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人羊水间充质干细胞条件培养基对人MCF-7乳腺癌细胞系的凋亡作用

Apoptotic effects of human amniotic fluid mesenchymal stem cells conditioned medium on human MCF-7 breast cancer cell line.

作者信息

Pashaei-Asl Roghiyeh, Pashaiasl Maryam, Ebrahimie Esmaeil, Lale Ataei Maryam, Paknejad Maliheh

机构信息

Department of Clinical Biochemistry, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran.

Department of Anatomical Sciences, School of Medicine, Tabriz University of Medical Sciences.

出版信息

Bioimpacts. 2023;13(3):191-206. doi: 10.34172/bi.2022.23813. Epub 2022 Mar 30.

Abstract

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 ( < 0.0001) and 72 hours ( < 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 ( = 0.0012,  < 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.

摘要

引言

乳腺癌是女性中最常见的恶性肿瘤,具有高死亡率和化疗耐药性。研究表明间充质干细胞在癌症治疗中可能具有抑制作用。因此,本研究使用人羊水间充质干细胞条件培养基(hAFMSCs-CM)对人MCF-7乳腺癌细胞系进行凋亡诱导。

方法

从hAFMSCs制备条件培养基(CM)。用CM处理MCF-7细胞后,采用多种分析方法(MTT、实时PCR、蛋白质印迹和流式细胞术)分别评估细胞活力、Bax和Bcl-2基因表达、P53蛋白表达及细胞凋亡情况。人成纤维细胞(Hu02)用作阴性对照。此外,还进行了综合荟萃分析。

结果

处理24小时(<0.0001)和72小时(<0.05)后,MCF-7细胞活力显著降低。用80% hAFMSCs-CM处理24小时后,与对照细胞相比,Bax基因的mRNA表达增加,Bcl-2的mRNA表达显著降低(分别为=0.0012,<0.0001);P53蛋白表达也呈现增加趋势。流式细胞术分析表明细胞发生凋亡。文献挖掘和综合荟萃分析结果显示,hAFMSCs-CM能够激活一个分子网络,其中Bcl2下调与P53、EIF5A、DDB2和Bax的上调相互协调,从而导致凋亡激活。

结论

我们的研究结果表明,hAFMSCs-CM对MCF-7细胞具有凋亡作用;因此,hAFMSCs-CM作为一种治疗试剂,可抑制乳腺癌细胞活力并诱导凋亡。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d577/10329748/414ed27d5496/bi-13-191-g001.jpg

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