Treatment Of MCF-7 Cells Research Articles

Effect of hydrophobicity on distinct anticancer mechanism of antimicrobial peptide chensinin-1b and its lipoanalog PA-C1b in breast cancer cells.

Chensinin-1b and its lipoanalogs demonstrate different anticancer activities against selected cancer cells, and the anticancer activity of PA-C1b is improved up to 3-fold compared with that of the parent peptide chensinin-1b. However, detailing the mechanism of action of these peptides is required to better understand the structure-function relationship. In this study, chensinin-1b and PA-C1b were selected as the representative peptides to investigate the mode of action in cancer cells. The results indicated that the boundary of the cell membrane was broken when the cells were treated with chensinin-1b, while that of cells treated with PA-C1b remained intact based on morphological changes. Apoptosis assays indicated that PA-C1b induced MCF-7 cancer cell apoptosis, while chensinin-1b mainly damaged the cell membrane. MCF-7 cancer cells treated with the peptides induced the loss of mitochondrial membrane potential, and cytochrome c was released from mitochondria, but PA-C1b enhanced ROS generation. Additionally, PA-C1b uptake occurred via an energy-dependent pathway and was inhibited by selected endocytosis inhibitors. Furthermore, treatment of MCF-7 cells with PA-C1b suppressed Bcl-2 mRNA levels and increased Bax mRNA levels, upregulated the expression of the proapoptotic protein Bax and downregulated the expression of the antiapoptotic protein Bcl-2. These results indicate that the anticancer mechanism of AMPs may be considerably affected by only a slight difference in the hydrophobicity of the two peptides; and such a study may facilitate the design of novel peptide-based anticancer agents.

Barrientosiimonas humi ethyl acetate extract exerts cytotoxicity against MCF-7 and MDA-MB-231 cells via induction of apoptosis and cell cycle arrest

Objective: To elucidate the cytotoxic effect of the secondary metabolites of Barrientosiimonas humi (B. humi) on MCF-7 and MDA-MB-231 human breast cancer cells and its underlying mechanisms of action. Methods: The extract was obtained from the fermentation of B. humi and fractionation of the crude extract was conducted via column chromatography. Cytotoxicity of the B. humi extract was determined by using MTT assay and real-time cellular analysis. Morphological changes, cell cycle profiles, mode of cell death, and caspase expressions of control and treated breast cancer cells were determined. Results: The ethyl acetate extract isolated from B. humi was cytotoxic against MCF-7 and MDA-MB-231 cell lines. One of the dichloromethane (DCM) fractions, designated as DCM-F2, exhibited the strongest activity among all the fractions and thereby was selected for further studies. DCM-F2 had selective cytotoxicity on target cells by inducing apoptosis, particularly in the early stage, and cell cycle arrest. Treated cells caused inhibition of cell cycle progression at 72 h leading to a significant increase (P < 0.05) in the G0/G1 population. DCM-F2 treated MDA-MB-231 cells showed caspase-dependent apoptosis, whereas DCM-F2 treated MCF-7 cells showed a caspase-independent apoptosis pathway. Five compounds were successfully isolated from B. humi. Cyclo (Pro-Tyr) was the most cytotoxic and selective compound against MCF-7 cells. Conclusions: B. humi ethyl acetate extract exhibits significant cytotoxicity against MCF-7 and MDA-MB-231 cells via induction of apoptosis and cell cycle arrest.

Differential Expression of Platelet Activation Markers, CD62P and CD63, after Exposure to Breast Cancer Cells Treated with Kigelia Africana, Ximenia Caffra and Mimusops Zeyheri Seed Oils In Vitro

