RESUMO
SUMMARY: This study evaluated the phytochemical screening, antioxidant capacity, and in vitro anticancer activities of four plants namely, Gypsophila capillaris, Anabasis lachnantha, Haloxylon salicornicum, and Horwoodia dicksoniae which belong to four different families: Caryophyllaceae, Amaranthaceae, Chenopodiaceae, Brassicaceae, respectively. The total phenolics, anthocyanins, saponins, total antioxidant capacity (TAC), and DPPH assays were determined by spectrophotometer. In vitro anticancer activity was assessed using two human cancer cell lines; hepatocellular carcinoma (HepG-2) and breast adenocarcinoma (MCF-7) to estimate the inhibition concentration 50 % (IC50). The results showed that H. dicksoniae has the highest concentrations of phenolics and saponins, while H. salicornicum has the highest DPPH. The highest concentration of TAC was found in G. capillaries. Among the tested extracts, G. capillaries and H. salicornicum have the potential activity against MCF-7 and HepG-2 cell lines in vitro. The content of polyphenols in G. capillaries was profiled by high-performance liquid chromatography (HPLC). The highest concentration among the phenolic compounds was chlorogenic (60.8 µg/ml) while the highest concentration among the flavonoid compounds was hesperidin (1444.92 µg/ml). In summary, G. capillaries and H. salicornicum extracts have potent anticancer activity against HepG-2 and MCF-7 cell lines.
Este estudio evaluó la detección fitoquímica, la capacidad antioxidante y las actividades anticancerígenas in vitro de cuatro plantas, Gypsophila capillaris, Anabasis lachnantha, Haloxylon salicornicum y Horwoodia dicksoniae, que pertenecen a cuatro familias diferentes: Caryophyllaceae, Amaranthaceae, Chenopodiaceae y Brassicaceae, respectivamente. Los ensayos de fenólicos totales, antocianinas, saponinas, capacidad antioxidante total (TAC) y DPPH se determinaron mediante espectrofotómetro. La actividad anticancerígena in vitro se evaluó utilizando dos líneas celulares de cáncer humano; carcinoma hepatocelular (HepG-2) y adenocarcinoma de mama (MCF- 7) para estimar la concentración de inhibición del 50 % (IC50). Los resultados indicaron que H. dicksoniae tiene las concentraciones más altas de fenólicos y saponinas, mientras que H. salicornicum tiene el DPPH más alto. La mayor concentración de TAC se encontró en G. capillaries. Entre los extractos probados, G. capillaries y H. salicornicum tienen actividad potencial contra líneas celulares MCF-7 y HepG-2 in vitro. El contenido de polifenoles en G. capillaries se perfiló mediante cromatografía líquida de alta resolución (HPLC). La concentración más alta entre los compuestos fenólicos fue clorogénica (60,8 µg/ml), mientras que la concentración más alta entre los compuestos flavonoides fue la hesperidina (1444,92 µg/ml). En resumen, los extractos de Gypsophila capillaris y H. salicornicum tienen una potente actividad anticancerígena contra las líneas celulares HepG-2 y MCF-7.
Assuntos
Humanos , Plantas Medicinais/química , Antineoplásicos/química , Antioxidantes/química , Fenóis/análise , Saponinas/análise , Arábia Saudita , Técnicas In Vitro , Cromatografia Líquida de Alta Pressão , Metabolômica , Células Hep G2/efeitos dos fármacos , Células MCF-7/efeitos dos fármacos , Compostos Fitoquímicos , Antocianinas/análise , Antineoplásicos/farmacologia , Antioxidantes/farmacologiaRESUMO
@#[摘 要] 目的:探究甘露糖结合凝集素2(LMAN2)在激素受体(HR)阳性乳腺癌组织中的表达水平与乳腺癌患者预后的关系及其对MCF-7 细胞增殖和迁移的影响。方法:通过TCGA、Bc-GenExMiner、GEPIA和Kaplan-Meier Plotter数据库分析LMAN2在乳腺癌组织和正常乳腺组织中的差异性表达及其与患者预后的关系。采用小RNA干扰技术将si-LMAN2#1、si-LMAN2#2及si-NC转染至MCF-7细胞,将过表达LMAN载体(pc-LMAN)及空载体pcDNA3.1阴性对照(pc-NC)转染至MCF-7细胞,实验分为si-LMAN2#1、si-LMAN2#2、si-NC、pc-LMAN2和pc-NC组。通过qPCR和WB实验检测各组细胞中LMAN2 mRNA和蛋白的表达水平,CCK-8、克隆形成、Transwell迁移、WB等实验检测敲低和过表达LMAN 2对MCF-7细胞增殖、克隆形成、迁移及AKT信号通路相关蛋白表达的影响。结果:LMAN2在乳腺癌组织中的表达水平显著高于正常乳腺组织(P<0.001)。HR阳性乳腺癌组织中LMAN2表达水平显著高于HR阴性乳腺癌组织(P<0.001);LMAN2高表达与HR阳性乳腺癌患者不良预后有关联。敲低LMAN2可显著降低MCF-7细胞的增殖和迁移能力(P<0.01或P<0.001),过表达LMAN2可显著提高MCF-7细胞的增殖和迁移能力(均P<0.001)。敲低LMAN2组MCF-7细胞中PTEN和P21蛋白表达水平均显著升高,p-AKT蛋白表达水平显著降低(均P<0.01)。结论:LMAN2在乳腺癌组织和HR阳性乳腺癌组织中高表达,且与不良预后有关联。LMAN2高表达与MCF-7细胞增殖和迁移有关联,其作用机制可能涉及AKT信号通路。
