorption and rapid elimination, rendering curcumin with the tag of a poor biomaterial. Hence, there is a need to develop functional metal containing curcumin model systems (FMCCMS) as a metallo-biomolecule to enhance the bioavailability of curcumin. We designed the interaction of silver metal ion with curcumin to form curcumin-silver nanocomposite (CURC-AgNCP) via ultrasonic synthetic route. Formations of FMCCMS were characterized by spectroscopic techniques. The crystalline face-centered cubic pattern and particle size of the nanocomposite was evaluated using X-ray diffraction and high-resolution transmission electron microscopy. The bonding of silver metal to curcumin was confirmed by X-ray photon spectroscopy. Interaction of the nanocomposite with bovine serum albumin (BSA) protein was performed using excitation, emission, and circular dichroism spectroscopy. In binding interaction of BSA, the negative value of ∆S° (−358.04 J mol−1 K−1) and ∆H° (−129.42 KJ mol−1) demonstrates the hydrophilic nature of the nanocomposite. The binding distance r evaluated according to the Forster resonance energy transfer theory and was 4.69 nm for CURC-AgNCP, which suggested non-radiative transfer of energy between CURC-AgNCP and BSA. The role of FMCCMS metallo-biomolecule CURC-AgNCP in medicine for cancer activity can have immense importance and hence we performed Sulphorhodamine B based in-vitro cytotoxicity assay on human breast cancer Michigan Cancer Foundation-7 cell line.

Mechanisms of crude protein from medicinal mushroom Ophiocordyceps sobolifera against human breast MCF-7 cancer cells

The mycelial extract of the entomopathogenic fungus Ophiocordyceps sobolifera has recently been reported to have anti-bacterial, anti-fungal and anti-cancer effects; however, there have been no reports on the medicinal value of the crude protein against the Michigan Cancer Foundation-7 (MCF-7) human breast cancer cell line. Therefore, in this study, crude proteins from 11 isolates of O. sobolifera were subjected to screening and evaluation for potential anticancer activity against a breast cancer cell line. The cytotoxic and antiproliferative effects of the crude proteins on human breast cancer MCF-7 cell line was investigated using the sulforhodamine B (SRB) assay and colony formation. The gene expression and cell cycle-associated protein levels were assayed by reverse transcription-quantitative polymerase chain reaction and western blotting analysis. The results showed that the crude proteins of all isolates had anti-cancer activity in a time- and dose-dependent manner. Among them, isolates Cod-KK1524, Cod-KK1630, Cod-KK1643 and Cod-KS1601 had greater inhibitory effects against cancer cells; therefore, these were chosen for further evaluation of anti-cancer mechanisms. These crude proteins exhibited a potent anticancer effect on the MCF-7 cells by inhibiting colony formation in a dose-dependent manner. Furthermore, the crude proteins significantly decreased the cyclin-dependent kinase 6 (cdk6) and cyclin D1 expression. Additionally, crude proteins significantly enhanced the potent cyclin-dependent kinase inhibitor (p21) levels, and reduced the expression level of the cell cycle protein, cyclin D1. These findings indicate that the crude protein of the entomopathogenic fungus O. sobolifera may be useful for developing an anti-cancer drug candidate for breast cancer therapeutic treatment.

Association of BRCA Mutations and BRCAness Status With Anticancer Drug Sensitivities in Triple-Negative Breast Cancer Cell Lines

BackgroundMany triple-negative breast cancers (TNBCs) show impaired breast cancer susceptibility gene I (BRCA1) function, called BRCAness. BRCAness tumors may show similar sensitivities to anticancer drugs as tumors with BRCA1 mutations. In this study, we investigated the association of BRCA mutations or BRCAness with drug sensitivities in TNBC. MethodsBRCAness was evaluated as BRCA1-like scores, using multiplex ligation-dependent probe amplification in 12 TNBC cell lines, including four with mutations. Sensitivities to docetaxel, cisplatin, and epirubicin were compared with BRCA mutations and BRCA1-like scores. Cisplatin sensitivity was examined in BRCA1 knockdown Michigan Cancer Foundation-7 cell lines. ResultsEight and four cell lines had characteristics of BRCAness and non-BRCAness, respectively. The 50% inhibitory concentration of docetaxel was higher in BRCA mutant and BRCAness cell lines than their counterparts. BRCA1-like scores showed a weak positive correlation with docetaxel sensitivity (r = 0.377; P = 0.039). Regarding cisplatin, scores were lower in BRCA mutants and BRCAness tumors than their counterparts. A negative correlation was found between BRCA1-like scores and cisplatin sensitivity (r = −0.407; P = 0.013). No differences were found for epirubicin. BRCA1 gene knockdown increased the cisplatin sensitivity of Michigan Cancer Foundation-7 cells. ConclusionsBRCA1-like scores were associated with cisplatin sensitivity and docetaxel resistance. BRCA1-like score is hence a promising indicator for estimating drug sensitivities in TNBC.