Cancer patients, including breast cancer patients, live in a hypercoagulable state. Chemo- and hormone- therapy used in the treatment of breast cancer increases the risk of thrombosis. Due to differences in health care services between developed and developing countries, the survival rate of women with breast cancer in developing countries is low. Consequently, ethnomedicines are used and their efficacy as potential alternatives are being scientifically explored. The seed oils of Kigelia africana, Ximenia caffra and Mimusops zeyheri have anti-proliferative effects on hormone-dependent (MCF-7) and cytotoxic effects on hormone-independent (MDA-MB-231) breast cancer cells. In this study, we determined if these seed oils reduce the thrombogenic ability of breast cancer cells by measuring the platelet surface expression of the activation-specific antigens CD62P and CD63. MDA-MB-231 and MCF-7 cells were pretreated with the seed oils before being exposed to whole blood of human female volunteers. An increase in CD62P and CD63 expression following whole blood exposure to untreated breast cancer cells was observed. Treated MDA-MB-231 cells reduced CD62P and CD63 expression while treated MCF-7 cells increased CD62P and decreased CD63 expression. Kigelia africana, Ximenia caffra and Mimusops zeyheri seed oils are able to reduce the thrombogenic ability of MDA-MB-231 breast cancer cells.

Design and Activity of Novel Oxadiazole Based Compounds That Target Poly(ADP-ribose) Polymerase.

Novel PARP inhibitors with selective mode-of-action have been approved for clinical use. Herein, oxadiazole based ligands that are predicted to target PARP-1 have been synthesized and screened for the loss of cell viability in mammary carcinoma cells, wherein seven compounds were observed to possess significant IC50 values in the range of 1.4 to 25 µM. Furthermore, compound 5u, inhibited the viability of MCF-7 cells with an IC50 value of 1.4µM, when compared to Olaparib (IC50 = 3.2 µM). Compound 5s also decreased cell viability in MCF-7 and MDA-MB-231 cells with IC50 values of 15.3 and 19.2 µM, respectively. Treatment of MCF-7 cells with compounds 5u and 5s produced PARP cleavage, H2AX phosphorylation and CASPASE-3 activation comparable to that observed with Olaparib. Compounds 5u and 5s also decreased foci-formation and 3D Matrigel growth of MCF-7 cells equivalent to or greater than that observed with Olaparib. Finally, in silico analysis demonstrated binding of compound 5s towardsthe catalytic site of PARP-1, indicating that these novel oxadiazoles synthesized herein may serve as exemplars for the development of new therapeutics in cancer.

Effects of Endocrine Disruptors o,p'-Dichlorodiphenyltrichloroethane, p,p'-Dichlorodiphenyltrichloroethane, and Endosulfan on the Expression of Estradiol-, Progesterone-, and Testosterone-Responsive MicroRNAs and Their Target Genes in MCF-7 Cells.

Many studies have shown that dichlorodiphenyltrichloroethane (DDT) exposure raises breast cancer risk. Another insecticide with similar properties is endosulfan, which has been actively used in agriculture after DDT prohibition. Previously, we have identified some estradiol-, progesterone-, and testosterone-sensitive microRNAs (miRNAs, miRs). Because DDT and endosulfan have estrogenic, antiandrogenic, and antiprogesterone properties, we hypothesized that these miRNAs are affected by the insecticides. We quantified relative levels of miRNAs and expression levels of their target genes in breast cancer MCF-7 cells treated with p,p′-DDT, o,p′-DDT, or endosulfan. We also quantified miR-19b expression, which, as previously shown, is regulated by estrogen. Here, we observed that miR-19b expression increased in response not only to estradiol but also to testosterone and progesterone. Treatment of MCF-7 cells with p,p′-DDT or endosulfan decreased the protein levels of apoptosis regulators TP53INP1 and APAF1. In cells treated with o,p′-DDT, the TP53INP1 amount decreased after 24 h of incubation, but increased after 48 h of incubation with insecticide. OXTR expression, which is known to be associated with breast carcinogenesis, significantly diminished under the exposure of all insecticides. In cells treated with p,p′-DDT or o,p′-DDT, the observed changes were accompanied by alterations of the levels of hormone-responsive miRNAs: miR-324, miR-190a, miR-190b, miR-27a, miR-193b, and miR-19b.