RESUMO
Annona muricata Linn. (Annonaceae) is a tropical plant with multiple beneficial health effects including anticancer properties. In breast cancer patients, overexpression of the HER2 oncoprotein corresponds to a poor prognosis, thus the main purpose of this study was to evaluate the cytotoxicity of ethanolic extracts from dried and fresh leaf of A. muricata on HER2+ breast cancer cells. MTT assays were performed and IC50 determined in HCC1954 (HER2+) cells, as well as in MCF7 (HER-) and peripheral blood mononuclear cells (PBMC) used as controls. Total polyphenol content evaluation and phytochemical screening were also performed. The cytotoxic effect of A. muricata extracts (125-1000 µg/mL) was dose-dependent and cell-type specific. The extracts exhibited higher cytotoxicity against HCC1954 than MCF7 cells, but weak toxicity against PBMC. This is the first report of the cytotoxic effect of A. muricata on HCC1954 cells, highlighting its potential for treating anti-estrogen-resistant breast cancers and low toxicity against PBMC.
Annona muricata Linn. (Annonaceae) es una planta tropical con múltiples efectos benéficos en la salud incluyendo propiedades antitumorales. En pacientes con cáncer de mama la sobreexpresión del oncogen HER2 corresponde a un mal pronóstico, por lo que el objetivo principal de este estudio fue evaluar la citotoxicidad de extractos etanólicos de hojas secas y frescas de A. muricata en células tumorales de mama HER2+. Se aplicaron pruebas de MTT y se determinaron IC50en células HCC1954 (HER2+); se utilizaron células MCF7 (HER-) y células mononucleares de sangre periférica (PBMC) como control. Se valoró también el contenido en polifenoles totales, y se realizó un tamizaje fitoquímico. El efecto citotóxico de los extractos de A. muricata (125-1000 µg/mL) fue dosis-dependiente y específico para cada tipo celular. Los extractos presentaron mayor actividad citotóxica contra HCC1954 en comparación con MCF7 y baja toxicidad contra PBMC. Este es el primer reporte del efecto citotóxico de A. muricata en HCC1954 y destaca su potencial terapéutico para tratamiento de cáncer de mama resistentes a antiestrógeno y baja citotoxicidad contra PBMC.
Assuntos
Neoplasias da Mama/prevenção & controle , Annona/química , Antineoplásicos/uso terapêutico , Plantas Medicinais , Neoplasias da Mama/tratamento farmacológico , Citotoxinas/farmacologiaRESUMO
A new series of 5-sustituted-3-((2-(4-nitrophenyl) Thiazol-4-yl) Imino)-1-(substituted-1-ylmethyl) Indolin-2-one derivatives (3a-3l) are synthesized by conventional method. All synthesized compounds were characterized via IR, 1H-NMR, 13C-NMR and MASS spectral analysis. Compounds were evaluated for their anticancer activity against MCF-7 cell lines and antibacterial activity against Staphylococcus aureus, Bacillus subtilis, E. coli and Salmonella paratyphi. These results indicated that compound 3a, 3e, 3f and 3j showed good activity compared to the standard drug. Synthesized target molecules exhibited anticancer activity against breast cancer cell lines with IC50 values of range from 60.21. to 130.21 ?g. Compound 3c (60.21?g) showed good activity compared with doxorubicin. Compounds were subjected to molecular docking studies with estrogen receptor (ER) epidermal growth factor receptor (EGFR) with PDB ID:3ERT. Among the docked ligands, compound 3a and 3e reported highest docking score (-8.512, -6.869) with glide binding energy (-43.785, -24.187).
