Department of Throat and Breast Surgery, The Second Affiliated Hospital of Soochow University, Soochow P.R. China.
Department of Throat and Breast Surgery, Affiliated Hospital of Yangzhou University, Yangzhou P.R. China.
Cancer Biol Ther. 2021 Sep 2;22(7-9):465-477. doi: 10.1080/15384047.2021.1949228. Epub 2021 Sep 5.
Cisplatin is an effective chemotherapeutic agent in facilitating the inhibition of proliferation, migration, and invasion in cancerous cells. However, the detailed mechanism of the regulation by cisplatin of human breast cancer cells is still unclear. This study aimed to investigate the mechanism of kruppel-like factor 2 (KLF2) transcription factor in cisplatin therapy for breast cancer. RT-qPCR was performed to quantify the expression of KLF2 and WEE1 in clinical tissue samples from breast cancer patients and in MDA-MB-231 cells. ChIP assay and dual-luciferase reporter assay were used to analyze the potential-binding sites of KLF2 and WEE1 promoter. Gain- or loss-of-function approaches were used to manipulate KLF2 and WEE1 in cisplatin-treated MDA-MB-231 cells, and the mechanism of KLF2 in breast cancer was evaluated both CCK-8 assay, flow cytometry, Transwell assay, and Western blot. Further validation of the KLF2 was performed on nude mouse models. Breast cancer tissues and cells showed a relative decline of KLF2 expression and abundant WEE1 expression. Cisplatin inhibited the proliferation, migration, and invasion of MDA-MB-231 cells. Overexpression of KLF2 enhanced the inhibitory effect of cisplatin on the malignant characteristics of MDA-MB-231 cells . KLF2 targeted WEE1 and negatively regulated its expression, thus enhancing the sensitivity to cisplatin of breast cancer cells as well as tumor-bearing mice. Overall, these results suggest that KLF2 can potentially inhibit WEE1 expression and sensitize breast cancer cells to cisplatin, thus presenting a promising adjunct treatment.
顺铂是一种有效的化疗药物，可促进癌细胞增殖、迁移和侵袭的抑制。然而，顺铂调节人乳腺癌细胞的详细机制仍不清楚。本研究旨在探讨 Kruppel 样因子 2（KLF2）转录因子在顺铂治疗乳腺癌中的作用机制。采用 RT-qPCR 定量检测乳腺癌患者临床组织样本和 MDA-MB-231 细胞中 KLF2 和 WEE1 的表达。采用 ChIP assay 和双荧光素酶报告基因检测分析 KLF2 和 WEE1 启动子的潜在结合位点。采用 gain-或 loss-of-function 方法在顺铂处理的 MDA-MB-231 细胞中操纵 KLF2 和 WEE1，并用 CCK-8 assay、流式细胞术、Transwell assay 和 Western blot 评估 KLF2 在乳腺癌中的作用机制。在裸鼠模型中进一步验证 KLF2。乳腺癌组织和细胞显示 KLF2 表达相对下降和 WEE1 表达丰富。顺铂抑制 MDA-MB-231 细胞的增殖、迁移和侵袭。KLF2 的过表达增强了顺铂对 MDA-MB-231 细胞恶性特征的抑制作用。KLF2 靶向 WEE1 并负调控其表达，从而增强乳腺癌细胞对顺铂的敏感性以及荷瘤小鼠的敏感性。总之，这些结果表明 KLF2 可能抑制 WEE1 表达并使乳腺癌细胞对顺铂敏感，从而为联合治疗提供了一种有前途的方法。