Biogenic synthesis of gold nanoparticles mediated by Spondias dulcis (Anacardiaceae) peel extract and its cytotoxic activity in human breast cancer cell

Green synthesis is a new paradigm for the preparation of gold nanoparticles (AuNPs) due to its cost-effectiveness and favorable environmental impact. This study presented a simple phytosynthesis process for the preparation of AuNPs utilizing the aqueous peel extract of Spondias dulcis (SPE) (Anacardiaceae) as both a reducing and stabilizing agent. A visual color change from yellow to purple during the reaction implied the successful formation of SPE-AuNPs, which was confirmed by UV–vis spectroscopy. Transmission electron microscopy (TEM) images indicated that the SPE-AuNPs were predominantly spherical with a mean size of 36.75 ± 11.36 nm, and were comprised of crystalline Au, as indicated by X-ray diffraction. In terms of their potential application, SPE-AuNPs exhibit significant cytotoxic activity in a dose- and time-dependent manner to MCF-7 human breast cancer cells, while being non-toxic to Vero normal cells. The treatment of MCF-7 cells with SPE-AuNPs increased the production of intracellular reactive oxygen species (ROS). Herein, the findings highlight the potential contribution of phytosynthesized SPE-AuNPs to the development of novel nanomedicines for cancer treatment.

Sinigrin Impedes the Breast Cancer Cell Growth through the Inhibition of PI3K/AKT/mTOR Phosphorylation-Mediated Cell Cycle Arrest.

Breast carcinoma, one of the most lethal variants of carcinogenesis, significantly diagnosed type of cancer amongst the female population. Sinigrin, also known as glucosinolate, is found in the seeds of Brassica nigra and shown to enhance various cancer cells potentially. Nevertheless, the mechanistic explanation of sinigrin (SGN)-mediated breast cancer growth and augmentation is still to be investigated. Therefore, we contended in this study that SGN impedes PI3K/AKT/mTOR phosphorylation-mediated cell cycle arrest in MCF-7 cells. SGN (20 M) was implemented to treat MCF-7 cells for 24 and 48 hours of incubation. A significant increase in cytotoxicity, reactive oxygen species (ROS) generation, cell cycle arrest, mitochondrion membrane alteration, lipid peroxidation, and antioxidant depletion was found in MCF-7 cells. The PI3K/AKT/mTOR events are crucial pathways that participate in survival, proliferation, and cell cycle regulation. Inhibition of PI3K/AKT/mTOR expression thought to be novel approach for alleviating breast cancer growth. We noticed that SGN inhibits PI3K, AKT, and mTOR phosphorylation, resulting in the downregulation of proliferative and cell cycle regulatory proteins, such as cyclin-Dl, PCNA, CDK4, and CDK6. SGN also causes apoptosis in MCF-7 cells by increasing nuclear fragmentation and by inducing pro-apoptotic gene expression. As a result, SGN inhibits breast cancer growth by impeding PI3K/AKT/mTOR phosphorylation-mediated cell cycle arrest in MCF-7 cells.

Morphological damage and increased ROS production of biosynthesized silver nanoparticle against MCF-7 breast cancer cells through in vitro approaches

In this study of anti-cancer activity, the cytotoxicity effect of the silver nanoparticle was shown excellent anti-cancer activity against MCF-7 breast cancer cells. The IC50 concentration of the synthesized silver nanoparticle against performed MCF-7 breast cancer cells was 1000 µg/ml concentration. Further, the shape modifications in the silver nanoparticle treated MCF-7 cells were viewed by phase contrast microscope and proved after comparison with untreated control. Next, the intracellular proliferation, chromatin condensation, granular membranes cleavage and surface layers damage were effectively seen by fluorescence microscopy using AO/EB fluorescence dyes. The failure of matured nucleus was clearly shown after the treatment of Ag NPs and was effectively proved by fluorescence microscope using Hoechst 33342 stain. Finally, the anti-cancer effect identification through various invitro experiments was clearly favored to performed silver nanoparticle, and suggested that the nanoparticle was very effective against multi drug resistant bacteria, biofilm producing bacteria and cancer cells.