Cell phone and wireless radiation hazard on TRPV1 channel activation: A Scoping Review study

TRPV1 channel is activated with pH changes, increasing temperature, free radicals, and oxidative stress, and it may be a good measure for detecting the effect of microwave non-ionizing radiation on cellular and molecular responses. The scoping review study aims to survey the influence of mobile and wireless radiation on the TRPV1 channel mechanism. We systematically considered international databases to recognize related studies, including PubMed, Scopus, Cochrane Library, CINAHL, ISI Web of Science, Science Direct from the start, PROSPERO and EMBASE use headings for scientific topics (Mesh). The hunt covered the period from 1980 until 2019. A total of seven studies comprising 137 cases, three in vitro tests on breast cancer cell line Michigan Cancer Foundation-7 (MCF-7), embryonic kidney HEK293T cells and Rabbit polyclonal anti-TRPV1, and four in vivo experimental studies comprising 21 male and 24 female Wistar Albino rats, were included. Two studies indicated cell phones induced mitochondrial ROS, Oxidative stress-induced in the hippocampus, mitochondrial apoptosis via activation of TRPV1 in HIPPON and DRGN cells. The cytosolic Ca2+ induced by TRPV1 induced oxidative overactions and apoptosis at a distance of 10 cm from the source of the cells. Our review shows that cell phone and wireless radiation have a possible effect on Ca2+ signaling by TRPV1 channel activation. This reaction may induce ROS, Fenton and Haber-Weiss reaction which increase free radicals’ level in the mitochondria.

Rasam (South Indian Spice Soup) - Attenuates the Mammary Tumor Induction Magnitude of 7,12-Dimethylbenz[a] anthracene in Sprague–Dawley Rats

Background: Recently, we have reported rasam (South Indian spice soup) for antiproliferative activity against Michigan Cancer Foundation-7 cell lines. Objectives: Breast cancer seems to be more common in the younger age group. Considering the deviation of the younger group from the traditional food habits as one of the many reasons of breast cancer incidence, rasam was investigated for its chemopreventive effect on mammary carcinoma. Materials and Methods: Rasam at 3 and 4 mL/kg dose was administered to female Spraque–Dawley rats for 30 days before 7,12-dimethylbenz[a] anthracene (DMBA) induction of mammary carcinoma. Tamoxifen was used as the standard drug. Body weight, thiobarbituric acid reactive substances (TBARSs), phase I enzymes, hexokinase (HK), aldolase (AD), glucose 6-phosphatase (G6P), fructose 1,6-bisphosphatase (F16P), citric acid cycle (CAC) enzymes, histopathology, tumor weight, tumor latency, and tumor incidence were studied. Results: In all the studied parameters such as body weight, TBARS, phase I enzymes, HK, AD, G6P, F16P, and CAC enzymes, rasam-pretreated groups showed better prevention than tamoxifen-treated groups. Histochemical findings also clearly confirm the chemopreventive effect of rasam. Particularly at the dose of 4 mL/kg, rasam was efficient in reducing the percentage of incidence of tumor, number of tumor, and tumor weight. Conclusion: The present study reveals that rasam attenuates the mammary tumor induction magnitude of DMBA in female rats.

