Activity In MCF-7 Cells Research Articles

Water-soluble propolis and bee pollen of Trigona spp. from South Sulawesi Indonesia induce apoptosis in the human breast cancer MCF-7 cell line.

Bee products are best known as one of the beneficial natural products providing multiple pharmacological effects, such as antimicrobial, antiviral, anti-inflammatory and anticancer effects. The present study aimed to identify potent products derived from the stingless bee Trigona spp. from Luwu Utara (South Sulawesi, Indonesia), focussing on the water-soluble extract of propolis and bee pollen, against the proliferation of the human breast cancer MCF-7 cell line. The results from DPPH (2,2-diphenyl-1-picrylhydrazyl) method of antioxidant assay revealed that water-soluble propolis and bee pollen had high antioxidant activity, with half-maximal effective concentrations against DPPH radicals of 1.3 and 0.4 mg/ml, respectively. Additionally, water-soluble propolis and bee pollen exhibited a significant antiproliferative activity in MCF-7 cells, with IC50 values of 10.8±0.06 and 18.6±0.03 mg/ml, respectively (P<0.05). Significant cytotoxic effects were observed after 24 h of treatment via microscopic and flow cytometric analysis, where a morphological change toward late apoptosis was observed. By contrast, honey had low antioxidant activity and no antiproliferative effect in MCF-7 cells. The water-soluble propolis also exerted its antiproliferative effect in the human keratinocyte HaCaT cell line. The antiproliferative activity was similar (P>0.05) at 24 and 48 h of treatment, with IC50 at 2.7±0.06 mg/ml and <0.4 mg/ml, respectively. Notably, bee pollen was less toxic to HaCaT cells after 24 h of treatment than the water-soluble propolis, with IC50>50 mg/ml. Its antiproliferative activity was significantly increased after 48 h of treatment, with IC50 at 9.6±0.07 mg/ml (P<0.05). In addition, similar to other poplar propolis, the high-performance liquid chromatography-ultraviolet and electrospray ionisation mass spectrometry analyses revealed that caffeic acid phenethyl ester was not the main bioactive compound of the samples examined. Furthermore, two major proteins (between ~50 and 75 kDa) were identified in the water-soluble propolis and bee pollen. The present results suggested that water-soluble propolis and bee pollen may have the potential to be elaborated further as a breast anticancer therapy.

Construction of a Universal and Label-Free Chemiluminescent Sensor for Accurate Quantification of Both Bacteria and Human Methyltransferases.

DNA methylation plays important roles in various biological processes, and the alteration of DNA methyltransferase activity can induce the aberrant DNA methylation patterns. Despite the progress in methyltransferase activity assays, few methods enable the detection of both bacteria and human methyltransferases. Herein, we construct a universal and label-free chemiluminescent sensor for accurate quantification of both bacteria methyltransferases (e.g., M. SssI methyltransferase (M.SssI MTase)) and human methyltransferases (e.g., DNA (cytosine-5)-methyltransferase 1, (Dnmt1)) by integrating a dumbbell probe with BssHII endonuclease-mediated rolling circle amplification (RCA). We ingeniously design a structure-switchable dumbbell probe which integrates target-recognition, BssHII endonuclease-cleavage, RCA amplification and signal transduction in one probe for the detection of both M.SssI MTase and Dnmt1. Moreover, the introduction of two BssHII endonuclease recognition sites in a dumbbell probe can greatly reduce the false positivity resulting from the incomplete cleavage of dumbbell probe by BssHII, because once one of two recognition sites is identified by BssHII, the dumbbell probe can be completely digested by Exonuclease III (Exo III) and Exonuclease I (Exo I) to prevent the nonspecific RCA. This chemiluminescent sensor can accurately quantify M.SssI MTase in both 10% serum and various cell lysis buffers, and even sensitively detect Dnmt1 activity in MCF-7 cells. Furthermore, this chemiluminescent sensor can be used to screen the inhibitors of Dnmt1 and M.SssI MTase, with promising applications in disease diagnosis and drug discovery.

Lanthanide(III) complexes with N-(2,6-dimethylphenyl)oxamate and 1,10-phenanthroline: Synthesis, characterisation and cytotoxicity against MCF-7, HEC-1A and THP-1 cell lines

