QAP21抑制转移性乳腺癌细胞的干性和运动能力

doi:10.5246/jcps.2021.04.024

中国药学(英文版) ›› 2021, Vol. 30 ›› Issue (4): 289-305.DOI: 10.5246/jcps.2021.04.024

QAP21抑制转移性乳腺癌细胞的干性和运动能力

雍灵¹, 姚烨¹, 韩梦仪¹, 严晓雪², 姚庆宇¹, 郭昱辰¹, 薛钧升¹, 陈国术²^,^*(), 周田彦¹^,^*()

1. 北京大学医学部药学院分子药剂学与新释药系统北京市重点实验室, 北京 100191
2. 广州大学化学与化工学院化学基础实验室, 广东广州 510006

收稿日期:2020-05-20 修回日期:2020-07-11 接受日期:2020-08-15 出版日期:2021-04-30 发布日期:2021-04-30
通讯作者: 陈国术, 周田彦
作者简介:
+ Tel.: +86-10-82801717, E-mail: tianyanzhou@bjmu.edu.cn
+ Tel.: +86-20-39366902, E-mail: gdgschen@gzhu.edu.cn
基金资助:
Beijing Municipal Natural Science Foundation (Grant No. 7192100) and Innovation Team of Ministry of Education (Grant No. BMU2017TD003).

QAP21 reduces stemness and mobility of metastatic breast cancer cells involving D1DR activation

Ling Yong¹, Ye Yao¹, Mengyi Han¹, Xiaoxue Yan², Qingyu Yao¹, Yuchen Guo¹, Junsheng Xue¹, Guoshu Chen²^,^*(), Tianyan Zhou¹^,^*()

1 Beijing Key Laboratory of Molecular Pharmaceutics and New Drug Delivery System, Department of Pharmaceutics, School of Pharmaceutical Sciences, Peking University, Beijing 100191, China
2 Chemical Basic Laboratory, School of Chemistry and Chemical Engineering, Guangzhou University, Guangzhou 510006, China

Received:2020-05-20 Revised:2020-07-11 Accepted:2020-08-15 Online:2021-04-30 Published:2021-04-30
Contact: Guoshu Chen, Tianyan Zhou

摘要/Abstract

摘要：

乳腺癌转移是绝大多数乳腺癌患者死亡的原因, 而肿瘤干性与转移密切相关。之前研究显示可以通过激动多巴胺D1受体(dopamine D1 receptor, D1DR)来减少肿瘤干细胞(cancer stem-like cells, CSCs)。本研究旨在探索新化合物QAP21在转移性乳腺癌细胞中的药效及潜在机制。结果表明, QAP21剂量依赖性地抑制了4T1和MDA-MB-231细胞的集落形成、细胞迁移和侵袭；还明显地抑制了细胞球形成并降低了细胞中CSC比例, 说明其较好地抑制了肿瘤干性。QAP21还影响了与肿瘤转移密切相关的NF-κB/Akt/EMT通路的重要分子。此外, QAP21上调了两种细胞的D1DR表达并升高其cAMP和cGMP含量, 且D1DR特异性拮抗剂SCH 23390部分或完全拮抗了QAP21的上述作用, 均提示QAP21的效应与激动D1DR有关。总之, QAP21有效地降低了转移性乳腺癌细胞的干性和运动能力, 提示其可能用于转移性乳腺癌治疗的潜力。

关键词: 转移性乳腺癌, 多巴胺D1受体, 肿瘤干性, 细胞运动, QAP21

Abstract:

Breast cancer is the second leading cause of cancer death in women mainly due to metastasis, which is closely related to cancer stemness. Evidence has shown that cancer stem-like cells (CSCs), which are responsible for cancer stemness, can be decreased by activating dopamine D1 receptor (D1DR). In the present study, we aimed to explore the pharmacological effects as well as the underlying mechanisms of QAP21, a newly synthesized compound that can be orally administered, in metastatic breast cancer cells. Our results showed that QAP21 dose-dependently inhibited the ability of colony formation in 4T1 and MDA-MB-231 cells. Cell mobility, including cell migration and invasion, was also remarkably inhibited. Besides, QAP21 significantly inhibited mammosphere formation and decreased CSC proportion, indicating reduced cancer stemness. We further verified that the nuclear factor-kappa B (NF-κB)/Akt/epithelial-mesenchymal transition (EMT) pathway was markedly impacted by QAP21 treatment. Moreover, QAP21 up-regulated the expressions of D1DR and its second messengers, including cAMP and cGMP, which can be increased when D1DR is activated. SCH 23390, a specific D1DR antagonist, partially or completely reversed the above-mentioned effects of QAP21, indicating that D1DR activation might be involved in the underlying mechanism of QAP21. In summary, QAP21 effectively reduced breast cancer stemness and cell mobility, indicating its potential use for metastatic breast cancer therapy.

