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Objective: To investigate whether licorice extracts (LE) combined with iron death inducer Erastin can induce iron death of colorectal cancer cell DLD1 through dipeptidyl peptidase-4 (DPP4).
Methods: DLD1 cells were randomly divided into control group, L-LE group (50 μg·mL-1 LE), H-LE group (100 μg·mL-1 LE), H-LE+Erastin group (100 μg·mL-1 LE+30 nmol·L-1 Erastin) and H-LE+Erastin+Sita (DPP4 inhibitor) group (100 μg·mL-1 LE+30 nmol·L-1 Erastin+31.25 μg·mL-1 Sita), with drug interventions for 24 h. Cell cloning assay and Edu-594 cell proliferation test were conducted to determine the cloning and proliferation abilities of the cells. ELISA assay was used to detect the Fe2+ content and DPP4 activity. Flow cytometry was used to detect reactive oxygen species(ROS) content. Western blotting was used to detect the expression of solute carrier family 7 member 11(SLC7A11) and arachidonate lipoxygenase 3(ALOXE3) proteins.
Results: Compared with control group, the cell cloning number decreased while the ROS content increased in L-LE group, H-LE group and H-LE+Erastin group (P<0.05 or P<0.01); and the proliferation ability and Fe2+ content decreased, DPP4 activity and ALOXE3 protein expression increased, SLC7A11 protein expression decreased in H-LE and H-LE+Erastin groups (P<0.05 or P<0.01). Compared with H-LE+Erastin group, the addition of Sita significantly increased cell cloning, proliferation and SLC7A11 protein expression, while significantly decreased Fe2+ content, DPP4 activity, ROS content and ALOXE3 protein expression (P<0.01).
Conclusion: LE combined with Erastin can induce iron death of colorectal cancer (CRC) cell through DPP4.
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目的： 探究甘草水提物（licorice extracts, LE）联合铁死亡诱导剂Erastin能否通过二肽基肽酶4（dipeptidyl peptidase-4, DPP4）诱导人结直肠腺癌上皮细胞（DLD-1）发生铁死亡。
方法： 将DLD-1细胞随机分为Control组、L-LE组（50 μg·mL-1 LE）、H-LE组（100 μg·mL-1 LE）、H-LE+Erastin组（100 μg·mL-1 LE+30 nmol·L-1 Erastin）、H-LE+Erastin+Sita（DDP4抑制剂）组（100 μg·mL-1 LE+30 nmol·L-1 Erastin+31.25 μg·mL-1 Sita），药物干预24 h。行细胞克隆实验和EDU-594细胞增殖实验测定细胞克隆及增殖情况，ELISA检测细胞Fe2+含量及DPP4活性变化，流式细胞术检测细胞活性氧（reactive oxygen species, ROS）含量，Western blotting检测细胞中溶质载体家族7成员11（solute carrier family 7 member 11, SLC7A11）、花生四烯酸脂加氢酶3（arachidonate lipoxygenase 3, ALOXE3）蛋白的表达情况。
结果： 相较于Control组，L-LE，H-LE及H-LE+Erastin处理组细胞的克隆数下降，ROS含量增加（P<0.05或P<0.01）；H-LE和H-LE+Erastin处理组细胞的增殖能力降低，Fe2+含量、DPP4活性以及ALOXE3蛋白表达升高，SLC7A11蛋白表达水平降低（P<0.05或P<0.01）。与H-LE+Erastin组相比，加入Sita处理后，细胞的克隆数量、增殖能力及SLC7A11蛋白表达显著增加，而Fe2+含量、DPP4活性、ROS生成量及ALOXE3蛋白表达水平显著降低（P<0.01）。
结论： LE联合Erastin可通过DPP4诱导结直肠癌（CRC）细胞发生铁死亡。
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陈吉，女，主管药师，主要从事临床药学、药物药效药理等药学研究。E-mail: chenjihzsds@163.com。
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陈吉，女，主管药师，主要从事临床药学、药物药效药理等药学研究。E-mail: chenjihzsds@163.com。
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