Fluorizoline Blocks the Interaction between Prohibitin-2 and γ-Glutamylcyclotransferase and Induces p21Waf1/Cip1 Expression in MCF7 Breast Cancer Cells

Prohibitin-2 (PHB2) is a scaffold protein that has pleiotropic functions, which include interacting with γ-glutamylcyclotransferase (GGCT) in the cytoplasm and repressing the transcriptional activities of the p21Waf1/Cip (p21) gene in the nucleus. The cytotoxic drug fluorizoline binds to PHB1/2 and exerts antiproliferative actions on cancer cells. However, the precise mechanism underlying the antiproliferative effects of fluorizoline is not fully elucidated. In the present study, we first show that fluorizoline induces p21 expression in several human cancer cell lines, including MCF7 breast cancer cells. Treatment of MCF7 cells with fluorizoline suppressed proliferation and prevented cells from entering into the DNA synthesis phase. Knockdown of p21 rescued the suppressed proliferation, indicating that fluorizoline inhibited MCF7 cell growth via the induction of p21. Overexpression of PHB2 in MCF7 cells prevented the induction of p21 expression by fluorizoline and restored the antiproliferative effects and blockade of cell cycle progression. Moreover, treatment of MCF7 cells with fluorizoline inhibited the interaction between endogenous PHB2 and GGCT proteins and reduced the level of nuclear localization of PHB2 proteins. These results indicate that targeting PHB2 with fluorizoline induces the expression of p21 and consequently blocks proliferation of cancer cells. SIGNIFICANCE STATEMENT: This study shows that fluorizoline may be a promising novel anticancer drug candidate that induces p21 expression and blocks cell-cycle progression in human cancer cell lines. In addition, we show that fluorizoline inhibits the interaction between PHB2 and GGCT and reduces the nuclear localization of PHB2 proteins.

Carbonic anhydrase 12 gene silencing reverses the sensitivity of paclitaxel in drug-resistant breast cancer cells

ABSTRACT This study aimed to investigate the effects of carbonic anhydrase 12 (CA12)-siRNA on the paclitaxel sensitivity of breast cancer cells. Normal mammary glandular cell (MCF-10), breast cancer cell (MCF-7), and paclitaxel-resistant breast cancer cells (MCF-7 TaxR) were cultured in experimental control group. Western blot was adopted to detect the expressions of CA12 protein and apoptosis-related proteins in mitochondrial pathway of MCF-10, MCF-7, and MCF-7 TaxR cells. The methylthialazole tetrazolium (MTT) method was used to measure cell proliferation. The apoptosis of MCF-7 and MCF-7 TaxR cells was observed in phase contrast microscope, fluorescence inverted phase contrast microscope, and flow cytometry (FACS). The results showed that CA12 protein expression in MCF-7 and MCF-7 TaxR cells was significantly higher than that in MCF-10 cell. The growth rate of CA12-siRNA treated MCF-7 TaxR cells with paclitaxel (PTX) co-culture was markedly declined at 48 hours. Phase contrast microscope, fluorescence inverted phase contrast microscope, and FACS showed that apoptotic cells in the CA12-siRNA treated MCF-7 TaxR groups were significantly higher than that in CA12-siRNA treated MCF-7 cells. The expressions of pro-apoptotic proteins, Bax and Bid, were dramatically increased in CA12 siRNA treated MCF-7 TaxR cells. The expression quantity of the downstream effective molecules caspase-9, caspase-7, and the activated proteins of poly (ADP-ribose) polymerase (PARP), also were significantly increased. Our results indicated that the application of PTX combined silencing CA12 was able to activate the mitochondrial apoptosis pathway and promote MCF-7 TaxR apoptosis. CA12 silencing in the PTX-resistant breast cancer cell can reverse the sensitivity of PTX.

Regulation of Chloride Intracellular Channel Protein 1 and Caspase -3 mRNA Expression by Hydroethanolic Extract of Aegle marmelos Fruit Human Breast Cancer Cell Line-MCF-7

