College of Traditional Chinese Medicine, Chongqing Medical University, #1 Yixueyuan Road, Yuzhong District, 400016, Chongqing, People's Republic of China.
Department of Hematology and Oncology, Chongqing Hospital of Traditional Chinese Medicine, Chongqing, People's Republic of China.
Med Oncol. 2021 Nov 27;39(1):16. doi: 10.1007/s12032-021-01609-4.
The present study investigates the underlying mechanisms of treatment with osthole (OST) combined with lobaplatin in human triple-negative MDA-MB-231 breast cancer cells. Human triple-negative MDA-MB-231 breast cancer cells were treated with different concentrations of OST (0.1, 1, 5, 10, 20, 50, and 100 μM) alone or in combination with 10 μM lobaplatin for 48 h. Cell viability was determined and compared between the treatment groups with the Cell Counting Kit-8 assay. Transcriptome sequencing (Project Number: M-GSGC0250521) was employed to elucidate the gene expression profile of the control group and the OST treatment group, and differentially expressed genes (DEGs) were identified based on the following criteria: logFC > 0, P < 0.05. KEGG enrichment analysis was employed to determine the biological functions of these DEGs and the related signaling pathways. Finally, flow cytometry and western blotting were used to assess differences in the apoptosis rate and protein expression in MDA-MB-231 cells subjected to different treatments. The findings showed that OST inhibited the growth of MDA-MB-231 cells in a concentration-dependent manner and cell proliferation was significantly inhibited (as indicated by a decrease of 40%) at the OST concentration of 50 μM (P < 0.05). Transcriptome sequencing identified 4712 DEGs, including 2169 upregulated DEGs and 2543 downregulated DEGs. Enrichment analysis indicated that the DEGs played a role in apoptosis, p53 signaling, DNA replication, and cell cycle. In vitro experiments showed that OST and lobaplatin could significantly induce apoptosis in the MDA-MB-231 cells (P < 0.05), as indicated by elevation in the translation level of p53/Bax/caspase-3 p17 and downregulation of the Bcl-2 protein. Finally, combined treatment with OST and lobaplatin had an enhanced anti-tumor effect (P < 0.05) on proliferation and apoptosis, as well as more obvious effects on the related proteins (p53, Bax, Bcl-2, and caspase-3 p17). Thus, OST enhanced the apoptosis-mediated growth inhibitory effect of lobaplatin on breast cancer cells and has potential for the treatment of breast cancer in the future.
本研究旨在探讨蛇床子素（OST）联合洛铂治疗人三阴性 MDA-MB-231 乳腺癌细胞的作用机制。用不同浓度的 OST（0.1、1、5、10、20、50 和 100μM）单独或联合 10μM 洛铂处理人三阴性 MDA-MB-231 乳腺癌细胞 48 小时。采用细胞计数试剂盒-8 法检测细胞活力并与各处理组进行比较。采用转录组测序（项目编号：M-GSGC0250521）阐明对照组和 OST 处理组的基因表达谱，并根据以下标准鉴定差异表达基因（DEGs）：logFC>0，P<0.05。采用 KEGG 富集分析确定这些 DEGs 的生物学功能及相关信号通路。最后，采用流式细胞术和 Western blot 法检测不同处理后 MDA-MB-231 细胞的凋亡率和蛋白表达差异。结果表明，OST 呈浓度依赖性抑制 MDA-MB-231 细胞生长，当 OST 浓度为 50μM 时细胞增殖明显抑制（下降 40%）（P<0.05）。转录组测序鉴定出 4712 个 DEGs，其中上调基因 2169 个，下调基因 2543 个。富集分析表明，DEGs 在凋亡、p53 信号转导、DNA 复制和细胞周期中发挥作用。体外实验表明，OST 和洛铂可显著诱导 MDA-MB-231 细胞凋亡（P<0.05），表现为 p53/Bax/caspase-3 p17 翻译水平升高和 Bcl-2 蛋白下调。最后，OST 和洛铂联合治疗对增殖和凋亡具有增强的抗肿瘤作用（P<0.05），并对相关蛋白（p53、Bax、Bcl-2 和 caspase-3 p17）的作用更明显。因此，OST 增强了洛铂对乳腺癌细胞凋亡介导的生长抑制作用，有望成为未来乳腺癌的治疗方法。