RESUMO
Background: According to CDC, polycystic ovarian syndrome (PCOS) is responsible for infertility in women with 6-12% incidences all over the world. The current treatment options available have several side effects such as amenorrhea and obesity amongst others. Dietary interventions such as non-estrogenic and androgen-suppressing foods along with nutraceutical products is considered for treating PCOS with minimum side effects. One such product of Zenherbs lab called FertiZen-R™ is a combination of three phytochemicals from three plants viz., Foeniculum vulgare, Linum Usitatissimum, Glycyrrhiza glabra, hibiscus extract and is developed as a product to treat and control PCOS.Methods: The FertiZen-R™ was quantified for the presence of phytochemicals such as glycyrrhizic acid, polyphenols, and saponins. It was further tested for estrogenic/anti-estrogenic potential on estrogen-positive breast cancer cell line (MCF-7) using an E-Screen assay.Results: The FertiZen-R™ showed presence of 5% glycyrrhizinic acid, 5% saponins, and 2-3% polyphenols. It exhibited a strong anti-estrogenic potential with 40-50% inhibition from concentrations as low as 0.156 mg/ml like tamoxifen (IC50 at 0.156 mg/ml), while inositol, a natural growth promoter, showed no effect on the cell viability.Conclusions: FertiZen-R™ showed anti-estrogenic potential when tested in-vitro and can be used to treat PCOS in women even with ER-positive breast cancer cells. However, clinical studies to determine the dosage are required to warrant the potential of FertiZen-R™.
RESUMO
Breast cancer, the second most common cancer after lung cancer, is the most common cancer type diagnosed in women. No definitive treatment has been established for breast cancer yet, but essential fatty acids offer a promising option. Omega fatty acids are classified in the essential fatty acids that the body cannot produce and, therefore, must be taken through the foods of animal or plant origin. Although in the literature the omega fatty acids have been shown to exhibit significant positive effects in inhibiting various tumor types, their mechanism of action, the apoptotic pathways they employ, and the genes they control have not been clarified yet. In this study, various doses and combinations of omega-3 [Eicosapentaenoic acid (EPA), Docosahexaenoic acid (DHA)] and omega-6 [Linoleic acid (LA)] fatty acids were administered to human breast cancer MCF7 cell line for 24 h, and using the enzyme-linked immunosorbent assay (ELISA) method, the protein expression levels of the following apoptosis-related genes were determined: phospho-p53 (Ser15), p53, Bad, phospho-Bad (Ser112), cleaved Caspase-3 (Asp175), and cleaved PARP (Asp214). Even though there was no significant difference observed in the expressions of phospho-p53 (Ser15) and p53 at all doses, other protein expressions were found to increase significantly, suggesting that Omega-3 and -6 can mediate apoptotic pathway to induce cell death in breast cancer cells.
RESUMO
Objective:To investigate the effect of miR-27b-3p on radiation resistance of breast cancer cells.Methods:The relative expression levels of miR-27b-3p in normal tissues and breast cancer tissues were analyzed through GEO database and verified by the qRT-PCR assay. Cloning formation, immunofluorescence, and EDU assay were used to assess the functions of miR-27b-3p on radioresistance of breast cancer cells. The luciferase reporter assay was used to verify whether miR-27b-3p directly targeted PLK2 mRNA. A rescue experiment was performed to identify the influence of PLK2 overexpression on miR-27b-3p regulated radioresistance.Results:The expression level of miR-27b-3p in both breast cancer tissues ( t=2.99, P<0.01) and breast cancer cells ( t=21.21, 32.88, P<0.05) was significantly higher than those in normal breast tissues and cells, especially, it was elevated in radioresistant MCF-7R cells ( t=25.63, P<0.05). Overexpression of miR-27b-3p enhanced the cloning efficiency of MCF-7 cells ( t=10.32, P<0.05), and had a protective effect on the proliferation of irradiated MCF-7 cells ( t=8.77, 8.26, 8.03, P<0.05). But interference of miR-27b-3p reduced the cloning efficiency and proliferation of MCF-7R cells ( t=40.00, P<0.05) after irradiation with different doses( t=8.54, 8.32, 8.23, P<0.05). Moreover, PLK2 was verified to be a direct target of miR-27b-3p, and overexpression of PLK2 inhibited miR-27b-3p-mediated radioresistance of breast cancer cells (MCF-7: t=9.66, P<0.05; MCF-7R: t=6.42, P<0.05). Conclusions:miR-27b-3p contributes to the radioresistance of breast cancer cells by targeting PLK2.