Introduction: Cancer is the second leading cause of death all over the world where among all types of cancer breast cancer is said to be the leading cancer followed by lung cancer. The aim of this study is to find the regulation of chloride intracellular channel protein 1 and caspase -3 mRNA expression by hydroethanolic extract of Aegle marmelos fruit human breast cancer cell line-MCF-7.
 Materials and methods: MCF-7 cells were collected from NCCS Pune, India. It is stored in Dubecos Modified Eagle's Medium. The Aegle marmelos fruit was collected from the herbal department and its extract was prepared. The extract of Aegle marmelosis used in treating MCF-7 cells at different dosages in in vitro. Isolation of total RNA from MCF-7 cells. The cells will be mixed with total RNA isolation reagent, sonicated and RNA will be isolated as per the standard method. c-DNA conversion and real time polymerase chain reaction. The c-DNA will be synthesized using reverse transcription by commercially available (RT-PCR) kit. Two microlitres of c-DNA will be used for amplification of clic-1 and caspase-3 using gene specific primers by commercially available RT-PCR kit (SyBr kit) and comparative CT method will be used to see the expression of genes. Untreated MCF-7 cells were compared with MCF-7 cells treated with various concentrations of the extract (10, 20 and 40ug). The statistical data’s were collected from the SPSS software version 21.
 Result: The given extract inhibits the proliferation of MCF-7 cells therefore said to have antiproliferative activity. Different doses of extract were tested (200ug-500ug) out of which 400ug of extract were preferred.
 Conclusion: The given plant extract has anti proliferative properties and hence can be used as a drug to treat breast cancer.

Christia vespertilionis extract induced antiproliferation and apoptosis in breast cancer (MCF7) cells.

C. vespertiliomis extracts were evaluated for antiproliferative and apoptosis effect on breast cancer (MCF7) cells. The leaves extracts were analysed for its antiproliferative effect on breast cancer (MCF7) cells and normal epithelial breast (MCF 10A) cells using Sulforhodamine B (SRB) assay. The selective extract was evaluated for its ability to induce apoptosis using Annexin V-FITC apoptosis staining and the expression of molecular genes using qualitative reverse transcription-polymerase chain reaction (RT-PCR) against MCF7 cells. Gas chromatography-mass spectrometry (GC-MS) was used to identify the compounds from the selective extract. The findings showed that dichloromethane fraction (CV-Dcm) extract had high antiproliferative effect against MCF7 cells (IC50 = 24µg/mL, selective index (SI) = 8.17). The percentages of apoptosis cells in CV-Dcm-treated MCF7 cells was 58.8%. The CV-Dcm extract induced downregulation of PCNA level. The apoptotic genes were also triggered in both extrinsic and intrinsic signaling pathways, affecting a 1.5-fold increase in BAX, 1.4-fold increase in cytochrome c, 1.3-fold increase in caspase-8, 1.7-fold increase in caspase-3 and 0.5-fold-decrease in BCL-2. Treated MCF7 cells also activated P53-dependent apoptotic death pathway. The present work strongly suggests that high efficacy of CV-Dcm extract was attributed to its antiproliferative and apoptosis-inducing activation in MCF7 cells, most likely due to its favourable compounds.

Expression of Phosphatase and Tensin Homolog Deleted on Chromosome Ten on the Development of Cisplatin Resistance in Breast Cancer by TGF-β1 Signal Pathway

Introduction: This study aims to evaluate the expression of PTEN on the development of cisplatin resistance in breast cancer by TGF-β1 signal pathway in order to investigate the prognostic value of PTEN and TGF-β1 for breast cancer. Material and Methods: In this study, HE staining, Immunohistochemical staining, Immunofluorescence and Western blotting were used to detect the expression of PTEN and TGF-β1 in breast cancer MCF-7 cells to explain clearly the relationship between PTEN and TGF-β1 on TGF-β1 treated MCF-7 Cells. Results: The experiment results showed the expression of PTEN and TGF-β1 in Human breast cancer increased obviously compared with paracancerous tissues (P < 0.05). The expression of PTEN and TGF-β1 was closely correlated with tumor size, distant metastasis, pathological stage and progesterone receptor status, but not with age and lymph node status. At the same time, the expression of PTEN and TGF-β1 in cisplatin-treated MCF-7 Cells reduced as compared with untreated breast cancer cells (P < 0.05). In addition, the expression of PTEN increased by TGF-β1 inducer treated Human Breast Cancer Cells. However, the expression of PTEN in TGF-β1 inhibitor treated Human Breast Cancer Cells was lower than in untreated breast cancer cells (P < 0.05). Conclusions: PTEN and TGF-β1 played an important role in Human Breast cancer. In addition, the expression of PTEN could be regulated by TGF-β1 in Cisplatin and TGF-β1 treated Human Breast Cancer Cells.