Seven novel mixed-ligand complexes, [Ln(Hpma)2(phen)2(OAc)]·2H2O (Ln = La (1), Ce (2) and Nd (3)), [Er(Hpma)2(phen)(OAc)(H2O)]·EtOH·H2O (4), [Yb(Hpma)3(phen)]·2EtOH (5) and [Ln(Hpma)3(phen)(H2O)]·4H2O (Ln = Nd (6) and Gd (7)) (Hpma = N-(2,6-dimethylphenyl)oxamate; phen = 1,10-phenanthroline) were prepared in the presence of NaOH. Complexes 1–3 are isostructural and each metal ion is ten-coordinate, adopting a distorted sphenocorona geometry, with continuous shape measures of 2.702, 2.677 and 2.592, respectively. In contrast, the nona-coordinate complexes 4, 6 and 7 displayed spherical capped square antiprism molecular geometry, while the Yb(III) complex is bonded to three bidentate Hpma– ligands and one phen, forming a square antiprism structure (coordination number = 8). Besides the strong Ln(III)-ligand bonds as the main driving force for coordination, the relatively weak hydrogen bonds (OH∙∙∙O and NH∙∙∙O), CH∙∙∙O contacts, CH∙∙∙π, NH∙∙∙π and offset π-π interactions between the aromatic rings, play a key role in crystal packing. The UV–Vis-NIR absorption spectra of the metal complexes are associated with the π → π* transitions of phen and the Laporte-forbidden f-f transitions of the Nd(III), Er(III) and Gd(III) ions from the 4I9/2, 4I15/2 and 8S7/2 ground states, respectively, to the various excited J-levels. Cytotoxicity studies revealed antiproliferative activity in MCF-7 cells (p < 0.05) after treatment with phen and coordination compounds 1–7. Statistically significant cytotoxic effects were also exerted by Nd(NO3)3·6H2O, phen, 3, 6 and 7 in HEC-1A cells (p < 0.05). No cytotoxic effects were observed in THP-1 cells (p > 0.05) for all the compounds, except for the positive control curcumin; indicating that the target compounds do not exert adverse effects on the immune system monocytes.

A natural flavonoid, apigenin isolated from Clerodendrum viscosum leaves, induces G2/M phase cell cycle arrest and apoptosis in MCF-7 cells through the regulation of p53 and caspase-cascade pathway.

With 9.6 million deaths in 2018, cancer remains the second leading cause of death worldwide. Breast cancer is the most deadly type of cancer among females, with 55.2% of crude incidence rate and 16.6% of crude mortality rate. The present study was aimed to investigate the anti-breast cancer potential of natural dietary flavonoid, apigenin isolated from Clerodendrum viscosum leaves. Apigenin was evaluated for in-depth anticancer activity in MCF-7 cells using cell viability assay, cell cycle analysis, Annexin-V-FLUOS staining, ROS induction, morphological analysis, and western blot analysis. Apigenin showed selective cytotoxicity on MCF-7 cells with an IC50-56.72 ± 2.35µM, while negligible cytotoxicity was observed on WI-38 cells. Further, the flow cytometer-based analysis showed that apigenin halted MCF-7 cells in the G2/M phase arrest followed by dose-dependent apoptosis. Moreover, the FACS and confocal microscopy results confirmed the elevation of intracellular ROS and nuclear fragmentation in apigenin-treated MCF-7 cells. Western blots showed up-regulation of cell cycle regulatory proteins, increased p53 expression, Bax/Bcl-2 ratio, activation of caspases, and cleavage of PARP. Finally, apigenin treatment in the presence of Pifithrin-µ showed decreased apoptotic population and it was further confirmed through western blotting study. The results revealed the vital role of p53 in apigenin-induced apoptosis in MCF-7 cells. In the present findings, treatment of apigenin-induced intracellular ROS in MCF-7 cells followed by induction of G2/M phase cell cycle arrest and further apoptosis through the regulation of p53 and caspase-cascade signaling pathway.

The Synthesis, Anticancer Activity, Structure-Activity Relationships and Molecular Modelling Studies of Novel Isoindole-1,3(2H)-dione Compounds Containing Different Functional Groups.

