Pancreatic cancer is the seventh leading cause of cancer-related deaths globally. Metformin is the standard first-line of treatment for hyperglycemia in Type 2 diabetes, whereas pitavastatin is a cholesterol-lowering drug used to prevent cardiovascular diseases. Both these agents evidently exert anticancer effects on pancreatic cancer; however, it remains unclear whether cotreatment using them has additive or synergistic anticancer effects on pancreatic cancer. Thus, we herein used the ASPC-1 and PANC-1 cells and treated them with metformin and/or pitavastatin. We performed the cell viability assay, transwell migration assay, and cell cycle analysis using flow cytometry. Western blotting was used to determine protein levels. We found that cotreatment with metformin (30 mM) and pitavastatin (10 μM) significantly reduced cell viability; caused G0/G1 cell cycle arrest; upregulated the expression levels of Bax, PCNA, cleaved PARP-1, cleaved caspase-3, LC3 II, and p27 ^Kip1/p21^Cip1; and inhibited cell migration. The combination index value for cell viability indicated a synergistic interaction between metformin and pitavastatin. Moreover, cotreating the cells with metformin (30 mM) and pitavastatin (10 μM) could preserve mitochondrial function, activate AMPK, and inhibit PI3K/mTOR than treatment with metformin or pitavastatin alone. These findings clearly indicated that metformin plus pitavastatin had a synergistic anticancer effect on pancreatic cancer cells, potentially caused due to the activation of AMPK and inhibition of PI3K/mTOR signaling. Altogether, our results provide that use of metformin plus pitavastatin maybe serve as a chemotherapeutic agent for human pancreatic cancer in future.

中文翻译：

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匹伐他汀和二甲双胍协同激活胰腺癌细胞的凋亡和自噬

胰腺癌是全球癌症相关死亡的第七大原因。二甲双胍是 2 型糖尿病高血糖的标准一线治疗药物，而匹伐他汀是一种用于预防心血管疾病的降胆固醇药物。这两种药物对胰腺癌都有明显的抗癌作用；然而，尚不清楚使用它们的联合治疗是否对胰腺癌具有附加或协同的抗癌作用。因此，我们在此使用了 ASPC-1 和 PANC-1 细胞并用二甲双胍和/或匹伐他汀处理它们。我们使用流式细胞术进行了细胞活力测定、transwell 迁移测定和细胞周期分析。蛋白质印迹用于确定蛋白质水平。我们发现二甲双胍 (30 mM) 和匹伐他汀 (10 μM) 共同处理显着降低了细胞活力；导致 G0/G1 细胞周期停滞；上调 Bax、PCNA、裂解的 PARP-1、裂解的 caspase-3、LC3 II 和 p27 的表达水平^Kip1 /p21 ^Cip1 ; 并抑制细胞迁移。细胞活力的组合指数值表明二甲双胍和匹伐他汀之间的协同相互作用。此外，与单独使用二甲双胍或匹伐他汀相比，用二甲双胍 (30 mM) 和匹伐他汀 (10 μM) 共同处理细胞可以保留线粒体功能、激活 AMPK 并抑制 PI3K/mTOR。这些发现清楚地表明，二甲双胍加匹伐他汀对胰腺癌细胞具有协同抗癌作用，这可能是由于 AMPK 的激活和 PI3K/mTOR 信号传导的抑制所致。总之，我们的结果表明，二甲双胍加匹伐他汀的使用可能在未来作为人类胰腺癌的化疗药物。