English

Maerua pseudopetalosa (Gilg and Bened.) De Wolf tubers which are used traditionally as antitumor agent in Sudan were subjected to separation by column chromatography technique. Eight fractions were obtained for the ethyl acetate extract and twelve for the ethanolic extract. The ethanolic fractions F8, F9, F11 and F12, with high bioactivity were subjected to further investigations. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was used for assessment of the cytotoxicity. Remarkable cytotoxicity against Michigan Cancer Foundation-7 (MCF-7) was shown, for the first time. Actually, the results revealed that F12 is a very promising one with remarkable activity against MCF-7 cell lines (43.51 µg/g at 72 h), high antioxidant activity (91.3% by 1,1-diphenyl-2-picryl-hydrazyl (DPPH) and 97% by ABTS) and flavonoid and phenolic contents (11.75 and 20.72 mg/g). Six compounds were detected in F9 (syrngic 11.65 µg/g, sinapic 8 µg/g) and F12 (gallic 88.12 µg/g, caffic 11.1 µg/g, sinapic 38.67 µg/g) which were not recorded in any previous work in the available literature by using high performance liquid chromatography (HPLC). Key words: Michigan Cancer Foundation-7 (MCF-7) breast cancer, Maerua pseudopetalosa, phenol and flavonoid contents, high performance liquid chromatography (HPLC) analysis, antioxidant activity.

Green synthesis, characterization and antibacterial activity of silver nanoparticles by Malus domestica and its cytotoxic effect on (MCF-7) cell line

This article reports the utilization of Malus domestica for the synthesis of silver nanoparticles (AgNPs) with cytotoxic activity against the Michigan Cancer Foundation-7 (MCF-7) cell line as well as their antibacterial and radical scavenging potential. The biosynthesized AgNPs were confirmed using various analytical characterization techniques. The cytotoxic effect of Malus domestica-AgNPs (M.d-AgNPs) was studied by MTT assay and scavenging efficacy was assessed by DPPH, nitric oxide radical and phosphomolybdate assays. Furthermore, green synthesized nanoparticles were evaluated for their antibacterial activity against multidrug resistant-clinical isolates. M.d-AgNPs were observed to be almost spherical in shape with an average diameter from 50 to 107.3 nm as assessed by TEM and DLS. M.d-AgNPs revealed the dose-dependent antioxidant activity and antimicrobial activity against multidrug-resistant bacterial strain viz. Enterobacter aerogenes, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Escherichia coli. Also, in vitro studies revealed dose-dependent cytotoxic effects of M.d-AgNPs treated MCF-7 cell line. The data strongly suggest that M.d-AgNPs had a potential antioxidant, antimicrobial and cytotoxicity activity.

Avoiding the Interference of Doxorubicin with MTT Measurements on the MCF-7 Breast Cancer Cell Line.

Doxorubicin (DOXO) is an adjuvant chemotherapy agent and is also commonly used in cell biology research. Cytotoxic assays in cell culture are frequently used in order to stablish drug concentrations that are useful for controlling cell proliferation. One common cytotoxic method used is 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT). Our present research aims to support future studies in engaging MTT assay using DOXO that exhibits a strong red coloration and fluorescence, and so it is assumed that DOXO may interfere with commonly used colorimetric assays such as MTT. The interference of DOXO in the MTT determination was evaluated in a Breast Cancer cell line Michigan Cancer Foundation-7 (MCF-7). The interference was evaluated by means of spectroscopic methods in particular spectrophometry and fluorescence spectroscopy of MTT and DOXO. We postulate that the medium and the MTT reagent itself can interfere on the metabolic activity method, so in order to achieve better results, DMEM was replaced by a neutral buffer like Phosphate-buffered saline (PBS). This protocol may be extremely useful in future studies involving DOXO.

DESIGN, SYNTHESIS, MOLECULAR DOCKING, AND BIOLOGICAL EVALUATION OF PYRAZOLE 1-CARBOTHIAMIDE INCORPORATED ISOXAZOLE DERIVATIVES

Objectives: Novel isoxazole incorporated pyrazole carbothiamide 5 (a-r) was designed and synthesized, docked and evaluated for anticancer activity Michigan Cancer Foundation-7 (MCF-7), and breast cancer cell lines.&#x0D; Materials and Methods: Designed compounds were synthesized by the condensation of 1-(5-methyl-3-(4-nitrophenyl) isoxazole-4-yl) -3-(substitutedphenyl) prop-2-en-1-one (4) with thiosemicarbazides and substituted thiosemicarbazides to give the target molecules 5 (a-r). To predict the affinity and activity of the ligand molecule, the docking program Accelrys Discovery Studio 2.1 was employed to generate different bioactive binding poses of designing molecules at the active site of human Dihydrofolate Reductase (DHFR) (PDB ID: 1KMS). All the synthesized compounds were characterized based on the spectral and elemental analysis data. Antiproliferative activity was performed against MCF-7 breast cancer cell lines.&#x0D; Results: All the synthesized compounds showed the characteristic peaks in Fourier-transform infrared,1H C13NMR, and mass spectral analysis. During docking, all the synthesized compounds 5 (a-r) exhibited higher fitness scores with minimum three bonding interaction with the active site human DHFR (PDB ID: 1KMS). In the MTT assay based on MCF-7 breast cancer cell lines, most of the compounds exhibited significant activity. In the antiproliferative assay against MCF-7 cell lines, most of the compounds exhibited potent activity with IC50 values in micromolar concentrations. Compounds 5a, 5b, 5f, 5h, and 5k have exhibited significant anticancer activity.&#x0D; Conclusions: The derivatives were synthesized in quantitative yields. New derivatives possess the antiproliferative activity and antitubercular activity.