Isoindole-1,3(2H)-dione derivatives are known to have cytotoxic effects on many cancer cells. The anticancer activity of these compounds varies depending on the substituents attached to them. Therefore, the effect of substituents is very important when determining the anticancer activities of molecules. We have recently reported an example of the substituent effect. According to that work, the anticancer activity against HeLa, C6, and A549 cancer cell lines of isoindole- 1,3(2H)-dione compounds containing tert-butyldiphenylsilyl ether, azido, and hydroxyl groups was examined by our group. It was found that an isoindole-1,3(2H)-dione compound containing both tert-butyldiphenylsilyl ether group and azido groups showed higher anticancer activity than 5-fluorouracil and another isoindole-1,3(2H)- dione compound containing both azido and hydroxyl groups. After we discovered that tert-butyldiphenylsilyl ether group in the skeletal structure of isoindole-1,3(2H)-dione exhibits anticancer activity against HeLa, C6, and A549 cancer cell lines, we wanted to examine the anticancer activities of different silyl ether groups, i.e., OTMS, -OTBDPS, and -OTBDMS groups, and also -OH and -Br groups, by comparing them with each other according to the structure-activity relationship. All of the synthesized compounds were characterized by 1H and 13C NMR spectra, IR spectroscopy, and mass spectra measurements. The IC50 values of these compounds were calculated for all cancer cell lines and compared with each other and cisplatin, which is a platinum-containing chemotherapeutic drug. Molecular modelling studies were carried out using the MOE software package. It was found that compounds 13 and 16, containing both silyl ether (-OTBDMS) and -Br groups, showed higher anticancer activity than cisplatin against both Caco-2 and MCF-7 cell lines. Compounds 20 and 23 showed anticancer activity in MCF-7 cells and compounds 8, 9, 20, and 23 in Caco-2 cells. While compounds 20 and 23 have only a silyl ether (-OTMS) group, compounds 8 and 9 have only a -OH group. Molecular modelling studies indicated that compounds 8 and 13, as well as their analogs, may bind to the active site of hRS6KB1 (pdb: 4l3j), compound 11 may bind to the active site of human mTOR (pdb: 4jt5) and additionally, compounds 10-17 are expected to be both mutagenic and reactive according to the mutagenicity and reactivity calculations. According to these results, the anticancer activities of isoindole-1,3(2H)-dione compounds (8 - 23) vary depending on the groups they contain and these groups affect each other's activities. Silyl ethers (-OTBDMS and -OTMS) and -OH and -Br groups in the skeletal structure of isoindole-1,3(2H)-dione can be regarded as anticancer agents. In this sense, compounds 13 and 16, containing both silyl ether (-OTBDMS) and - Br groups, may be regarded as alternative chemotherapeutic drugs. This work may lead to the synthesis of new isoindole-1,3(2H)-dione compounds containing different silyl ether groups and studies evaluating their anticancer activities or other biological properties.

Abstract P4-08-04: Downregulation of DYRK2 contributes to tumor cell proliferation by enhancing CDK14 expression in breast cancer

Abstract Introduction: Tumor progression is the main cause of death in patients with breast cancer. Accumulating evidence suggests that dual-specificity tyrosine-regulated kinase 2 (DYRK2) functions as a tumor suppressor by regulating cell survival, differentiation, proliferation and apoptosis. However, little is known about the mechanisms of transcriptional regulation by DYRK2 in cancer progression, particularly with respect to cancer proliferation and invasion. Here, using a comprehensive expression profiling approach, we show that cyclin-dependent kinase 14 (CDK14) is a target of DYRK2. The aim of this study is to clarify whether DYRK2 suppresses the proliferation of breast cancer cells through CDK14 expression. Methods: Cell lines: We established stable DYRK2-depleted cells. MCF-7 cells were transfected with pSuper-puro vector (pSuper control) or pSuper-puro DYRK2 shRNAs (shDYRK2) along with puromycin to isolate stable cell lines. We generated stable DYRK2-overexpressing MDA-MB-231 cells using GFP-DYRK2 (GFP-DYRK2 cells). A GFP vector was transfected as a vehicle control (GFP control cells) with G418 to isolate stable cell lines. Moreover, we established both stable DYRK2- and CDK14-depleted cells. The shDYRK2 cells were transfected with the pSuper-neo vector (shDYRK2-pSuper control) or pSuper-neo CDK14 shRNAs (shDYRK2-shCDK14). The pSuper control cells were transfected with the pSuper-neo vector (pSuper control-pSuper control cells) as a vehicle control with puromycin and G418 to isolate stable cell lines. These manipulated cells were compared with the control cells by various assays. Immunohistological staining: Sixty samples from surgically treated breast cancer patients were obtained from the Surgery Department at the Jikei University Hospital between 2001 and 2002. The Jikei University School of Medicine Ethics Review Committee approved the study protocol, and informed consent was obtained. Results: We analyzed cancer cell proliferation by MTS and colony formation assays. We investigated the effects of CDK14 in DYRK2-depleted cells in a breast tumor xenograft model. We found that reduced DYRK2 expression increased CDK14 expression to promote cancer cell proliferation in vitro and tumorigenicity in vivo. We further identified androgen receptor (AR) as a candidate of DYRK2-dependent transcription factors to control CDK14. Real-time RT-PCR and western blotting revealed that inhibition of AR activity using an AR inhibitor MDV3100 decreased both mRNA and protein levels of CDK14 in shDYRK2 and GFP control cells. In contrast, knocking down DYRK2 induced AR transcription activity in MCF-7 cells. shDYRK2 cells were more sensitive to MDV3100 compared with pSuper control cells. Immunohistological staining of sixty samples from surgically treated breast cancer patients showed an inverse correlation between DYRK2 and CDK14 expression. Conclusion: These findings demonstrate that reduced DYRK2 expression in breast cancer promotes tumor cell proliferation by modulating CDK14 expression via AR. Citation Format: Yoshimi Imawari, Rei Mimoto, Noriko Yamaguchi, Makiko Kamio, Hiroko Nogi, Ken Uchida, Hiroshi Takeyama, Kiyotsugu Yoshida. Downregulation of DYRK2 contributes to tumor cell proliferation by enhancing CDK14 expression in breast cancer [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P4-08-04.