Nanoencapsulation of Hirudo medicinalis proteins in liposomes as a nanocarrier for inhibiting angiogenesis through targeting VEGFA in the Breast cancer cell line (MCF-7).

Introduction: Breast cancer is the most serious cause of women’s death throughout the world. Using nanocarrier vehicles to the exact site of cancer upgrades the therapeutic efficiency of the drugs. Capsulation of active proteins in the vesicular liposomes’ hydrophilic core is essential to develop a therapeutic protein carrier system. We aimed to encapsulate the medicinal leech saliva extract (LSE) and assess the inhibition of angiogenesis of breast cancer cells by targeting vascular endothelial growth factor A (VEGFA). Methods: In this research, enhanced formulation of liposomal protein was determined by zeta potential analysis, droplet size, drug release assay, and transmission electron microscopy (TEM). Furthermore, a cytotoxicity assay of liposomal LSE was performed to determine the cytotoxic activity of components. For assessing the expression of VEGFA, P53, and hypoxia-inducible factor subunit alpha (HIF1a) genes, Real-Time PCR was applied. Results: Nano liposome was chosen as an enhanced formulation due to its much smaller size (46.23 nm). Liposomal LSE had more practical actions on the MCF-7 cells. As noticed by DAPI staining, apoptosis was extensively greater in treated MCF-7 cells. Wound healing assay demonstrated that MCF-7 cells could not sustain growth at the presence of liposomal LSE and expression of the VEGFA gene was declined in treated cells. Downregulation of VEGFA was evaluated with western blotting technique. Conclusion: It can be concluded that our investigation of the tests confirmed the fact that nano liposomal LSE is a novel promising formulation for anticancer drugs and can significantly improve the penetration of protein drugs to cancer cells.

The Cytotoxic effect of an Ethanol extract of Momordica Charantia, Kuguacin-J and Cisplatin on healthy MCF-10A and MCF-7 and MDAMB-231 breast cancer cell lines employing In Vitro assays

Traditional medicines, derived from plants, could present alternative treatment strategy for cancer therapy. One such plant is Momordica charantia (MC) which possesses anti-carcinogenic properties. This study investigated the anticancer effect of an ethanol extract of MC fruit, Kuguacin-J (K-J), an isolated compound from the leaves of MC and cisplatin, either alone or combination on healthy MCF-10A mammary cells and compared with breast cancer MCF-7 and MDAMB-231 cell lines. Cell viability was tested using 8 μg/mL and 80 μg/mL doses of MC extract, K-J and cisplatin individually or combined for 24 and 48 hours. Caspase-3- activity was measured in MCF-7 and MDA-MB-231 cells using established methods. The results revealed that MC extract and K-J had no effect on healthy MCF-10A cell viability as compared to cisplatin which induced dose and time-dependent cell death. Similarly, treatment of MCF-7 cells with cisplatin induced cell death at high concentration at both the time points, while MC extract and K-J only induce MCF-7 cell death at high dose after 48 hours only. During combination therapy, both doses of cisplatin enhanced MCF-7 cell death when combined with MC extract or K-J after 24 and 48 hours. In MDAMB-231 cells, the three drugs, either alone or combined, evoked significant cell death at both the doses and time points. All three drugs, at high dose, elicited significant increase in caspase-3- activity in MCF-7 and MDA-MB-231 cells as compared to untreated cells. The results revealed that either MC extract or K-J alone or combined with cisplatin, can elicit significant increase in cell death and caspase–3-activity in MCF-7 and MDA-MB-231cells as compared to untreated cells.

