癌症干细胞样细胞在由对p53诱导的凋亡具有抗性的重组三阴性乳腺癌细胞群体进行侵袭和间充质转化过程中的作用

Role of Cancer Stem-like Cells in the Process of Invasion and Mesenchymal Transformation by a Reconstituted Triple-negative Breast Cancer Cell Population Resistant to p53-induced Apoptosis.

作者信息

Inoue Sana, Imanishi Miki, Kanzaki Ai, Fujimoto Atsumi, Maeyama Marina, Okamoto Ayaka, Matsuda Hiroka, Yoshikawa Kiyotsugu, Takahashi Rei

机构信息

Graduate School of Pharmaceutical Sciences Doshisha Women's College of Liberal Arts, Kyoto, Japan.

Faculty of Pharmaceutical Sciences Doshisha Women's College of Liberal Arts, Kyoto, Japan.

出版信息

Acta Histochem Cytochem. 2022 Oct 28;55(5):169-184. doi: 10.1267/ahc.22-00076. Epub 2022 Oct 25.

Abstract

We investigated the role of cancer stem cells (CSCs) in a population of triple-negative breast cancer (TNBC) cells that are resistant to apoptosis. A human breast cancer cell population capable of inducing p53 expression with doxycycline (Dox) was created and used as an untreated control (UT). After the addition of Dox to UT for 5 days, the cell population reconstituted with cells showing resistance to apoptosis was named RE. Fluorescence-activated cell sorting (FACS) and immunostaining revealed that after the addition of Dox, the ratio of cells in the S and G2/M phases decreased in UT as apoptosis proceeded, but did not markedly change in apoptosis-resistant RE. CSC-like cells in RE exhibited a cell morphology with a larger ratio of the major/minor axis than UT. FACS showed that RE had a higher proportion of CSC-like cells and contained more CD44CD24 mesenchymal CSCs than ALDH1A3 epithelial-like CSCs. In a Matrigel invasion assay, UT was more likely to form a three-dimensional cell population, whereas RE exhibited a planar population, higher migration ability, and the up-regulated expression of epithelial-mesenchymal transition-related genes. These results provide insights into the mechanisms by which TNBC cells acquire treatment resistance at the time of recurrence.

摘要

我们研究了癌症干细胞(CSCs)在一群对凋亡具有抗性的三阴性乳腺癌(TNBC)细胞中的作用。创建了一个能够用强力霉素(Dox)诱导p53表达的人乳腺癌细胞群体,并将其用作未处理对照(UT)。在向UT中添加Dox 5天后,用对凋亡具有抗性的细胞重建的细胞群体被命名为RE。荧光激活细胞分选(FACS)和免疫染色显示,添加Dox后,随着凋亡的进行,UT中处于S期和G2/M期的细胞比例下降,但在抗凋亡的RE中没有明显变化。RE中的CSC样细胞表现出一种细胞形态,其长轴与短轴的比例大于UT。FACS显示,RE中CSC样细胞的比例更高,并且包含更多的CD44CD24间充质CSC,而不是ALDH1A3上皮样CSC。在基质胶侵袭试验中,UT更有可能形成三维细胞群体,而RE表现出平面群体、更高的迁移能力以及上皮-间质转化相关基因的表达上调。这些结果为TNBC细胞在复发时获得治疗抗性的机制提供了见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/714f/9631983/76594ec4327d/AHC22-00076f01.jpg

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