RESUMO
@#[摘 要] 目的:探讨1, 25-二羟维生素D3(VD3)对人乳腺癌MCF-7细胞凋亡的影响及其作用机制。方法:取体外培养的人乳腺癌MCF-7细胞,随机分为6组:对照组、2-脱氧葡萄糖(2-DG,葡萄糖抑制剂)组、1 µmol/L VD3组、10 µmol/L VD3组、2-DG+1 µmol/L VD3组和2-DG+10 µmol/L VD3组。药物干预6组细胞48 h后,以葡萄糖摄取测定试剂盒检测细胞的葡萄糖摄取量、ATP试剂盒检测细胞中ATP含量和乳酸试剂盒检测细胞的乳酸水平,WB法检测MCF-7细胞中细胞色素C(Cyt c)和凋亡相关蛋白(Bcl-2、BAX、PARP1、caspase9和caspase3)的表达水平。结果:与对照组比较,VD3干预后,MCF-7细胞的凋亡率明显增加(P<0.05或P<0.01),同时细胞的葡萄糖摄取量、ATP含量及乳酸水平均明显降低(P<0.05或P<0.01),Cyt c、BAX、PARP1、caspase9及caspase3蛋白表达量明显升高(均P<0.05),Bcl-2蛋白表达量降低(P<0.05或P<0.01);VD3联合2-DG干预后,各组细胞检测指标的变化更为明显(P<0.05或P<0.01)。结论:VD3可通过抑制人乳腺癌MCF-7细胞的糖酵解过程并以线粒体的Cyt c途径促进细胞凋亡。
RESUMO
Aim To investigate the radiosensitizing effect of the PARP inhibitor Olaparib on MCF-7 breast cancer model and to monitor the radiosensitizing effect of Olaparib by F-Fluoroerythronitroimidazole (F-FETNIM) PET/CT. Methods MCF-7 breast cancer model was established and divided into control group, Olaparib group, irradiation group and Olaparib + irradiation group according to random number table method; tumor volume was measured to calculate tumor inhibition rate and survival time of tumor-bearing mice was counted.
RESUMO
@#[摘 要] 目的:探讨中药地黄提取物梓醇(Cat)对乳腺癌MCF-7细胞增殖与凋亡,以及裸鼠移植瘤生长的影响及其机制。方法:以不同质量浓度(0、5、25、50、100、200 μg/mL)Cat处理人乳腺癌MCF-7细胞,用MTT法筛选Cat给药浓度。将MCF-7细胞分为空白对照组、Cat低剂量组、Cat中剂量组、Cat高剂量组、Cat+sh-NC组和Cat+sh-FOXO3组,采用Edu细胞增殖实验、平板克隆实验、流式细胞术分别检测各组细胞的增殖与克隆形成能力、凋亡率和细胞周期,WB法检测各组细胞中FOXO3、FOXM1、caspase-3和caspase-8蛋白表达。构建乳腺癌MCF-7细胞裸鼠移植瘤模型,观察Cat对移植瘤生长的影响,WB法检测移植瘤组织中FOXO3和FOXM1蛋白表达。结果:Cat低(50 μg/mL)、中(100 μg/mL)、高(200 μg/mL)剂量处理的MCF-7细胞的增殖能力均显著下降(均P<0.05)。与空白对照组比较,Cat低、中、高剂量组Edu阳性细胞率、克隆形成数、S期与G2/M期细胞比例及FOXO3蛋白表达均显著降低(均P<0.05),细胞凋亡率、G0/G1期细胞比例及FOXM1、caspase-3、caspase-8蛋白表达均显著升高(均P<0.05);与Cat+sh-NC组比较,Cat+sh-FOXO3组Edu阳性细胞率、克隆形成数、S期与G2/M期细胞比例及FOXO3蛋白表达均显著升高(均P<0.05),细胞凋亡率、G0/G1期细胞比例及FOXM1、caspase-3和caspase-8蛋白表达均显著下降(均P<0.05)。Cat组MCF-7细胞裸鼠移植瘤体积、质量和FOXO3蛋白表达均显著降低(均P<0.05),FOXM1的蛋白表达显著升高(P<0.05)。结论:Cat抑制乳腺癌MCF-7细胞增殖并促进凋亡,在体内抑制裸鼠移植瘤的生长,其机制可能与上调FOXO3、下调FOXM1的表达有关。
RESUMO
Introduction@#Breast cancer is the most common cancer among women in the Philippines and about 3 in every 100 Filipina will be diagnosed with breast cancer in their lifetime. There is a need to discover safe, yet inexpensive herbal extracts with potential cytotoxic properties as potential treatment modalities to treat breast cancer. @*Objectives@#This study seeks to explore the cytotoxic and apoptotic properties of the ethyl acetate fraction of the defatted crude methanol leaf extract of Syzygium samarangense in MCF-7 breast cancer cell lines. @*Methods@#Screening for flavonoids of the extracts was performed using TLC, total flavonoids, total phenols, FTIR and LC-MS spectroscopy. The hydrogen peroxide and ferric reducing anti-oxidant power were used as substrates to assess in vitro anti-oxidative properties of the extracts. The MTT dye viability assay was used to assess the cytotoxic properties of the extracts against MCF-7 cells. Apoptotic properties of the extracts in MCF-7 cells were determined by caspase-3 activation assay, DNA fragmentation patterns and fluorescence microscopy after annexin-V and propidium iodide staining. @*Results@#The abundance of flavonoids in the ethyl acetate fraction of the crude methanol leaf extract was established by TLC, FTIR, LC-MS/MS, total flavonoid and total phenol analyses. The in vitro anti-oxidative properties of this extract was comparable to ascorbic acid. The median inhibitory concentration (IC50) of this extract in MCF-7 breast cancer cell lines was 7.2 mcg/mL while doxorubicin registered an IC50 of 1.2 mcg/mL. At this concentration, the extract was not cytotoxic to normally-dividing breast epithelial cells. Cytotoxicity of the extract was mediated via apoptosis as demonstrated by DNA fragmentation, caspase-3 activation and fluorescence microscopic analyses. @*Conclusion@#The study shows that the flavonoid-rich ethyl acetate fraction of the crude methanol leaf extract of S. samarangense possesses potent apoptotic and cytotoxic properties against MCF-7 breast cancer cell lines at low concentrations.
