Page 74 - 《中国药科大学学报》2026年第1期

学报
68 Journal of China Pharmaceutical University 2026, 57(1): 68 − 77

五氟利多靶向 HSPA6 抑制黑色素瘤生长的作用研究

葛 姮，姜 盼，张 馨，潘钊海 ，李德芳 **
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（滨州医学院中医学院, 山东省高等学校中药活性成分生物合成与靶点发现特色实验室, 烟台 264003）

摘 要 基于药物亲和力反应靶点稳定性（drug affinity responsive target stability, DARTS）技术解析抗精神病药物五氟利多
抗黑色素瘤的分子机制，并揭示其通过 HSPA6/p53/p21 信号轴发挥作用的生物学通路。MTT 实验和细胞集落形成能力等实
验证实五氟利多可显著下调黑色素瘤 A375 和 B16 细胞内 cyclin D1 和 CDK4 表达，诱导细胞发生 G 1 期阻滞，进而抑制黑色
素瘤细胞增殖。同时，蛋白免疫印迹、Hoechst 33342 染色和 Annexin V-FITC/PI 双染等实验结果发现，五氟利多可显著下调
细胞内 Bcl-2 的表达，上调 Bax 和 cleaved caspase-3 的表达，诱导细胞发生凋亡。进一步利用 DARTS 技术鉴定热休克蛋白
A6（HSPA6）为五氟利多结合的关键靶点，五氟利多通过上调 HSPA6 并激活 p53/p21 通路；敲低 HSPA6 不仅可逆转五氟利
多介导的 p53/p21 通路的激活，还可逆转五氟利多介导的周期阻滞和凋亡。荷瘤小鼠动物实验也证实了敲低 HSPA6 可逆转
五氟利多的体内抗肿瘤活性。因此，本研究阐明了五氟利多可通过靶向 HSPA6 激活 p53/p21 信号轴来抑制黑色素瘤进展，
这为抗精神病药物治疗肿瘤再定位提供了新视角。
关键词 黑色素瘤；五氟利多；周期阻滞；凋亡；HSPA6
中图分类号 R965;R739.5 文献标志码 A 文章编号 1000−5048(2026)01−0068−10
doi：10.11665/j.issn.1000−5048.2025041801

引用本文 葛姮，姜盼，张馨，等. 五氟利多靶向 HSPA6 抑制黑色素瘤生长的作用研究 [J]. 中国药科大学学报，2026，57（1）：68-77.

Cite this article as: GE Heng, JIANG Pan, ZHANG Xin, et al. Inhibitory effect of penfluridol on melanoma growth by targeting HSPA6[J]. J
China Pharm Univ, 2026, 57(1): 68-77.

Inhibitory effect of penfluridol on melanoma growth by targeting HSPA6
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GE Heng, JIANG Pan, ZHANG Xin, PAN Zhaohai , LI Defang **
Specialized Laboratory for Biosynthesis and Target Discovery of Active Components of Traditional Chinese Medicine in Higher
Education Institutions in Shandong Province, School of Traditional Chinese Medicine, Binzhou Medical University, Yantai 264003,
China

Abstract This study employed the drug affinity responsive target stability (DARTS) technique to investigate
the molecular mechanism of the antipsychotic drug penfluridol against melanoma, revealing the biological
pathway to exert its effect on the HSPA6/p53/p21 signaling axis. Experiments such as the
methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay and cell colony formation ability assay confirmed that
penfluridol could significantly downregulate the expression of cyclin D1 and cyclin-dependent kinase 4 (CDK4)
in melanoma A375 and B16 cells, induce cell cycle arrest in the G phase, and thus inhibit the proliferation of
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melanoma cells. Meanwhile, the results of Western blot, Hoechst 33342 staining and Annexin V-FITC/PI double
staining experiments showed that penfluridol could significantly downregulate the expression of Bcl-2 and
upregulate the expression of Bax and cleaved caspase-3, inducing cell apoptosis. Further, the DARTS technique
was used to identify heat shock 70 kD protein 6 (HSPA6) as the key target bound by penfluridol. Penfluridol
activates the p53/p21 pathway by upregulating HSPA6. Knocking down HSPA6 reverses not only the activation
of the p53/p21 pathway mediated by penfluridol but also the associated cell cycle arrest and apoptosis. Animal
experiments on tumor-bearing mice also confirmed that knocking down HSPA6 could reverse the in vivo anti-
tumor activity of penfluridol. This study clarified that penfluridol can inhibit the progression of melanoma by

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收稿日期 2025-04-18 通信作者 E-mail：pzh_bzmu@163.com
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E-mail：lidefang@163.com
基金项目 国家中医药管理局科技司-山东省卫生健康委员会共建中医药科技项目（GZY-KJS-SD-2023-094）