Division of Endocrinology and Metabolism, Graduate School of Medicine, Gunma University, 3-39-15 Showa-machi, Maebashi, Gunma 371-8511, Japan.
Department of Human Pathology, Graduate School of Medicine, Gunma University, 3-39-15 Showa-machi, Maebashi, Gunma 371-8511, Japan.
Molecules. 2020 Jan 23;25(3):495. doi: 10.3390/molecules25030495.
Dapagliflozin, empagliflozin, tofogliflozin, selective inhibitors of sodium-glucose cotransporter 2 (SGLT2), is used clinically to reduce circulation glucose levels in patients with type 2 diabetes mellitus by blocking the reabsorption of glucose by the kidneys. Dapagliflozin is metabolized and inactivated by UGT1A9. Empagliflozin is metabolized and inactivated by UGT1A9 and by other related isoforms UGT2B7, UGT1A3, and UGT1A8. Tofogliflozin is metabolized and inactivated by five different enzymes CYP2C18, CYP3A4, CYP3A5, CYP4A11, and CYP4F3. Dapagliflozin treatment of HCT116 cells, which express SGLT2 but not UGT1A9, results in the loss of cell adhesion, whereas HepG2 cells, which express both SGLT2 and UGT1A9, are resistant to the adhesion-related effects of dapagliflozin. PANC-1 and H1792 cells, which do not express either SGLT2 or UGT1A9, are also resistant to adhesion related effects of dapagliflozin. On the other hand, either empagliflozin or tofogliflozin treatment of HCT116, HepG2, PANC-1, and H1792 cells are resistant to the adhesion-related effects as observed in dapagliflozin treated HCT116 cells. Knockdown of UGT1A9 by shRNA in HepG2 cells increased dapagliflozin sensitivity, whereas the overexpression of UGT1A9 in HCT116 cells protected against dapagliflozin-dependent loos of cell adhesion. Dapagliflozin treatment had no effect on cellular interactions with fibronectin, vitronectin, or laminin, but it induced a loss of interaction with collagen I and IV. In parallel, dapagliflozin treatment reduced protein levels of the full-length discoidin domain receptor I (DDR1), concomitant with appearance of DDR1 cleavage products and ectodomain shedding of DDR1. In line with these observations, unmetabolized dapagliflozin increased ADAM10 activity. Dapagliflozin treatment also significantly reduced Y792 tyrosine phosphorylation of DDR1 leading to decrement of DDR1 function and detachment of cancer cells. Concomitant with these lines of results, we experienced that CEA in patients with colon cancer, which express SGLT2 but not UGT1A9, and type 2 diabetes mellitus treated by dapagliflozin in addition to chemotherapy was decreased (case 1). CEA in patients with colon cancer, which express SGLT2 but not UGT1A9, and type 2 diabetes mellitus was treated by dapagliflozin alone after radiation therapy was decreased but started to rise after cessation of dapagliflozin (case 2). CA19-9 in two of patients with pancreatic cancer and type 2 diabetes mellitus was resistant to the combination therapy of dapagliflozin and chemotherapy (case 3 and 4 respectively). PIVKAII in patients with liver cancer and type 2 diabetes mellitus, and CYFRA in patients with squamous lung cancer and type 2 diabetes mellitus was also resistant the combination therapy of dapagliflozin and chemotherapy (case 5 and 6 respectively). Taken together, these data suggest a potential role for dapagliflozin anticancer therapy against colon cancer cells that express SGLT2, but not UGT1A9.
达格列净、恩格列净、托格列净是钠-葡萄糖协同转运蛋白 2(SGLT2)的选择性抑制剂,临床上用于通过阻断肾脏对葡萄糖的重吸收来降低 2 型糖尿病患者的循环葡萄糖水平。达格列净由 UGT1A9 代谢和失活。恩格列净由 UGT1A9 和其他相关同工酶 UGT2B7、UGT1A3 和 UGT1A8 代谢和失活。托格列净由五种不同的酶 CYP2C18、CYP3A4、CYP3A5、CYP4A11 和 CYP4F3 代谢和失活。达格列净处理表达 SGLT2 但不表达 UGT1A9 的 HCT116 细胞会导致细胞黏附丧失,而表达 SGLT2 和 UGT1A9 的 HepG2 细胞对达格列净的黏附相关作用具有抗性。既不表达 SGLT2 也不表达 UGT1A9 的 PANC-1 和 H1792 细胞也对达格列净的黏附相关作用具有抗性。另一方面,恩格列净或托格列净处理 HCT116、HepG2、PANC-1 和 H1792 细胞均能抵抗达格列净处理 HCT116 细胞所观察到的黏附相关作用。在 HepG2 细胞中用 shRNA 敲低 UGT1A9 增加了达格列净的敏感性,而在 HCT116 细胞中过表达 UGT1A9 则可防止达格列净依赖性细胞黏附丧失。达格列净处理对细胞与纤维连接蛋白、纤连蛋白或层粘连蛋白的相互作用没有影响,但它诱导与胶原蛋白 I 和 IV 的相互作用丧失。与此平行,达格列净处理降低了全长盘状结构域受体 I(DDR1)的蛋白水平,同时出现 DDR1 切割产物和 DDR1 外显子脱落。与这些观察结果一致,未代谢的达格列净增加了 ADAM10 的活性。达格列净处理还显著降低了 Y792 酪氨酸磷酸化的 DDR1,导致 DDR1 功能下降和癌细胞脱落。与这些结果一致,我们发现结肠癌患者的 CEA(表达 SGLT2 但不表达 UGT1A9)和接受达格列净加化疗治疗的 2 型糖尿病患者的 CEA(病例 1)降低。结肠癌患者的 CEA(表达 SGLT2 但不表达 UGT1A9)和接受达格列净加放疗治疗的 2 型糖尿病患者的 CEA 降低,但在停止达格列净治疗后开始升高(病例 2)。两名胰腺癌和 2 型糖尿病患者的 CA19-9 对达格列净和化疗的联合治疗有抗性(病例 3 和 4 分别)。两名肝癌和 2 型糖尿病患者的 PIVKAII 和两名鳞状肺癌和 2 型糖尿病患者的 CYFRA 对达格列净和化疗的联合治疗也有抗性(病例 5 和 6 分别)。总的来说,这些数据表明达格列净可能对表达 SGLT2 但不表达 UGT1A9 的结肠癌细胞具有抗癌治疗作用。