Key words: Metastatic breast cancer, Dopamine D1 receptor, Cancer stemness, Cell mobility, QAP21

Supporting:

雍灵, 姚烨, 韩梦仪, 严晓雪, 姚庆宇, 郭昱辰, 薛钧升, 陈国术, 周田彦. QAP21抑制转移性乳腺癌细胞的干性和运动能力[J]. 中国药学(英文版), 2021, 30(4): 289-305.

Ling Yong, Ye Yao, Mengyi Han, Xiaoxue Yan, Qingyu Yao, Yuchen Guo, Junsheng Xue, Guoshu Chen, Tianyan Zhou. QAP21 reduces stemness and mobility of metastatic breast cancer cells involving D1DR activation[J]. Journal of Chinese Pharmaceutical Sciences, 2021, 30(4): 289-305.

图/表 7

Figure 1. QAP21 activated D1DR in metastatic breast cancer cells 4T1. (A) The molecular structure of QAP21. The final concentrations of QAP21 and SCH were 8 and 2 μM respectively for treatment. (B and C) Green fluorescence represents D1DR expression, while blue fluorescence represents the nuclei. (D) The fluorescence intensity of D1DR in the control group was set to 100%, and the intensity in other groups in Figure 1C was calculated correspondingly. The cAMP (E) and cGMP (F) quantities of the control and QAP21 groups were measured after incubation for 15 min. Data were shown as mean ± SD (n = 3). **P < 0.01, ***P < 0.001.

Figure 2. QAP21 inhibited cell viability and colony formation of metastatic breast cancer cells. Cytotoxicity of QAP21 in 4T1 (A) and MDA-MB-231 (B) cells (n = 6). The effect of QAP21 on cell colony formation in 4T1 and MDA-MB-231 cells (C). The relative colony survival frequency of 4T1 (D) and MDA-MB-231 (E) cells after incubation using QAP21 in the absence or presence of SCH for 48 h (n = 3). Data were shown as mean ± SD (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001.

Figure 3. Effect of QAP21 on migration ability of 4T1 cells. Migration ability of 4T1 cells treated with QAP21 alone or in combination with SCH by wound-healing (A and D) and transwell migration assays (B and E). Effect of QAP21 on invasion ability of 4T1 cells investigated by transwell invasion assay (C and F). Data were shown as mean ± SD (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001.

Figure 4. Effect of QAP21 on migration ability of MDA-MB-231 cells. Migration ability of MDA-MB-231 cells treated with QAP21 alone or in combination with SCH by wound-healing (A and D) and transwell migration assays (B and E). Effect of QAP21 on invasion ability of MDA-MB-231 cells investigated by transwell invasion assay (C and F). Data were shown as mean ± SD (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001.

Figure 5. Effect of QAP21 on mammosphere formation (A) and CSC frequency (B and C) in mammospheres derived from 4T1 cells. Data were shown as mean ± SD (n = 3). *P < 0.05, **P < 0.01.

Figure 6. Effect of QAP21 on mammosphere formation (A) and CSC frequency (B and C) in mammospheres derived from MDA-MB-231 cells. Data were shown as mean ± SD (n = 3). *P < 0.05, ***P < 0.001.

Figure 7. Effect of QAP21 on expression levels of proteins related to NF-κB (A and B), Akt (C) and EMT (D) pathways in 4T1 cells. The mean value of the control group was set to 1.00, and the relative quantity was calculated for other groups.

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QAP21抑制转移性乳腺癌细胞的干性和运动能力

QAP21 reduces stemness and mobility of metastatic breast cancer cells involving D1DR activation

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