Effects of Rhizopus Nigricans Exopolysaccharide on Proliferation, Apoptosis, and Migration of Breast Cancer MCF-7 Cells and Akt Signaling Pathway
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Abstract:
Objective. To study the effect of Rhizopus nigricans exopolysaccharide EPS1-1 on the proliferation, apoptosis, and migration of breast cancer MCF-7 cells. Methods. Human breast cancer MCF-7 cells were cultured in vitro and treated with different concentrations of EPS1-1. The effect of EPS1-1 on cell proliferation was tested by the CCK-8 experiment, and the effect of EPS1-1 on cell apoptosis was determined by flow cytometry. And the scratch test was used to detect the impact of EPS1-1 on cell migration. Western blot then was used to measure the expression changes of related proteins in the Akt signaling pathway. Results. Compared with the control group, treatment with EPS1-1 significantly reduced the proliferation, migration, and invasion ability of MCF-7 cells and promoted the apoptosis of MCF-7 cells in a dose-dependent manner. In terms of the underlying mechanism, EPS1-1 can significantly inhibit the phosphorylation of Akt at threonine 308 and serine 473 and cause the expression changes of downstream proliferation-related genes CCND1 and p21, apoptosis-related genes Bcl-2 and Bax, and migration-related genes Vimentin and E-cadherin in terms of their protein levels. Conclusion. EPS1-1 can inhibit the proliferation, migration, and invasion of breast cancer MCF-7 cells and promote the apoptosis of MCF-7 cells by inhibiting the activation of the Akt signaling pathway. Therefore, EPS1-1 can be used as a potential new drug or adjuvant drug for the treatment of breast cancer.Keywords:
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Background and Objectives: Angiogenesis is the process of forming new capillary vessels from existing ones.This process is implicated in tumour growth, metastasis and wound healing.During tumour metastasis, angiogenesis is amplified as new capillaries are needed for cancer to spread.However, when this process is inhibited, wound healing is impaired.Cyclooxygenase-2 (COX-2), nitric oxide synthase (NOS) and lipoxygenase (LOX) are some of the enzymes that are highly expressed during angiogenesis and therefore provide a useful way to detect angiogenesis.Some medicinal plants have shown to possess angiogenetic-modulating properties, which has led to the development of anti-angiogenic drugs for the treatment of cancer.Spirostachys africana sond has been used historically to treat open wounds, ulcers and cancer.Nevertheless, its mechanism of action is still unclear.The aim of this study was to determine mechanisms of anticancer and wound healing activities of S. africana by evaluating its effects on the proliferation of MCF-7 cells and on the activities of COX-2, LOX and NOS.Materials and Methods: The dried plant materials were extracted with water and sequentially with organic solvents in their order of increasing polarity.Extracts were screened for cell growth inhibitory activity against breast cancer MCF-7 cells and for selectivity against normal breast MCF-10A cells.Extracts that showed growth inhibitory activity with IC 50 values <10 µg mLG 1 were further evaluated for effects on COX-2, LOX and NOS enzymatic activity.Results: The non-polar extracts of all plant parts had anti-proliferation activity with IC 50 of 10 µg mLG 1 or lower.All the leaf extracts showed selectivity for MCF-7 breast cancer.The selected extracts also induced NOS activity and inhibited LOX and COX-2 activity in a concentration dependent manner.Conclusion: It was concluded that the selected extracts may suppress angiogenesis by inhibiting COX-2 and LOX and induce apoptosis by increasing NOS activity.Spirostachys africana was found to contain tannins, glycosides, saponins and alkaloids which could be responsible for the biological activity observed.
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This study analyzes the uptake and antiproliferative effect of two different chemical forms of iodine, iodide (I − ) and molecular iodine (I 2 ), in MCF-7 cells, which are inducible for the Na + /I − symporter (NIS) and positive for pendrin (PDS). The mouse fibroblast cell line NIH3T3 was used as control. Our results show that in MCF-7 cells, I − uptake is sustained and dependent on NIS, whereas I 2 uptake is transient with a maximal peak at 10 min and a final retention of 10% of total uptake. In contrast, no I − was taken up by NIH3T3 cells, and although I 2 was captured with the same time pattern as in MCF-7 cells, its uptake was significantly lower, and it was not retained within the cell. The uptake of I 2 is independent of NIS, PDS, Na + , and energy, but it is saturable and dependent on protein synthesis, suggesting a facilitated diffusion system. Radioiodine was incorporated into protein and lipid fractions only with I 2 treatment. The administration of non-radiolabeled I 2 and 6-iodo-5-hydroxy-8,11,14-eicosatrienoic acid (6-iodolactone, an iodinated arachidonic acid), but not KI, significantly inhibited proliferation of MCF-7 cells. Proliferation of NIH3T3 cells was not inhibited by 20 μM I 2 . In conclusion, these results demonstrate that I 2 uptake does not depend on NIS or PDS; they suggest that in mammary cancer cells, I 2 is taken up by a facilitated diffusion system and then covalently bound to lipids or proteins that, in turn, inhibit proliferation.
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Cancer is one of the leading causes of death in the world. Breast cancer is the most common form of cancer among women and is responsible for 15 % of all cancer related deaths. Though there are significant advancements in cancer treatment strategies, clinical tumour treatment methods currently employed are often accompanied by severe side effects as they induce damage to the normal cells along with the cancer cells. The alterations in the biophysical and biomechanical properties of a cell as it undergoes transformation from a normal to cancerous cell results in changes in its dynamic characteristics. These changes can be utilised to induce selective cytotoxicity of tumour cells. In the present study, two simulation models (homogenous and non-homogenous) of normal (MCF-10A) and cancerous (MCF-7) breast cells are developed. A finite element approach using Ansys is adopted to investigate the variation in dynamic characteristics of the cells using the two modelling approaches. Results indicate that the natural frequencies of cells modelled as a homogenous system is greater than that of cells whose sub-cellular material properties are considered for analysis. A comparison of the first four natural frequencies using the two modelling approaches for both MCF-10A and MCF-7 cells are illustrated and the corresponding mode shapes reported. A plot highlighting the variation in the natural frequencies of MCF-10A and MCF-7 using the two modelling approaches is presented.
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Multidrug resistance is a serious problem in modern medicine and the reason for the failure of various therapies. A particularly important problem is the occurrence of multidrug resistance in cancer therapies which affects many cancer patients. Observations on the effect of metformin-a well-known hypoglycemic drug used in the treatment of type 2 diabetes-on cancer cells indicate the possibility of an interaction of this substance with drugs already used and, as a result, an increase in the sensitivity of cancer cells to cytostatics. The aim of this study was to evaluate the effect of metformin on the occurrence of multidrug resistance of breast cancer cells. The MCF-7-sensitive cell line and the MCF-7/DX cytostatic-resistant cell line were used for this study. WST-1 and LDH assays were used to evaluate the effects of metformin and doxorubicin on cell proliferation and viability. The effect of metformin on increasing the sensitivity of MCF-7 and MCF-7/DX cells to doxorubicin was evaluated in an MDR test. The participation of metformin in increasing the sensitivity of resistant cells to the effect of the cytostatic (doxorubicin) has been demonstrated.
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