Chemical Composition, α-Glucosidase Inhibitory and Anticancer Activity of Essential Oil of Thymus vulgaris Leaves

Dietary herbs are used for imparting flavour, aroma, and piquancy to foods. Besides their role in improving food quality, spices also have health benefits, such as anticancer, antidiabetic, antimicrobial, antioxidant, hypolipidemic, analgesic, immunostimulant, etc. Thymus vulgaris (family: Lamiaceae) is a valuable medicinal herb known for its health benefits. The essential oil from T. vulgaris was isolated by the hydrodistillation method, and chemical constituents in it were determined by GC-MS. Anticancer activity was determined using a DNA ladder assay, and antidiabetic activity was accessed by α-glucosidase inhibition. The GC-MS analysis showed the presence of 16 chemical constituents, and thymol (55.88 %), linalool (13.71 %), carvacrol (8.36 %), and p-cymene (6.00 %) were found as major components. The antidiabetic activity of T. vulgaris essential oil showed potent inhibition of α-glucosidase enzyme with an IC50 value 125.1±4.25 μg/mL. The anticancer activity results revealed that treating MCF-7 cells with 100 μg/mL essential oil of thyme produces DNA fragmentation and promotes apoptosis. The current study provides a scientific basis for its possible use in controlling diabetes and breast cancer. The study further expands the knowledge of T. vulgaris chemical composition and provides evidence of ethnomedicinal use of the plant by exploring antidiabetic and anticancer activities.

Abstract 2637: The use of 3D-printed Polylactic acid scaffolds (PLA) as a substrate for growth of MCF7 human breast cancer cells

Abstract This study focuses on fabrication of large-scale tissue scaffolds from Polylactic acid (PLA) and their feasibility as a substrate for cell growth. The scaffolds were fabricated via combined additive manufacturing and microwave foaming. The benefit of this approach is the ability to generate scaffolds with specific architectures and controllable pore size and porosity. The resulting scaffolds have dual pore networks with porosity in excess of 80% which provide a pathway for both cell growth and circulation of culture media. We sought to determine the capability of these scaffolds to support attachment, growth and viability of the MCF7 human breast cancer cell line, as well as drug sensitivity. For these experiments the PLA scaffolds were sterilized in 24-well plates and 1 x 105 MCF7 cells were seeded into each well and cultured for up to 7 days. Scaffolds were then used for SEM imaging, hoescht-staining, or cell counting and viability measurements after trypsinization. Comparisons of cell numbers detached from the scaffolds vs. the remaining well suggest a proportionally small percentage of cells (less than 5%) attached to the PLA, which is not surprising since the scaffolds lack a solid bottom. However, our imaging data using hoescht-staining of PLA-attached cells pre- and post- trypsinization suggest that traditional trypsinization does not effectively detach cells from the PLA material, and therefore this method cannot be used to accurately determine the proportion of cells growing on the scaffolds. This also reveals a potential limitation for application. SEM imaging confirmed cell attachment, with cell growth in the form of sheets along the struts on the bottom layer, and clusters elsewhere with a size of around 50 micron. Analysis of viability using trypan blue exclusion shows no effect of the PLA material on cell viability. To determine the drug sensitivity of cells grown with the PLA material, we treated MCF7 cells grown in the presence or absence of oleuropein, the main phenolic compound found in olive leaves. Media was collected at 48 hours and 7 days and released LDH measured by a cytotoxicity assay. Our data showed that the PLA material itself did not increase cellular toxicity in the absence of oleuropein treatment. It also confirmed that treatment with 200 ug/ml oleuropein induces the expected cytotoxicity in MCF7 cells grown in the absence of PLA, with no significant difference in oleuropein-susceptibility for cells grown in the presence of PLA scaffolds. Together, our results suggest that PLA scaffolds can support MCF7 cell attachment and growth, with no effect on cell viability or drug-induced cytotoxicity. This provides support for the promise of PLA scaffolds for physiologically relevant cell and tissue culture work. Citation Format: Brenna McAllister, Mohammed Mustafa, Sriharsha Sundarram, Shelley A. Phelan. The use of 3D-printed Polylactic acid scaffolds (PLA) as a substrate for growth of MCF7 human breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2637.