Assuntos
Células MCF-7 , SyzygiumRESUMO
@#Introduction: Aquilaria malaccensis, also known as “Pokok Karas” in Malaysia, is widely used in Southeast Asian countries for the treatment of joint pain, diarrhoea and inflammatory diseases, and has shown beneficial effects as an anticancer agent. The aim of this study was to investigate the effect of ethanol leaf extracts of A. malaccensis on MCF-7 cells. Methods: MTT-based cytotoxic and antiproliferative assay was used to determine the outcome of ethanolic extract toward MCF-7 cells. The mode of cell death was determined by the AO/PI double staining assay and the depolarisation of the mitochondria membrane potential. Results: IC50 value of the extract against MCF-7 cells treated for 72 hours was 4.1 ± 2.08 µg/mL, while the IC50 value for doxorubicin was 2.92 ± 0.12 µg/mL. The extract showed a lower cytotoxic effect against the NIH/3T3 cells and inhibited the growth of MCF-7 cells in a dose dependent manner. AO/PI double stain showed that the ethanolic extract of A. malaccensis leaves induced MCF-7 cells into apoptotic cell death. The present study showed that the ethanolic extract of A. malaccensis induced apoptosis through mitochondrial pathway as indicated by its ability to take up JC-1. Conclusion: The study found that ethanolic extract obtained from A. malaccensis leaves is cytotoxic on MCF-7 cells, resulting to apoptotic cell death of the cells.
RESUMO
This study aimed to evaluate in vitro the possible mechanisms underlying the estrogenic potential of benzalkonium chloride (BAC) as a disinfectant emerging contaminant. Effects of BAC at the environmentally-relevant concentrations on estrogen synthesis and estrogen receptor (ER) signaling were assessed using the H295R steroidogenesis assay and the MCF-7 proliferation assay, respectively. Results showed that exposure to BAC at concentrations of 1.0-1.5 mg/L for 48 h significantly increased estradiol production of H295R cells in a concentration-dependent manner. Transcription of steroidogenic genes 3β‐HSD2, 17β‐HSD1, 17β‐HSD4, and CYP19A were significantly enhanced by BAC. In ER-positive MCF-7 cells, exposure to 0.5-1.5 mg/L BAC for 48 h significantly promoted cell proliferation and increased the expressions of ERα and G-protein coupled estrogen receptor 1. Flow cytometry analysis showed that 0.5-1.5 mg/L BAC significantly decreased the percentage of cells in G0/G1 phase, increased the percentage in S phase, and BAC at concentrations of 1.0 and 1.5 mg/L increased the G2/M phase cells. Findings of the study suggested that BAC at environmentally-relevant concentrations might act as a xenoestrogen through its inhibitory effect on steroidogenesis and ER-mediated mechanism.
RESUMO
SUMMARY OBJECTIVE: We aimed to examine the potential anticancer effects of ozone applied after chemotherapeutic treatment with different concentrations of doxorubicin in Luminal-A subtype of human breast cancer cell line (MCF-7) and compare the results with effects on L929 fibroblast cell line. METHODS: Both cell lines were incubated with increasing doses of doxorubicin (1-50 μM) for 24 h at 37°C. Then, half of groups were incubated with 30 μg/mL ozone for 25 min as combination groups. Cell viability was analyzed by MTT assay, apoptosis by flow cytometry, and levels of tumor necrosis factor alpha, transforming growth factor beta, and matrix metalloproteinase-2 and MMP-9 by immunocytochemistry. RESULTS: Doxorubicin + ozone treatment enhanced viability of L929 (p<0.01) but reduced viability of MCF-7 compared to only doxorubicin-applied cells without ozone treatment (p<0.001). This combined treatment also enhanced apoptotic effect of doxorubicin on MCF-cells (p<0.001), but not on L929. It significantly increased all protein levels of L929 compared with those of other groups (p<0.05 for tumor necrosis factor alpha and MMP-2; p<0.01 for transforming growth factor beta and MMP-9). This treatment reversed the effect of doxorubicin on tumor necrosis factor alpha levels and considerably reduced MMP-2 and MMP-9 levels of MCF-7 compared with those of control group (p<0.01 and p<0.001, respectively). CONCLUSION: Ozone treatment potentiated the apoptotic and anticancer activities of doxorubicin in MCF-7 cells and showed repairing and healing effect on healthy fibroblast cells, which were damaged from cytotoxic effects of chemotherapeutic agent. MCF-7 cells may acquire sensitivity against the doxorubicin combined with ozone treatment through activating tumor necrosis factor alpha, MMP-2, and MMP-9 expressions.
