{"title":"基于三维培养乳腺癌细胞单细胞RNA测序的共培养细胞与纯培养细胞对顺铂的差异反应","authors":"Shuqing Yang, Peixian Chen, Xiaofan Mao, KaiRong Lin, Wei Li, Tiancheng He, Huiqi Huang, AiGuo Wu, Wei Luo, Guolin Ye, Guangyu Yao, Dan Zhou","doi":"10.31083/j.fbl2912406","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>The current study aimed to develop an experimental approach for the direct co-culture of three-dimensional breast cancer cells using single-cell RNA sequencing (scRNA-seq).</p><p><strong>Methods: </strong>The following four cell culture groups were established in the Matrigel matrix: the untreated Michigan Cancer Foundation (MCF)-7 cell culture group, the MCF-7 cell culture plus cisplatin group, the untreated co-culture group, and the cell co-culture plus cisplatin group. For cell co-culture, MCF-7 cells, human mammary fibroblasts, and human umbilical vein endothelial cells were mixed at a ratio of 1:1:1. Cisplatin was applied at a concentration of 1.25 μg/mL, and the cells were harvested after 2 days and subjected to scRNA-seq. Data were analyzed using a single-cell RNA sequencing data analysis pipeline with R language.</p><p><strong>Results: </strong>The response of MCF-7 cells to cisplatin differed among the four groups. The transcriptomic response of MCF-7 cells to cisplatin in the co-culture model was not as significant as that in the mono-culture model. Moreover, the pathways related to apoptosis, DNA damage, hypoxia, and metastasis in the co-culture groups were enriched in the genes that were differentially expressed based on cisplatin treatment.</p><p><strong>Conclusion: </strong>scRNA-seq analysis revealed that the response of MCF-7 cells to cisplatin in the co-culture model was lower than that in the mono-culture model. Therefore, the three-dimensional cell co-culture model can be applied to tumor research to better mimic the pathophysiological environment <i>in vivo</i> and can be a well-modified research method.</p>","PeriodicalId":73069,"journal":{"name":"Frontiers in bioscience (Landmark edition)","volume":"29 12","pages":"406"},"PeriodicalIF":3.3000,"publicationDate":"2024-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Differential Response to Cisplatin between Co-cultured Cells and Pure Cultured Cells Based on Single-cell RNA Sequencing of Three-dimensional-cultured Breast Cancer Cells.\",\"authors\":\"Shuqing Yang, Peixian Chen, Xiaofan Mao, KaiRong Lin, Wei Li, Tiancheng He, Huiqi Huang, AiGuo Wu, Wei Luo, Guolin Ye, Guangyu Yao, Dan Zhou\",\"doi\":\"10.31083/j.fbl2912406\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Objective: </strong>The current study aimed to develop an experimental approach for the direct co-culture of three-dimensional breast cancer cells using single-cell RNA sequencing (scRNA-seq).</p><p><strong>Methods: </strong>The following four cell culture groups were established in the Matrigel matrix: the untreated Michigan Cancer Foundation (MCF)-7 cell culture group, the MCF-7 cell culture plus cisplatin group, the untreated co-culture group, and the cell co-culture plus cisplatin group. For cell co-culture, MCF-7 cells, human mammary fibroblasts, and human umbilical vein endothelial cells were mixed at a ratio of 1:1:1. Cisplatin was applied at a concentration of 1.25 μg/mL, and the cells were harvested after 2 days and subjected to scRNA-seq. Data were analyzed using a single-cell RNA sequencing data analysis pipeline with R language.</p><p><strong>Results: </strong>The response of MCF-7 cells to cisplatin differed among the four groups. The transcriptomic response of MCF-7 cells to cisplatin in the co-culture model was not as significant as that in the mono-culture model. Moreover, the pathways related to apoptosis, DNA damage, hypoxia, and metastasis in the co-culture groups were enriched in the genes that were differentially expressed based on cisplatin treatment.</p><p><strong>Conclusion: </strong>scRNA-seq analysis revealed that the response of MCF-7 cells to cisplatin in the co-culture model was lower than that in the mono-culture model. Therefore, the three-dimensional cell co-culture model can be applied to tumor research to better mimic the pathophysiological environment <i>in vivo</i> and can be a well-modified research method.</p>\",\"PeriodicalId\":73069,\"journal\":{\"name\":\"Frontiers in bioscience (Landmark edition)\",\"volume\":\"29 12\",\"pages\":\"406\"},\"PeriodicalIF\":3.3000,\"publicationDate\":\"2024-11-29\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Frontiers in bioscience (Landmark edition)\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.31083/j.fbl2912406\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Frontiers in bioscience (Landmark edition)","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.31083/j.fbl2912406","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
目的:本研究旨在建立一种利用单细胞RNA测序(scRNA-seq)直接共培养三维乳腺癌细胞的实验方法。方法:在Matrigel基质中建立4个细胞培养组:未经处理的Michigan Cancer Foundation (MCF)-7细胞培养组、MCF-7细胞培养+顺铂组、未经处理的共培养组、细胞共培养+顺铂组。细胞共培养时,将MCF-7细胞、人乳腺成纤维细胞和人脐静脉内皮细胞按1:1:1的比例混合。顺铂浓度为1.25 μg/mL, 2天后收获细胞,进行scrna测序。数据分析使用单细胞RNA测序数据分析管道与R语言。结果:四组间MCF-7细胞对顺铂的反应存在差异。在共培养模型中,MCF-7细胞对顺铂的转录组反应不如单培养模型显著。此外,在共培养组中,与细胞凋亡、DNA损伤、缺氧和转移相关的通路在顺铂治疗差异表达的基因中富集。结论:scRNA-seq分析显示,共培养模型中MCF-7细胞对顺铂的反应低于单培养模型。因此,三维细胞共培养模型可以应用于肿瘤研究,更好地模拟体内病理生理环境,是一种完善的研究方法。
Differential Response to Cisplatin between Co-cultured Cells and Pure Cultured Cells Based on Single-cell RNA Sequencing of Three-dimensional-cultured Breast Cancer Cells.
Objective: The current study aimed to develop an experimental approach for the direct co-culture of three-dimensional breast cancer cells using single-cell RNA sequencing (scRNA-seq).
Methods: The following four cell culture groups were established in the Matrigel matrix: the untreated Michigan Cancer Foundation (MCF)-7 cell culture group, the MCF-7 cell culture plus cisplatin group, the untreated co-culture group, and the cell co-culture plus cisplatin group. For cell co-culture, MCF-7 cells, human mammary fibroblasts, and human umbilical vein endothelial cells were mixed at a ratio of 1:1:1. Cisplatin was applied at a concentration of 1.25 μg/mL, and the cells were harvested after 2 days and subjected to scRNA-seq. Data were analyzed using a single-cell RNA sequencing data analysis pipeline with R language.
Results: The response of MCF-7 cells to cisplatin differed among the four groups. The transcriptomic response of MCF-7 cells to cisplatin in the co-culture model was not as significant as that in the mono-culture model. Moreover, the pathways related to apoptosis, DNA damage, hypoxia, and metastasis in the co-culture groups were enriched in the genes that were differentially expressed based on cisplatin treatment.
Conclusion: scRNA-seq analysis revealed that the response of MCF-7 cells to cisplatin in the co-culture model was lower than that in the mono-culture model. Therefore, the three-dimensional cell co-culture model can be applied to tumor research to better mimic the pathophysiological environment in vivo and can be a well-modified research method.