Design, synthesis, and molecular docking of novel 2-arylbenzothiazole multiangiokinase inhibitors targeting breast cancer.

A novel series of 2-arylbenzothiazoles 9, 10, and 12 were designed and synthesized as VEGFR-2/FGFR-1/PDGFR-β multiangiokinase inhibitors targeting breast cancer. Structural elongation of the known 2-phenylbenzothiazole scaffold (type I protein kinase inhibitor [PKI]), was carried out to afford series of type II PKIs 9, 10, and 12. Compounds 9d, 9f, 9i, and 9k exhibited potent multikinase inhibitory activity with IC50 values of 0.19, 0.18, 0.17, and 0.13 μM, respectively, against VEGFR-2; IC50 values of 0.28, 0.37, 0.19, and 0.27 μM, respectively, against FGFR-1; and IC50 values of 0.07, 0.04, 0.08, and 0.14 μM, respectively, against PDGFR-β. Moreover, the synthesized benzothiazoles demonstrated promising cytotoxic activity against the MCF-7 cell line. The most potent benzothiazoles 9d and 9i exhibited IC50 values of 7.83 and 6.58 μM, respectively, on the MCF-7 cell line in comparison to sorafenib (III), which showed IC50 = 4.33 μM. Additionally, 9d and 9i showed VEGFR-2 inhibitory activity in MCF-7 cells of 81% and 83% when compared with sorafenib (III), which showed 88% inhibition. Molecular docking of the designed compounds in the VEGFR-2 and FGFR-1 active sites showed the accommodation of the 2-phenylbenzothiazole moiety, as reported, in the hinge region of the receptor tyrosine kinase (RTK)-binding site, while the amide moiety is involved in hydrogen bond interactions with the key amino acids in the gate area; this in turn directs the aryl group to the hydrophobic allosteric back pocket of the RTKs in a type II-like binding mode. The synthesized benzothiazoles showed satisfactory ADME properties for further optimization in drug discovery.

Zn(ii), Cd(ii) and Hg(ii) saccharinate complexes with 2,6-bis(2-benzimidazolyl)pyridine as promising anticancer agents in breast and lung cancer cell lines via ROS-induced apoptosis.

New Zn(ii), Cd(ii) and Hg(ii) complexes of saccharinate (sac) and 2,6-bis(2-benzimidazolyl)pyridine (bzimpy), [Zn(bzimpy)2](sac)2·2H2O (Zn), [Cd(sac)2(bzimpy)] (Cd) and [Hg(sac)2(bzimpy)] (Hg), were prepared and fully characterized by spectroscopic methods and X-ray crystallography. In vitro anticancer screening in A549 (lung), MCF-7 (breast) and HT29 (colon) cell lines showed that Zn was highly cytotoxic against A549 and MCF-7 cells with IC50 values of 1.74 ± 0.06 and 3.15 ± 0.10 μM, respectively, and Hg demonstrated potent cytotoxic activity in MCF-7 cells (8.61 ± 0.98 μM), while Cd and bzimpy exhibited moderate growth inhibitory activities in all of the cell lines. In addition, they showed significantly lower toxicity towards normal human breast epithelial MCF10A cells. Moreover, the complexes exhibited significantly high nuclease activity towards plasmid DNA and their interactions with DNA were assessed by gel electrophoresis and DNA docking. Zn and Hg induced G0/G1 cell arrest and apoptotic cell death detected via typical DNA condensation/fragmentation, annexin V staining and caspase 3/7 activity in A549 and MCF-7 cells. These complexes further caused depolarization of mitochondria and oxidative damage of genomic DNA following excessive production of reactive oxygen species (ROS).

Rational design of ERα targeting hypoxia turn-on fluorescent probes with antiproliferative activity for breast cancer.

The overexpression of estrogen receptor (ER) α is not only closely related to the development of ER+ breast cancer, but is also an important biomarker for clinical diagnosis and treatment. Herein, we report several ERα targeting hypoxia turn-on fluorescent probes with antitumor activity for breast cancer cells. Among them, probes 3 and 5 displayed good ERα targeting ability and favorable hypoxia turn-on response in MCF-7 cells. Moreover, the probes 3 and 5 exhibited good antiproliferative activity towards MCF-7 cells (IC50 = 8.5 μM, 10.3 μM) and a much lower cytotoxicity to normal cells compared with the positive control. It is expected that these novel fluorescent probes may provide useful tools for the theranostics of ER+ breast cancer.