RESUMO
SUMMARY OBJECTIVE: The phosphoinositide 3-kinase/protein kinase AKT/mammalian target of rapamycin signaling pathway is essential for proper cellular metabolism and cell growth. However, aberrant activation of this pathway has been linked to the progression and metastasis of breast cancer. Recently, the role of long non-coding RNAs in interfering with the cell signaling pathways involved in cell growth and metabolism has been identified. HOX antisense intergenic RNA is an long non-coding RNA whose abnormal expression has been associated with development, therapy resistance, and metastasis of breast cancer. The purpose of this study was to investigate whether the long non-coding RNA HOX antisense intergenic RNA is linked to the phosphoinositide 3-kinase/protein kinase AKT/mammalian target of rapamycin signaling pathway in breast cancer cells. METHODS: HOX antisense intergenic RNA was silenced in the breast cancer cell line MCF-7 using siRNAs. Subsequently, the gene expression level of HOX antisense intergenic RNA, PI3K, AKT, and mTOR was assessed using real-time RT-PCR. Also, the 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyl-tetrazolium bromide) assay was used to analyze cell proliferation. RESULTS: The results revealed that HOX antisense intergenic RNA knockdown can downregulate the expression of PI3K, AKT, and mTOR RNAs compared to negative control in MCF-7 cells. In addition, the proliferation of breast cancer cells was significantly reduced following the HOX antisense intergenic RNA silencing. CONCLUSION: This study may introduce HOX antisense intergenic RNA as a molecule involved in the upregulation of the phosphoinositide 3-kinase/protein kinase AKT/mammalian target of rapamycin signaling pathway in breast cancer cells that may contribute to breast cancer cell proliferation.
RESUMO
Introducción: En el cáncer de mama no se ha reportado una bacteria como causante, pero sí la presencia de diferentes especies que se relacionan de forma beneficiosa o perjudicial. Objetivo: Identificar la microbiota presente en nódulos mamarios y sus efectos morfológicos sobre la línea celular MCF-7. Métodos: Estudio exploratorio experimental de una población de 57 mujeres con nódulos mamarios, intervenidas para biopsia por punción en un centro de diagnóstico. De estas, se escogieron 17 muestras mediante muestreo no probabilístico para el análisis metagenómico. Se realizó una infección con células MCF-7 y Staphylococcus saprophyticus a MOI de 1:1, 5:1 y 10:1 (48 horas de exposición). Resultados: De las 57 muestras tomadas, solo en 7 pacientes se obtuvo resultado positivo (12,28 por ciento) y en el resto (50) no hubo crecimiento bacteriano. Por metagenómica se obtuvo la microbiota siguiente: Proteobacteria (47 por ciento), Escherichia (9,4 por ciento) y Yokenella (8,2 por ciento), entre otros. Los controles fueron negativos. Solo dos pacientes resultaron positivas para cáncer, entre ellas la especie común fue S. saprophyticus. En la infección, los cambios morfológicos se evidenciaron desde la MOI 5:1. Conclusión: El bacterioma extraído de los nódulos de una población femenina es en su mayoría flora endógena del órgano mamario(AU)
Introduction: No bacterium has been reported as causative of breast cancer, but the presence of different species that are related in a beneficial or detrimental way. Objective: To identify the microbiota present in breast nodules and its morphological effects on the MCF-7 cell line. Methods: Exploratory, experimental study of a population of 57 women with breast nodules, operated for needle biopsy in a diagnostic center. Of these, 17 samples were chosen by non-probabilistic sampling for metagenomic analysis. Infection was performed with MCF-7 cells and Staphylococcus saprophyticus at MOI of 1:1, 5:1 and 10:1 (48 hours of exposure). Results: Of the 57 samples taken, only 7 yielded positive results (12.28 percent) and the rest (50) had no bacterial growth. The following microbiota was obtained by metagenomics: Proteobacteria (47 percent), Escherichia (9.4 percent) and Yokenella (8.2 percent), among others. Controls were negative. Only two patients tested positive for cancer, and S. saprophyticus was the common species. In the infection, morphological changes were evidenced from MOI 5:1. Conclusion: The bacteriome extracted from the nodules of a female population is mostly endogenous flora of the mammary glands(AU)
Assuntos
Humanos , FemininoRESUMO
