TGF-β-Induced TMEPAI Promotes Epithelial-Mesenchymal Transition in Doxorubicin-Treated Triple-Negative Breast Cancer Cells via SMAD3 and PI3K/AKT Pathway Alteration.
TGF-β-Induced TMEPAI Promotes Epithelial-Mesenchymal Transition in Doxorubicin-Treated Triple-Negative Breast Cancer Cells via SMAD3 and PI3K/AKT Pathway Alteration.
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DOI：
10.2147/bctt.s325429
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发表时间：
2021
期刊：
影响因子：
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通讯作者：
Kato M
中科院分区：
文献类型：
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作者：
Wardhani BWK;Louisa M;Watanabe Y;Setiabudy R;Kato M
上皮间质转化（EMT）和药物外排转运蛋白的过度表达已被报道导致阿霉素耐药。我们以前的研究表明，TMEPAI（跨膜前列腺雄激素诱导蛋白）衰减阿霉素在三阴性乳腺癌细胞的敏感性。然而，TMEPAI如何有助于TNBC中的多柔比星抗性仍不清楚。因此，本研究旨在阐明TMEPAI在三阴性乳腺癌细胞阿霉素耐药中的机制。我们使用BT549、三阴性细胞野生型（WT）和BT549 TMEPAI敲除。将两种细胞用TGF-β 2 ng/mL处理24小时，然后用TGF-β 2 ng/mL和多柔比星12.9 nM处理另外24小时。然后，收获细胞并计数。进一步裂解细胞并用于RT-PCR和Western印迹分析。我们测定了增殖、凋亡、EMT标记物和药物外排转运蛋白的表达水平。此外，我们研究了PI 3 K以及SMAD 3和AKT磷酸化的表达。TNBC细胞显示在TMEPAI存在下对多柔比星较不敏感。TMEPAI显示减轻凋亡标志物Bax、Bcl 2、Caspase-3和Caspase-9的mRNA表达。我们的研究结果表明，TMEPAI的存在大大放大EMT和增加阿霉素治疗后的药物外排转运蛋白的表达。此外，我们的研究结果表明，TMEPAI减少阿霉素抑制SMAD 3磷酸化的作用。TMEPAI还显示通过减少三阴性乳腺癌细胞中的PI 3 K表达和Akt磷酸化来改变多柔比星的作用。我们的研究结果表明，TMEPAI促进EMT和药物外排转运蛋白，至少部分是通过在三阴性乳腺癌细胞中将多柔比星的作用从SMAD 3磷酸化减少转移到PI 3 K/AKT抑制。
Epithelial–mesenchymal transition (EMT) and overexpression of drug efflux transporters have been reported to cause doxorubicin resistance. Our previous study indicated that TMEPAI (transmembrane prostate androgen-induced protein) attenuated doxorubicin sensitivity in triple-negative breast cancer cells. However, how TMEPAI contributes to doxorubicin resistance in TNBC remains unclear. Thus, the present study aimed to elucidate the mechanism of TMEPAI in doxorubicin resistance in triple-negative breast cancer cells. We used BT549, triple-negative cells wild type (WT), and BT549 TMEPAI knock-out. Both cells were treated with TGF-β 2 ng/mL for 24 hours, followed by TGF-β 2 ng/mL and doxorubicin 12.9 nM for another 24 hours. Afterward, the cells were harvested and counted. Cells were further lysed and used for RT-PCR and Western blot analysis. We determined the expression levels of proliferation, apoptosis, EMT markers, and drug efflux transporters. Additionally, we investigated the expressions of PI3K as well as SMAD3 and AKT phosphorylation. TNBC cells were shown to be less sensitive to doxorubicin in the presence of TMEPAI. TMEPAI was shown to alleviate the mRNA expressions of apoptosis markers: Bax, Bcl2, Caspase-3, and Caspase-9. Our results indicated that the presence of TMEPAI greatly amplifies EMT and increases drug efflux transporter expressions after doxorubicin treatment. Furthermore, our findings demonstrated that TMEPAI reduced the action of doxorubicin in inhibiting SMAD3 phosphorylation. TMEPAI was also shown to modify the effect of doxorubicin by reducing PI3K expressions and Akt phosphorylation in triple-negative breast cancer cells. Our findings indicate that TMEPAI promotes EMT and drug efflux transporters at least in part by shifting doxorubicin action from SMAD3 phosphorylation reduction to PI3K/AKT inhibition in triple-negative breast cancer cells.