&lt;p&gt;Brevilin A Inhibits STAT3 Signaling and Induces ROS-Dependent Apoptosis, Mitochondrial Stress and Endoplasmic Reticulum Stress in MCF-7 Breast Cancer Cells&lt;/p&gt;

PurposeBreast cancer is the most common malignancy among women across the globe. Despite concerted efforts to improve the prevailing treatment modalities, the overall prognosis of breast cancer remains unsatisfactory. Recently, antiproliferative activity of Brevilin A (Brv-A), a sesquiterpene lactone compound of Centipeda minima, has been unveiled in various cancer types. Here, we have explored anticancer activity of Brv-A in MCF-7 breast carcinoma cells by targeting various pathways.Materials and MethodsCell proliferation rate was determined by CCK-8 and clonogenic assay. Cellular morphological changes were observed under phase contrast microscope while calcein-AM and PI was used for live/dead assay. Cell cycle assay was performed by flow cytometry. Apoptotic cell percentage was determined by Hoechst 33258 staining and flow cytometric analysis. ROS generation and mitochondrial membrane potential were measured using commercially available kits while protein expression was measured by Western blotting.ResultsIn our study, Brv-A exerted antiproliferative effect through mitotic arrest at G2/M phase of cell cycle and induced apoptosis in MCF-7 cells in a dose-dependent manner. Induction of apoptosis by Brv-A was found to be associated with ROS generation by targeting NOX2 and NOX3, mitochondrial dysfunction (MMP dissipation and Bcl-2 family proteins modulation), DNA fragmentation, JNK and p38 MAPK activation, endoplasmic reticulum (ER) stress by increasing Bip/GRP78, ATF4 and CHOP protein expressions and inhibition of STAT3 activation via decreased phosphorylation of JAK2 and SRC. Pretreatment of NAC, a ROS scavenger, partially reversed the aforesaid cellular events indicating ROS generation as the primary event to modulate cellular targets for induction of apoptosis. Besides, Brv-A has also been documented for inhibition of cell migration via decrease in COX-2 and MMP-2 expression.ConclusionTaken together, Brv-A induces G2/M phase arrest, ROS-dependent apoptosis, ER stress, mitochondrial dysfunction and inhibits STAT3 activation in MCF-7 cells signifying it to be one of the potential anticancer therapeutics in future.

3D-microenvironments initiate TCF4 expression rescuing nuclear β-catenin activity in MCF-7 breast cancer cells

Mechanical cues sensed by tumor cells in their microenvironment can influence important mechanisms including adhesion, invasion and proliferation. However, a common mechanosensitive protein and/or pathway can be regulated in different ways among diverse types of tumors. Of particular interest are human breast epithelial cancers, which markedly exhibit a heterogeneous pattern of nuclear β-catenin localization, a protein known to be involved in both mechanotransduction and tumorigenesis. β-catenin can be aberrantly accumulated in the nucleus wherein it binds to and activates lymphoid enhancer factor/T cell factor (LEF/TCF) transcription factors. At present, little is known about how mechanical cues are integrated into breast cancer cells harboring impaired mechanisms of β-catenin's nuclear uptake and/or retention. This prompted us to investigate the influence of mechanical cues on MCF-7 human breast cancer cells which are known to fail in relocating β-catenin into the nucleus due to very low baseline levels of LEF/TCFs. Exploiting three-dimensional (3D) microscaffolds realized by two-photon lithography, we show that surrounding MCF-7 cells have not only a nuclear pool of β-catenin, but also rescue from their defective expression of TCF4 and boost invasiveness. Together with heightened amounts of vimentin, a β-catenin/TCF-target gene regulator of proliferation and invasiveness, such 3D-elicited changes indicate an epithelial-to-mesenchymal phenotypic switch of MCF-7 cells. This is also consistent with an increased in situ MCF-7 cell proliferation that can be abrogated by blocking β-catenin/TCF-transcription activity. Collectively, these data suggest that 3D microenvironments are per se sufficient to prime a TCF4-dependent rescuing of β-catenin nuclear activity in MCF-7 cells. The employed methodology could, therefore, provide a mechanism-based rationale to dissect further aspects of mechanotranscription in breast cancerogenesis, somewhat independent of β-catenin's nuclear accumulation. More importantly, by considering the heterogeneity of β-catenin signaling pathway in breast cancer patients, these data may open alternative avenues for personalized disease management and prevention. Statement of significanceMechanical cues play a critical role in cancer pathogenesis. Little is known about their influence in breast cancer cells harboring impaired mechanisms of β-catenin's nuclear uptake and/or retention, involved in both mechanotransduction and tumorigenesis. We engineered 3D scaffold, by two-photon lithography, to study the influence of mechanical cues on MCF-7 cells which are known to fail in relocating β-catenin into the nucleus. We found that 3D microenvironments are per se sufficient to prime a TCF4-dependent rescuing of β-catenin nuclear activity that boost cell proliferation and invasiveness. Thus, let us suggest that our system could provide a mechanism-based rationale to further dissect key aspects of mechanotranscription in breast cancerogenesis and progression, somewhat independent of β-catenin's nuclear accumulation.