In breast cancer treatment, chemotherapy resistance is a major problem where many receptive tumors rebound and develop resistance. When provided in combination, cancer drugs are most successful, thus reducing the risk of developing resistant cancer cells. However, the evaluation of combination therapies has increased rapidly in recent years. Consequently, by repurposing old treatments, the discovery of additional medicines that may interact synergistically with chemotherapy is considered a current medical aim through discovering a new cancer medication or therapeutic strategy. The purpose of this research is to increase the anti-cancer activity of carboplatin (CP) by increasing the apoptotic effect of breast cancer cells (MCF-7) during in vitro experiments in combination with oxytetracycline. Our results showed a high synergistic effect between oxytetracycline and carboplatin, MCF-7 representative cell treated with carboplatin with/without different concentrations of oxytetracycline (5% and 10% of IC50). Oxytetracycline, which potentiated the action of carboplatin and/or had notable activity was reported as a single agent. This research demonstrated the synergistic relationship between oxytetracycline and carboplatin in viability assays. Surprisingly, our findings suggest that inhibiting treatment strategies can extend carboplatin's therapeutic window, potentially allowing for cancer therapy.(AU)
No tratamento do câncer de pulmão a resistência à quimioterapia é o maior problema no qual muitos tumores receptivos apresentam um rebote e desenvolvem a resistência. Quando oferecidas em combinações, as drogas anticancerígenas apresentam maior taxa de sucesso, reduzindo assim o risco de desenvolvimento de células cancerígenas resistentes. Contudo, a avaliação das terapias de combinação tem crescido muito rapidamente. Consequentemente, por reaproveitamento de tratamentos antigos, a descoberta de novos medicamentos adicionais que podem interagir sinergicamente com a quimioterapia que é considerada como auxílio médico na corrente busca à descoberta de novas medicações anticancerígenas ou estratégias terapêuticas. O propósito da presente pesquisa é aumentar a atividade anticancerígena da Carboplatina (CP) pelo incremento do efeito apoptótico de células de câncer pulmonar (MCF-7) em experimentos in vitro pela combinação com oxitetraciclina. Os resultados obtidos confirmaram elevado efeito sinérgico entre oxitetraciclina e Carboplatina em células MCF-7 representativas tratadas com Carboplatina, com e sem diferentes concentrações de oxitetraciclina (5% e 10% de IC50). A oxitetracilina que potencializou a ação da Carboplatina e/ou teve uma notável atividade relatada como um agente isolado. A pesquisa demonstrou a relação sinérgica entre oxitetraxiclina e Carboplatina nos ensaios de viabilidade. Surpreendentemente, os resultados obtidos sugeriram que as estratégias de tratamento inibidor podem aplicar um janela terapêutica da Carboplatina com potencial para a terapia do câncer.(AU)
Assuntos
Oxitetraciclina/farmacologia , Neoplasias da Mama/tratamento farmacológico , Carboplatina/farmacologia , Técnicas In Vitro/veterinária , Sinergismo Farmacológico , Células MCF-7/efeitos dos fármacosRESUMO
OBJECTIVE To study in vitro inhibitory effects of realgar nanoparticles on breast cancer stem cells. METHODS Human breast cancer MCF- 7 parent cells were selected as subjects and cultured by serum-free culture to obtain breast cancer stem cells. Using adriamycin (1 mg/L)as positive control ,same concentration of water-processed realgar as reference ,the effects of realgar nanoparticles on the proliferation of MCF- 7 parent cells and stem cells were detected by CCK- 8 method. The effects of realgar nanoparticles on the formation of mammosphere ,the ability of differentiation ,migration and invasion ,the proportion of CD44+/CD24- subgroup in breast cancer stem cells were detected by mammosphere formation and differentiation experiment , scratch experiment ,Transwell invasion experiment and flow cytometry. Western blot assay was used to detect the expression of proteins related to epithelial mesenchymal transformation pathway (E-cadherin and vimentin ) in breast cancer stem cells. RESULTS The survival rates of MCF- 7 parent cells and stem cells (except for breast cancer stem cells in both 1 mg/mL groups )in 1,5,10,40,60,80 mg/L groups of water-processed realgar and realgar nanoparticles were significantly lower than blank control group(P<0.01). The number of mammosphere (>20 stem cells )in 1,2.5,5,10 mg/L groups of water-processed realgar and realgar nanoparticles was significantly lower than blank control group (P<0.01);the volume of mammosphere decreased and the differentiated adherent cells decreased ;the healing rate of wound ,relative invasion rate (except for water-processed realgar 1 mg/L group)and the proportion of CD 44+/CD24- subgroup were significantly lower than blank control group (P<0.01). The expressions of E-cadherin in 2.5,10 mg/L groups of water-processed realgar and realgar nanoparticles was significantly higher than blank control group ,and the expressions of vimentin was significantly lower than those in blank control group (P<0.01). The above effects of realgar nanoparticles were generally better than those of water-processed realgar with the same mass concentration (P< 0.01). CONCLUSIONS Compared with water-processed realgar with the same mass concentration ,realgar nanoparti cles can significantly inhibit the proliferation of breast cancer stem cells, the formulation and differential ability of mammo- sphere,and reduce the proportion of CD 44+/CD24- subgroup. The effect may be associated with the inhibition of migration and invasion of breast cancer stem cells by inhibiting the expression of proteins related to epithelial mesenchymal transformation pathway.
