An aldehyde dehydrogenase 1A3 inhibitor attenuates the metastasis of human colorectal cancer
An aldehyde dehydrogenase 1A3 inhibitor attenuates the metastasis of human colorectal cancer
复制标题
乙醛脱氢酶 1A3 抑制剂可减弱人结直肠癌的转移
DOI：
10.1016/j.canlet.2022.215662
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发表时间：
2022-04-07
期刊：
影响因子：
9.7
通讯作者：
Yu, Shi-Cang
中科院分区：
文献类型：
--
作者：
Duan, Jiang-Jie;Cai, Jiao;Yu, Shi-Cang
转移是结直肠癌（CRC）患者死亡的主要原因。开发能够选择性抑制CRC细胞播散所涉及的生物学过程的治疗方案非常重要。我们使用多个Affymetrix DNA微阵列杂交数据集来识别与转移相关且对CRC患者具有显著预后价值的基因。采用实时定量PCR、免疫荧光和免疫组化染色来评估mRNA和蛋白质的表达。通过进行Transwell实验和动物实验来评估醛脱氢酶1A3（ALDH1A3）在侵袭中的作用。采用实时定量PCR、荧光素酶报告基因实验和蛋白质印迹法来识别受ALDH1A3调控的基因。采用分子对接、MTS实验、细胞热位移实验、等温滴定量热法、微量热泳动和酶活性实验来筛选和验证ALDH1A3特异性抑制剂YD1701（二苯并 - 30 - 冠 - 10 - 醚）的功效。最后，建立皮下或原位异种移植模型来研究YD1701的治疗潜力。人类ALDH1A3被确定与CRC细胞的转移表型以及患者不良预后相关。此外，ALDH1A3通过抑制miR - 200家族成员上调ZEB1和SNAI2的表达。筛选出ALDH1A3特异性抑制剂YD1701，它在体外减弱了CRC细胞的侵袭，并延长了皮下或原位异种移植小鼠的生存期。我们的结果表明，ALDH1A3通过miR - 200 - ZEB1/SANI2轴促进侵袭和转移，因此是预测CRC进展的一个合理标志物。用所鉴定的化合物YD1701抑制ALDH1A3可能是一种防止CRC转移的有效治疗方法。
Metastasis is the leading cause of death for patients with colorectal cancer (CRC). The development of therapeutic regimens that selectively inhibit the biological processes involved in CRC cell dissemination is important. We used multiple Affymetrix DNA microarray hybridization datasets to identify genes related to metastasis and have significant prognostic value for patients with CRC. Quantitative real-time PCR, immunofluorescent and immunohistochemical staining were used to evaluate mRNA and protein expression. The function of aldehyde dehydrogenase 1A3 (ALDH1A3) in invasion was assessed by performing transwell assays and animal experiments. Real-time PCR, luciferase reporter assays, and western blotting were used to identify the genes regulated by ALDH1A3. Molecular docking, MTS assays, cellular thermal shift assays, isothermal titration calorimetry, microscale thermophoresis, and enzymatic activity assays were used to screen and verify the efficacy of the ALDH1A3-specific inhibitor YD1701 (dibenzo-30-crown10-ether). Finally, subcutaneous or orthotopic xenograft models were established to investigate the therapeutic potential of YD1701. Human ALDH1A3 was identified to correlate with a metastatic phenotype in CRC cells and a poor patient prognosis. Moreover, ALDH1A3 upregulated the expression of ZEB1 and SNAI2 by inhibiting miR-200 family members. The ALDH1A3-specific inhibitor YD1701 was screened, attenuated the invasion of CRC cells in vitro, and prolonged the survival of mice bearing subcutaneous or orthotopic xenografts. Our results show that ALDH1A3 promotes invasion and metastasis via the miR-200-ZEB1/SANI2 axis and is thus a plausible marker for predicting CRC progression. Inhibiting ALDH1A3 with the identified compound YD1701 might represent an effective therapeutic approach to prevent the metastasis of CRC.