G-Quadruplex-Binding Small Molecule Induces Synthetic Lethality in Breast Cancer Cells by Inhibiting c-MYC and BCL2 Expression.

Herein, a prolinamide-derived peptidomimetic that preferentially binds to c-MYC and BCL2 G-quadruplexes present in the promoter regions of apoptosis-related genes (c-MYC and BCL2) over other DNA quadruplexes are described. Biological assays, such as real-time quantitative reverse transcription, western blot, dual luciferase, and small interfering RNA knockdown assays, indicate that the ligand triggers a synthetic lethal interaction by simultaneously inhibiting the expression of c-MYC and BCL2 genes through their promoter G-quadruplexes. The ligand shows antiproliferative activity in MCF-7 cells that overexpress both MYC and BCL2 genes, in comparison to cells that overexpress either of the two. Moreover, the ligand induces S-phase cell-cycle arrest, DNA damage, and apoptosis in MCF-7 cells.

CircularRNA abhydrolase domain containing 16A-microRNA-1237-5p-apoptosis associated tyrosine kinase axis regulates the apoptosis of michigan cancer foundation-7 in breast cancer cells

Objective To investigate the circular RNA abhydrolase domain containing 16A (circRNA ABHD16A)-microRNA (miRNA, miR)-1237-5p-Apoptosis associated tyrosine kinase (AATK) axis on apoptosis of breast cancer cell munggan cancer foundation-7 (MCF-7). Methods The correlation between ABHD16A and miR-1237-5p and miR-1237-5p and AATK was analyzed by CSCD and TargetScan online analysis. The luciferase assay was used to detect whether circRNA ABHD16A targets miR-1237-5p and whether miR-1237-5p targets AATK. The expression of apoptotic marker proteins in AATK and breast cancer cells MCF-7 was detected by Western blot after over-expression or knockdown of circRNA ABHD16A. Cell viability of breast cancer cells MCF-7 was analyzed by methyl thiazolyl tetrazolium (MTT) assay; staining by Annexin-V was performed by MTT assay. The level of apoptosis of breast cancer cells MCF-7 was measured. Student’s t-test analyzed two sets of one-way analysis of variance. Results circRNA ABHD16A targets miR-1237-5p; miR-1237-5p targets the 320-326 and 469-475 regions of the AATK 3’ UTR. When overexpressing ABHD16A, AATK expression increased significantly (t=15.295, P<0.01), apoptosis-related protein bcl-2 antagonist/killer 1 (bak), bcl-2 Associated X Apoptosis Regulator (bax) and cleaved Caspase-9 (cleaved-CAS9) expression levels increased significantly (t=7.403, 8.381, 21.583, P<0.01). The expression of bcl-2 Apoptosis Regulator (bcl-2) was significantly decreased (t=6.301, P<0.01), the activity of MCF7 cells was decreased (t=17.392, P<0.01), and the level of apoptosis was increased (t=8.491, P<0.05). When ABHD16A was knocked down, the expression of AATK was significantly decreased (t=9.290, P<0.01), and the expression levels of apoptosis-related proteins bak, bax and cleaved-CAS9 were significantly decreased (t=8.381, 5.296, 11.492, P<0.01), while the expression of bcl-2 increased significantly (t=5.609, P<0.01), the activity level of MCF7 cells increased (t=5.694, P<0.01), and the level of apoptosis decreased (t=13.502, P<0.01). Conclusion circRNA ABHD16A targets miR-1237-5p and increases the expression of AATK and promotes the apoptosis of breast cancer cells MCF-7. Key words: CircularRNA abhydrolase domain containing 16A; MicroRNA-1237-5p; Apoptosis associated tyrosine kinase; Breast cancer; Michigan cancer foundation-7; Cell apoptosis

Evaluation of in vitro anticancer activity of wheat germ oil in MCF-7 breast cancer cell lines

Cancer is the main cause of death and morbidity worldwide. Among females, breast cancer is the most commonly diagnosed and the leading cause of cancer-related death. Despite an improvement in the treatment of cancer in the latest years, metastatic breast cancer treatment success is not at a desirable level. Plants have huge potential to endow us with noble drugs and current drugs of plant origin are important part of cancer chemotherapy. The aim of this study was to evaluate the antiproliferative activity of wheat germ oil in MCF-7 breast cancer cell line. Antiproliferative activity and apoptotic effect of wheat germ oil were examined using the XTT and flow cytometry assay, respectively. According to the XTT results, at concentrations greater than 0.5 mg/mL, wheat germ oil displayed significant antiproliferative activity in MCF-7 cells in a dose-dependent manner. The Annexin V binding assay results also demonstrated that the IC 50 concentration of wheat germ oil increased early and late apoptotic populations (p < 0.05). Though further studies are required to specifically identify compounds involved in the anticancer activity of wheat germ oil, our findings demonstrate that wheat germ oil inhibits proliferation of MCF-7 cells and its action is dose-dependent.