RESUMO
@#[摘 要] 目的:探讨干扰B7-H4表达对乳腺癌细胞增殖、凋亡、周期以及相关下游分子表达的影响。方法:利用脂质体转染技术分别将特异性靶向B7-H4的siRNA(siB7-H4)及其阴性对照(siNC)转染至对数生长期的乳腺癌T47D和MCF-7细胞,分别命名为T47D-siB7-H4、T47D-siNC、MCF-7-siB7-H4和MCF-7-siNC组。用qPCR法和WB法验证siRNA干扰效果及其对细胞周期分子cyclin D1表达的影响,CCK-8法和FCM分别检测干扰B7-H4表达对T47D和MCF-7细胞增殖、周期和凋亡的影响,qPCR法检测B7-H4干扰对E2F家族相关转录因子表达的影响。结果:成功构建干扰B7-H4表达的乳腺癌T47D和MCF-7细胞。与T47D-siNC和MCF-7-siNC组相比,T47D-siB7-H4和MCF-7-siB7-H4组细胞中B7-H4 mRNA和蛋白表达水平均显著降低、细胞增殖能力显著降低(均P<0.01),并伴有G1/S期细胞周期阻滞以及cyclin D1表达下调(均P<0.01),但细胞凋亡率差异无统计学意义(均P>0.05)。与T47D-siNC相比,干扰B7-H4后T47D细胞中E2F1、E2F2、E2F7和E2F8 mRNA水平有不同程度的降低(均P<0.01);与MCF-7-siNC相比,干扰B7-H4后MCF-7细胞中E2F1、E2F2、E2F3、E2F7和E2F8 mRNA水平均有不同程度的降低(P<0.05或P<0.01)。结论:干扰乳腺癌细胞B7-H4表达可下调cyclin D1和E2F家族相关转录因子的表达,导致细胞周期阻滞并抑制细胞增殖。
RESUMO
@#[摘 要] 目的:探讨miR-620对乳腺癌MCF-7细胞放射敏感性的影响及其机制。方法:收集2017年3月至2018年3月在海南省儋州市人民医院手术切除的21例乳腺癌患者的癌及癌旁组织标本,以及乳腺癌细胞MCF-7、BCaP-37和乳腺上皮细胞HBL-100,采用qPCR法检测癌组织和细胞中miR-620和生长抑制因子4(ING4)mRNA的表达。利用脂质体转染技术,分别将miR-620抑制剂(anti-miR-620)和抑制剂阴性对照(anti-miR-NC)、anti-miR-620和ING4小干扰RNA(si-ING4)、anti-miR-620和小干扰RNA阴性对照序列(si-NC)转染至MCF-7细胞,经放射处理后(依次记为IR+anti-miR-620组、IR+anti-miR-NC组、IR+anti-miR-620+si-ING4组、IR+anti-miR-620+si-NC组),利用克隆形成实验、MTT法和FCM分别检测细胞放射敏感性、细胞增殖活力、细胞周期分布和凋亡率。双荧光素酶报告基因实验和WB法验证miR-620和ING4的靶向关系。结果:与癌旁组织和HBL-100细胞比较,乳腺癌组织和细胞中miR-620表达均显著升高(均P<0.01)、ING4 mRNA表达均显著降低(均P<0.01)。与IR+anti-miR-NC组比较,IR+anti-miR-620组MCF-7细胞增殖活力、S期细胞比例均显著降低(均P<0.01),细胞凋亡率、G0-G1期细胞比例、放射敏感性均显著升高(均P<0.01)。与IR+anti-miR-620+si-NC组比较,IR+anti-miR-620+si-ING4组MCF-7细胞增殖活力、S期细胞比例均显著升高(均P<0.01),细胞凋亡率、G0-G1期细胞比例和放射敏感性均显著降低(均P<0.01)。双荧光素酶报告基因实验证明ING4是miR-620的靶基因,miR-620靶向负性调控ING4表达。结论:敲减miR-620可能通过上调ING4表达抑制乳腺癌MCF-7细胞增殖,并促进细胞凋亡和放射敏感性。