In vitro cytotoxic activity of Zaleya Pentandra L. Extracts against the breast cancer adenocarcinoma cell line MCF-7.

To evaluate the cytotoxicity of crude aqueous and ethanol extracts of Zaleya pentandra against oestrogen receptor-positive breast cancer cell line Michigan Cancer Foundation-7. The study was conducted at the Institute of Chinese Medicine, the Chinese University of Hong Kong, Hong Kong, from March to September 2017, and comprised Zaleya pentandra herbaceous perennial plant collected from Pakistani cities of Shakargarh, Lahore and Sialkot. Both aqueous and ethanol extracts were prepared in solvents following Soxhlet extraction technique. Rate of reduction in viability of cancer cell line was studied through MTT (dye compound 3-(4,5-Dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide) assay on the basis of time of incubation and the dose of extract. Analysis was performed using High Performance Liquid Chromatography with Diode Array Detector in order to found the compounds responsible for the death of cancer cells. Cell viability was observed to be dose-dependent (range: 16.7-37.4%) for aqueous extracts. Minimum inhibition concentration was 16.65% at 200μg/ml after 24 hours of incubation, whereas maximum inhibition was 37.39% at 3200μg/ml. Ethanol extracts showed less inhibition, with maximum inhibition being 25.29% at 1000μg/ml and minimum 13.57% at 62.5μg/ml. Certain polar compounds, like Hydroxytrosol and Tyrosol, could be obtained from the aqueous extracts only. Zaleya pentandra aqueous extract was found to have potential benefit towards cytotoxicity of breast cancer cells.

MiRNA-192-5p impacts the sensitivity of breast cancer cells to doxorubicin via targeting peptidylprolyl isomerase A.

The administration of doxorubicin (DOX) is one of the first-line treatments of breast cancer. However, acquisition of resistance remains the major obstacle restricting the clinical application of DOX. MicroRNAs (miRNAs) are small, noncoding RNAs which play crucial role in epigenetic regulation. Recent studies have shown that miRNAs are associated with tumor chemoresistance. Here we aim to explore the role of miRNA-192-5p in resistance to DOX in breast cancer cells. Normal human breast epithelial cell line MCF-10A, breast cancer cell line Michigan Cancer Foundation-7 (MCF-7), and DOX-resistant breast cancer cell line MCF-7/ADR were used here. The expression of miR-192-5p was examined by qPCR, and the expression of peptidylprolyl isomerase A (PPIA) was examined by qPCR and Western blot. The effects of miR-192-5p overexpression on the sensitivity to DOX were confirmed by Methylthiazolyldiphenyl-tetrazolium bromide (MTT) and Annexin-V/PI assay. Downstream molecular mechanisms, including PPIA, BAD, CASP9, Bcl-2, and c-Jun N-terminal kinase (JNK) activation, were detected by Western blot and qPCR. Luciferase reporter assay was used to validate the association between miR-192-5p and PPIA. miR-192-5p was downregulated while PPIA was upregulated in MCF-7/ADR cells. Functionally, miR-192-5p overexpression increased sensitivity to DOX by promoting cell apoptosis. Mechanistically, miR-192-5p overexpression performed its function by activating JNK, augmenting BAD and caspase9 expression, and suppressing Bcl-2 and PPIA expression. Luciferase assay validated that PPIA was a direct target of miR-192-5p. miR-192-5p sensitizes breast cancer cells to DOX by targeting PPIA, suggesting that miR-192-5p might serve as a novel target for reversing DOX resistance and controlling breast tumor growth.