Cycloartane and Stigmastane Type Triterpenoids from Pothos scandens Inhibit Estradiol (E2) Induced Proliferations in Breast Cancer Cells

Four cycloartane type triterpenoids and three stigmastane type steroids were isolated from the methanolic extract of stem and root part of Pothos scandens L. (Araceae), a Bangladeshi medicinal plant by high performance liquid chromatographic technique. The compounds were characterized as 24-methylenecycloartanol (1), 24-methylenecycloartenone (2), 24-en-cycloartenone (3), 24-methylenecycloartanyl ferulate (4), stigmast-4-en-3-one (5), stigmast-4,22-diene-3-one (6) and β-sitosterol glucoside (7) through extensive 1D and 2D NMR spectroscopic studies. All the isolates were evaluated for their estrogenic/antiestrogenic activity using the estrogen-responsive breast cancer cell lines, MCF-7 and T47D. The results showed that all the compounds possess mild to strong antiestrogenic activity in both cell lines which was compared to positive control tamoxifen. 24- Methylenecycloartanol (1), which contains a hydroxyl group in its C-3 position, inhibited 90% of estradiol (E2)- induced cell proliferation in MCF-7 and T47D cell lines at a concentration of 0.01 μM only. 24- Methylenecycloartanyl ferulate (4) and stigmast-4,22-diene-3-one (6) showed 90% of estradiol (E2)-induced cell proliferation in T47D cell only at a concentration of 0.01 μM whereas 10.0 μM was required for 24- methylenecycloartanyl ferulate (4) for the same activity in MCF-7 cells. This is the first report of isolation of these compounds from the plant along with their antiestrogenic property.&#x0D; Dhaka Univ. J. Pharm. Sci. 18(1): 93-102, 2019 (June)

The inhibitory effect of Andrographis paniculata extract on proliferation of breast cancer cell line

Aerobic glycolysis is commonly used as the main energy source for cancer cell growth. Adenylate kinase 2 (AK2) plays an important role in maintenance of ATP production in the mitochondria. Neoandrographolide in Andrographis paniculata potentially inhibit AK2 activity in silico. This study aimed to investigate the inhibitory effect of A. paniculata extract on proliferation of breast cancer cell line. This experimental laboratory used MCF-7 cell line and A. paniculata plants, which were extracted using the maceration method with 70% ethanol. Neoandrographolide concentration in A. paniculata extract was determined using HPLC. A total 1x104 MCF-7 cells were incubated for 24 hours with or without 0.4% (v/v) dimethyl sulfoxide as solvent (SC) or negative control (NC) group. The same amount of MCF-7 cells was treated with A. paniculata extract with 3 different dosages: 222 ppm (AE1), 111 ppm (AE2) and 55.5 ppm (AE3). The inhibition of cell proliferation used the 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazolium bromide (MTT) assay. Data were analyzed using ANOVA and LSD tests with p value <0.05.Ethanol extract of A. paniculata contained 4.43 µM neoandrographolide. The inhibition of MCF-7 cell proliferation was found in AE1 (47.98%) and AE2 (30.50%) groups and was significantly different from the NC group. While the percentage of MCF-7 cell inhibition in the AE1 group differed from NC (p = 0.002) and SC (p = 0.013) groups. Ethanol extract of A. paniculata can inhibit MCF-7 cell proliferation. Further Study is needed to investigate the role of neoandrographolide on inhibition of AK2 activity in MCF-7 cells.

Synthesis of PEGylated methotrexate conjugated with a novel CPP6, in sillico structural insights and activity in MCF-7 cells

Currently, most of the available chemotherapy has a narrow therapeutic window. This compromises the use of high-dose treatments in patients with advanced-stage cancers, who often need innovative strategies the most. Here, we present the conjugation of methotrexate with a novel cell-penetrating hexa-peptide (CPP6), a GFLG enzymatically cleavable spacer, and a short-length polyethylene glycol. The modifications added to methotrexate were rationalized to improve the drug bioavailability, specificity, and cell entry. Our final conjugate, CPGM, was successfully synthesized in solid phase and in vitro cell viability assays have shown that this conjugated form of methotrexate retains cytostatic effect towards MCF-7 breast cancer cells.

