{"title":"人羊水间充质干细胞条件培养基对人MCF-7乳腺癌细胞系凋亡的影响","authors":"Roghiyeh Pashaei-Asl, Maryam Pashaiasl, Esmaeil Ebrahimie, Maryam Lale Ataei, Maliheh Paknejad","doi":"10.34172/bi.2022.23813","DOIUrl":null,"url":null,"abstract":"<p><p></p><p><strong>Introduction: </strong>Breast cancer, as the most common malignancy among women, is shown to have a high mortality rate and resistance to chemotherapy. Research has shown the possible inhibitory role of Mesenchymal stem cells in curing cancer. Thus, the present work used human amniotic fluid mesenchymal stem cell-conditioned medium (hAFMSCs-CM) as an apoptotic reagent on the human MCF-7 breast cancer cell line.</p><p><strong>Methods: </strong>Conditioned medium (CM) was prepared from hAFMSCs. After treating MCF-7 cells with CM, a number of analytical procedures (MTT, real-time PCR, western blot, and flow cytometry) were recruited to evaluate the cell viability, Bax and Bcl-2 gene expression, P53 protein expression, and apoptosis, respectively. Human fibroblast cells (Hu02) were used as the negative control. In addition, an integrated approach to meta-analysis was performed.</p><p><strong>Results: </strong>The MCF-7 cells' viability was decreased significantly after 24 hours (<i>P</i> < 0.0001) and 72 hours (<i>P</i> < 0.05) of treatment. Compared with the control cells, Bax gene's mRNA expression increased and Bcl-2's mRNA expression decreased considerably after treating for 24 hours with 80% hAFMSCs-CM (<i>P</i> = 0.0012, <i>P</i> < 0.0001, respectively); an increasing pattern in P53 protein expression could also be observed. The flow cytometry analysis indicated apoptosis. Results from literature mining and the integrated meta-analysis showed that hAFMSCs-CM is able to activate a molecular network where Bcl2 downregulation stands in harmony with the upregulation of P53, EIF5A, DDB2, and Bax, leading to the activation of apoptosis.</p><p><strong>Conclusion: </strong>Our finding demonstrated that hAFMSCs-CM presents apoptotic effect on MCF-7 cells; therefore, the application of hAFMSCs-CM, as a therapeutic reagent, can suppress breast cancer cells' viabilities and induce apoptosis.</p>","PeriodicalId":48614,"journal":{"name":"Bioimpacts","volume":null,"pages":null},"PeriodicalIF":2.2000,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/0b/4b/bi-13-191.PMC10329748.pdf","citationCount":"1","resultStr":"{\"title\":\"Apoptotic effects of human amniotic fluid mesenchymal stem cells conditioned medium on human MCF-7 breast cancer cell line.\",\"authors\":\"Roghiyeh Pashaei-Asl, Maryam Pashaiasl, Esmaeil Ebrahimie, Maryam Lale Ataei, Maliheh Paknejad\",\"doi\":\"10.34172/bi.2022.23813\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p></p><p><strong>Introduction: </strong>Breast cancer, as the most common malignancy among women, is shown to have a high mortality rate and resistance to chemotherapy. Research has shown the possible inhibitory role of Mesenchymal stem cells in curing cancer. Thus, the present work used human amniotic fluid mesenchymal stem cell-conditioned medium (hAFMSCs-CM) as an apoptotic reagent on the human MCF-7 breast cancer cell line.</p><p><strong>Methods: </strong>Conditioned medium (CM) was prepared from hAFMSCs. After treating MCF-7 cells with CM, a number of analytical procedures (MTT, real-time PCR, western blot, and flow cytometry) were recruited to evaluate the cell viability, Bax and Bcl-2 gene expression, P53 protein expression, and apoptosis, respectively. Human fibroblast cells (Hu02) were used as the negative control. In addition, an integrated approach to meta-analysis was performed.