目的 探讨靶向CD147的小分子抑制剂在三阴性乳腺癌(TNBC)糖代谢重编程及异常增殖表型的作用及分子机制。方法 对靶向CD147抑制剂AC-73进行化学改造,获得候选改造抑制剂HA-08;应用糖代谢相关生物化学检测评估HA-08是否参与三阴性乳腺癌糖代谢重编程过程;通过WST-1细胞增殖检测验证HA-08对TNBC细胞的异常增殖的阻断功效;利用生物信息学分析结合分子生物学验证的方法阐明HA-08阻断CD147与其配体相互作用的分子机制。结果 初步改构了8个候选化合物,通过生物验证确定HA-08作为本研究对象。糖酵解相关实验表明,HA-08能够使细胞外pH水平上调,抑制TNBC乳酸释放及乳酸脱氢酶(LDH)活性,降低TNBC细胞葡萄糖摄取。增殖实验表明,HA-08可呈时间-浓度梯度依赖性抑制TNBC细胞异常增殖。免疫共沉淀实验表明,HA-08的分子机制与其阻断CD147与MCT1相互作用密切相关。结论 靶向CD147小分子抑制剂HA-08可通过阻断CD147-MCT1相互作用,进而介导TNBC糖代谢重编程,从而达到抑制TNBC细胞异常增殖的功效。
Abstract
Objective To investigate the effect and mechanism of small molecule inhibitors targeting CD147 on the abnormal proliferation of triple negative breast cancer (TNBC) by mediating reprogramming of glucose metabolism. Methods AC-73,small molecule inhibitor torgeting CD147 was chemically modified to obtain the candidate modification inhibitors HA-08 (a small molecule inhibitor targeting CD147). Biochemical tests related with glucose metabolism were used to evaluate whether HA-08 was involved in the reprogramming process of glucose metabolism in TNBC . The blocking effect of HA-08 on the abnormal proliferation of TNBC cells was verified by WST-1 cell proliferation assay. Bioinformatics analysis combined with molecular biological verification was used to elucidate the molecular mechanism of HA-08 blocking the interaction between CD147 and its ligands. Results Eight candidate compounds were modified preliminarily, and HA-08 was selected as the object of this study through biological verification. Glycolytic experiments showed that HA-08 could up-regulate the extracellular pH level, inhibit the release of TNBC lactic acid and the activity of lactate dehydrogenase (LDH), and reduce the glucose uptake of TNBC cells. Proliferation inhibition assay showed that HA-08 reduced the abnormal proliferation of TNBC cells in a time-concentration dependent manner. Co-ip assay showed that the molecular mechanism of HA-08 was closely related with its blocking of the interaction between CD147 and MCT1. Conclusions HA-08 can inhibit the abnormal proliferation of TNBC cells by blocking the interaction of CD147-MCT1, thereby mediating the reprogramming of TNBC glucose metabolism.
关键词
三阴性乳腺癌 /
CD147 /
小分子抑制剂 /
糖代谢重编程
Key words
triple-negative breast cancer /
CD147 /
small molecule inhibitor /
reprogramming of glucose metabolism
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基金
国家自然科学基金青年项目(81703003);空军特色医学中心科技助推计划(2020KTC03);空军特色医学中心青年博士助推课题(21ZT01;21ZT07)
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