Copper(II) complexes of coumarin-derived Schiff base ligands: Pro- or antioxidant activity in MCF-7 cells?

A series of copper(II) complexes of Schiff base-derived ligands (1–7) were studied for their pro- and antioxidant behaviour in the MCF-7 human breast cancer cell line. The coordination modes of two of the copper(II) complexes were investigated by pH-potentiometry, EPR and UV–Vis spectroscopic methods. The solution studies indicated that monomeric species are present in the Cu(II) – L1 system at neutral pH, whereas dinuclear species were observed in the case of the Cu(II) – L7 system. This difference in speciation was reflected in their relative cytotoxicities with the copper(II) complex of L1, showing significant cytotoxicity against MCF-7 cells whilst the complex of L7 was inactive. In fact, only three of the seven complexes studied in this series were cytotoxic to MCF-7 cells but this cytotoxicity did not correlate with their ability to bind to DNA, cleave DNA or act as a pro-oxidant. In contrast to previous copper(II) complexes studied by our group, the compounds studied here do not appear to lead to intracellular reactive oxygen species generation at any significant level. In a yeast-based assay, all of the copper complexes had the ability to protect Saccharomyces cerevisiae against menadione-induced oxidative stress but not hydrogen peroxide-induced stress, indicating a lack of catalase activity. Given that the adaptive mechanisms induced by hypoxia in cancer cells have selective effects, with a fine-tuned protection against damage and stress of many kinds, particularly against oxidative stress, chemotherapeutic compounds which are not pro-oxidants may offer a therapeutic advantage.

An insight into the anticancer mechanism of Tribulus terrestris extracts on human breast cancer cells.

Tribulus terrestris (TT), a herb belonging to Zygophyllaceae family is widely used due to its medicinal properties. This study was undertaken to elucidate the anticancer mechanism of TT on MCF-7 breast cancer cells. Cytotoxic effect of the herb was assessed by 3-(4,5-diethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Apoptotic potential was assessed through DNA fragmentation, TUNEL and caspase 3 activity assays. Expressions of genes regulating the apoptotic pathway were examined by RT-PCR and expression of proteins was analyzed by immunocytochemistry. The result of MTT assay revealed that methanolic and saponin extracts from leaves and seeds of TT were cytotoxic to MCF-7 cells. Cytotoxicity studies on peripheral blood mononuclear cells (PBMC) proved that TT extracts were non-toxic to non-malignant cells. Treatment of human breast cancer MCF-7 cells with seed and leaf methanol andsaponin extracts of TT resulted in fragmentation of DNA and induction of apoptosis. This was evident by agarose gel electrophoresis of DNA and TUNNEL assay. The extracts of TT also caused a significant increase in caspase 3 activity in MCF-7 cells. TT extracts caused an induction of intrinsic apoptotic pathway which was evident by the upregulation in the expression of Bax and p53 genes and downregulation in the expression of Bcl-2. FADD, AIF and caspase 8 genes were also upregulated indicating the possible induction of extrinsic apoptotic pathway. Therefore, our results suggest that the Tribulus terrestris (TT) extracts may exert their anticancer activity by both extrinsic and intrinsic apoptotic pathways.

EVALUATION OF ANTIOXIDANT AND CHEMOPREVENTIVE POTENTIAL OF METHANOLIC EXTRACTS OF LEAF OF AEGLEMARMELOS ATTRIBUTES TOWARDS DUCTAL CARCINOMA STUDIED IN MCF7 CELLS

Objective: The objective of the present research work had been made to evaluate the antioxidant potential along with anti-cancer activity of methanolic extracts from the leaf of A. marmelos.&#x0D; Methods: Standard methods for antioxidant potential in terms of DPPH and nitric oxide scavenging assay, and anticancer activity in vitro method (Cytotoxicity/MTT assay and % of cell viability) by using MCF7 cell line.&#x0D; Results: Results of antioxidant efficacy revealed that the IC50 value for DPPH and nitric oxide scavenging assay was considered to be 62.032%, and 20.69% respectively. The methanolic extract of A. marmelos was found to possess enhanced anticancer potential against MCF7 cells. Cytotoxicity activity of MCF7 cells, when treated with methanolic extract of A. marmelos, was found to be 43.42% at 25µg, 52.31% at 50µg, 56.31% at 75µg, 58.38% at 100µg, 62.25% at 125µg. The IC50 value was found as 49.36µg. Toxicity was significantly increased with increased concentration and viability significantly decreased with the increased concentration of methanolic extract of leaf from A. marmelos for MCF7 cell when compared to cyclophosphamide.&#x0D; Conclusion: From the studies, it was postulated that methanolic extract of leaf from A. marmeloshas significant chemopreventive activity. These specific identities will be useful for the identification and authentication of raw drug.