</p><p><strong>Results: </strong>The MCF-7 cells' viability was decreased significantly after 24 hours (<i>P</i> < 0.0001) and 72 hours (<i>P</i> < 0.05) of treatment. Compared with the control cells, Bax gene's mRNA expression increased and Bcl-2's mRNA expression decreased considerably after treating for 24 hours with 80% hAFMSCs-CM (<i>P</i> = 0.0012, <i>P</i> < 0.0001, respectively); an increasing pattern in P53 protein expression could also be observed. The flow cytometry analysis indicated apoptosis. 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引用次数: 1
摘要
导读:乳腺癌是妇女中最常见的恶性肿瘤,具有高死亡率和化疗耐药的特点。研究表明间充质干细胞在治疗癌症方面可能具有抑制作用。因此,本研究使用人羊水间充质干细胞条件培养基(hAFMSCs-CM)作为人MCF-7乳腺癌细胞系的凋亡试剂。方法:从hAFMSCs制备条件培养基(CM)。用CM处理MCF-7细胞后,采用MTT、real-time PCR、western blot和流式细胞术等多种分析方法分别评估细胞活力、Bax和Bcl-2基因表达、P53蛋白表达和凋亡情况。以人成纤维细胞(Hu02)为阴性对照。此外,采用综合方法进行meta分析。结果:24h后MCF-7细胞活力明显降低(P P P = 0.0012, P)结论:hAFMSCs-CM对MCF-7细胞有凋亡作用;因此,应用hAFMSCs-CM作为治疗试剂,可以抑制乳腺癌细胞的生存能力,诱导细胞凋亡。
Apoptotic effects of human amniotic fluid mesenchymal stem cells conditioned medium on human MCF-7 breast cancer cell line.
Introduction: Breast cancer, as the most common malignancy among women, is shown to have a high mortality rate and resistance to chemotherapy. Research has shown the possible inhibitory role of Mesenchymal stem cells in curing cancer. Thus, the present work used human amniotic fluid mesenchymal stem cell-conditioned medium (hAFMSCs-CM) as an apoptotic reagent on the human MCF-7 breast cancer cell line.
Methods: Conditioned medium (CM) was prepared from hAFMSCs. After treating MCF-7 cells with CM, a number of analytical procedures (MTT, real-time PCR, western blot, and flow cytometry) were recruited to evaluate the cell viability, Bax and Bcl-2 gene expression, P53 protein expression, and apoptosis, respectively. Human fibroblast cells (Hu02) were used as the negative control. In addition, an integrated approach to meta-analysis was performed.
Results: The MCF-7 cells' viability was decreased significantly after 24 hours (P < 0.0001) and 72 hours (P < 0.05) of treatment. Compared with the control cells, Bax gene's mRNA expression increased and Bcl-2's mRNA expression decreased considerably after treating for 24 hours with 80% hAFMSCs-CM (P = 0.0012, P < 0.0001, respectively); an increasing pattern in P53 protein expression could also be observed. The flow cytometry analysis indicated apoptosis. Results from literature mining and the integrated meta-analysis showed that hAFMSCs-CM is able to activate a molecular network where Bcl2 downregulation stands in harmony with the upregulation of P53, EIF5A, DDB2, and Bax, leading to the activation of apoptosis.
Conclusion: Our finding demonstrated that hAFMSCs-CM presents apoptotic effect on MCF-7 cells; therefore, the application of hAFMSCs-CM, as a therapeutic reagent, can suppress breast cancer cells' viabilities and induce apoptosis.
BioimpactsPharmacology, Toxicology and Pharmaceutics-Pharmaceutical Science
CiteScore
4.80
自引率
7.70%
发文量
36
审稿时长
5 weeks
期刊介绍:
BioImpacts (BI) is a peer-reviewed multidisciplinary international journal, covering original research articles, reviews, commentaries, hypotheses, methodologies, and visions/reflections dealing with all aspects of biological and biomedical researches at molecular, cellular, functional and translational dimensions.