Mixed phytochemicals mediated synthesis of copper nanoparticles for anticancer and larvicidal applications

The synthesis of copper nanoparticles (CuNPs) using Wrightia tinctoria (Wt) R.Br extract is defined in this article as being convenient, environmentally friendly, and non-toxic. UV-visible spectrophotometry, FT-IR, XRD, particle size analyser, SEM-EDAX and TEM methods were used to describe the physicochemical properties of Wt extract mediated synthesized CuNPs (Wt-CuNPs). The Wt-CuNPs synthesized was found to be monodispersed and spherical, with an average size of 15 nm. Gas chromatography and mass spectrometry (GC-MS) research revealed that the Wt R.Br plant extract contains various phytochemical compounds. The properties of Wt-CuNPs were verified by the findings of characterization tests. Via in silico molecular docking experiments with established targets, the underlying mechanisms of cytotoxicity against breast cancer and larvicidal behaviour against Aedes aegypti of Wt-CuNPs were investigated. Interestingly, in vitro cytotoxicity studies showed 50% cell death (IC50) of Wt-CuNPs treated MCF-7 cells and Vero Cells (Kidney epithelial cells) were displayed at 119.23 μg.mL−1 and 898.75 μg.mL−1, respectively. Also, Wt-CuNPs showed least LC50 and LC90 values for larvicidal activity against A. aegypti were of 32.10 μg.mL−1 and 21.70 μg.mL−1, respectively. Furthermore, Wt-CuNPs is found to be less toxic and biocompatible in haemolytic assays. The findings clearly showed that biosynthesized Wt-CuNPs have been used as a possible anticancer and larvicidal agent, as well as being environmentally friendly.

Doxorubicin-paclitaxel sequential treatment: insights of DNA methylation and gene expression changes of luminal A and triple negative breast cancer cell lines.

Breast cancer is one of the significant causes of death among women diagnosed with cancer worldwide. Even though several chemotherapy combinations are still the primary treatment of breast cancer, unsuccessful treatments, and poor prognostic outcomes are still being reported. DNA methylation and gene expression changes among two breast cancer cell lines representing luminal A (MCF-7) and triple-negative (MDA-MB-231) cancers were determined after sequential combination treatment of doxorubicin and paclitaxel and analyzed using Ingenuity Pathway Analysis. Promoter methylation changes were seen in different treated MCF-7 cells and accompanied by changes in the gene expression of CCNA1 and PTGS2. In MDA-MB-231 cells, the hypomethylation of ESR1 was not accompanied by an increase in its gene expression in any treated cells. The hypomethylation of GSTP1 and MGMT was accompanied by an increase in gene expression levels in the group treated with doxorubicin only. Also, significant downregulation of several genes like MUC1 and MKI67 in MCF-7 cells treated with doxorubicin showed much lower gene expression (- 37.63, - 10.88 folds) when compared with cells treated with paclitaxel (- 2.47, - 2.05 folds) or the combination treatment (- 18.99, - 2.81 folds), respectively. On the other hand, a synergistic effect on MMP9 gene expression was significantly seen in MDA-MB-231 cells treated with the combination (- 9.99 folds) in comparison with the cells treated with doxorubicin (- 3.62 folds) or paclitaxel (1.75 folds) alone. Chemotherapy combinations do not always augment the molecular changes seen in each drug alone, and these changes could be utilized as treatment response markers.

Synthesis, docking study, and in vitro anticancer evaluation of new flufenamic acid derivatives

Novel compounds (6–10) were synthesized and confirmed by spectroscopic analysis, including AT-IR, 1HNMR and CHNS. Their cytotoxic effect was evaluated by MTT assay against two cancer cell lines and two normal cell types. Compound 7 exhibited anticancer activity against MCF-7 breast cancer cell line (GI50 = 63.9 µg/ml, 148 µM), without any effect against A549 lung cancer cells, or the normal cells. Compound 7 caused cytotoxicity in MCF-7 breast cancer cells by apoptotic cell death, as suggested by fragmented nuclei after DAPI staining and agarose gel electrophoresis. In addition, treating MCF-7 cells with compound 7 resulted in an increase in the level of caspase 9 mRNA level, and its activation. Moreover, compound 7-treated MCF-7 cells showed enhanced cytochrome c release from the mitochondria to the cytosol, signifying an induction of the intrinsic apoptotic pathway. Finally, compound 7 exhibited epidermal growth factor receptor (EGFR) kinase inhibitory activity at (EC50 = 0.13 µM), which was matched by molecular docking studies that showed compound 7 might be an important EGFR kinase inhibitor.