Alanteet Alaa, Attia Hala, Alfayez Musaed, Mahmood Amer, Alsaleh Khalid, Alsanea Sary
Department of Pharmacology and Toxicology, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia.
Anatomy Department, Stem Cell Unit, College of Medicine, King Saud University, Riyadh 11451, Saudi Arabia.
Saudi Pharm J. 2024 Jan;32(1):101923. doi: 10.1016/j.jsps.2023.101923. Epub 2023 Dec 18.
This study aims to explore the anti-proliferative, pro-apoptotic, and anti-migration activities of liraglutide (LGT) in MCF-7 breast cancer (BC) cells in subjects with obesity, particularly its effects on the PI3K/Akt/mTOR/AMPK pathway. The role of AMPK/SIRT-1, an essential regulator of adipokine production, in the effect of LGT on the production of adipose-derived adipokine was also assessed. MCF-7 cells were incubated in conditioned medium (CM) generated from adipose-derived stem cells (ADSCs) of obese subjects. MCF-7 cells were then treated with LGT for 72 h. Anti-proliferative, pro-apoptotic, and anti-migration activities were investigated using alamarBlue, annexin V stain, and scratch assay, respectively. Protein levels of phosphorylated PI3K, p-Akt, p-mTOR, and p-AMPK were investigated using immunoblotting. Levels of adipokines in ADSCs were determined using RT-PCR before and after transfection of ADSCs using the specific small interference RNA sequences for AMPK and SIRT-1. LGT evoked anti-proliferative, apoptotic, and potential anti-migratory properties on MCF-7 cells incubated in CM from obese ADSCs and significantly mitigated the activity of the PI3K/Akt/mTOR survival pathway-but not AMPK-in MCF-7 cells. Furthermore, the anti-proliferative effects afforded by LGT were similar to those mediated by LY294002 (PI3K inhibitor) and rapamycin (mTOR inhibitor). Our results reveal that transfection of AMPK/SIRT-1 genes did not affect the beneficial role of LGT in the expression of adipokines in ADSCs. In conclusion, LGT elicits anti-proliferative, apoptotic, and anti-migratory effects on BC cells in obese conditions by suppressing the activity of survival pathways; however, this effect is independent of the AMPK/SIRT1 pathway in ADSCs or AMPK in BC cells.
本研究旨在探讨利拉鲁肽(LGT)在肥胖受试者的MCF-7乳腺癌(BC)细胞中的抗增殖、促凋亡和抗迁移活性,特别是其对PI3K/Akt/mTOR/AMPK信号通路的影响。同时还评估了脂肪因子产生的关键调节因子AMPK/SIRT-1在LGT对脂肪源性脂肪因子产生影响中的作用。将MCF-7细胞在肥胖受试者脂肪来源干细胞(ADSCs)产生的条件培养基(CM)中培养。然后用LGT处理MCF-7细胞72小时。分别使用alamarBlue、膜联蛋白V染色和划痕试验研究抗增殖、促凋亡和抗迁移活性。使用免疫印迹法研究磷酸化PI3K、p-Akt、p-mTOR和p-AMPK的蛋白水平。在使用针对AMPK和SIRT-1的特异性小干扰RNA序列转染ADSCs之前和之后,使用RT-PCR测定ADSCs中脂肪因子的水平。LGT对在肥胖ADSCs的CM中培养的MCF-7细胞具有抗增殖、凋亡和潜在的抗迁移特性,并显著减轻了MCF-7细胞中PI3K/Akt/mTOR存活信号通路的活性,但不影响AMPK的活性。此外,LGT提供的抗增殖作用与LY294002(PI3K抑制剂)和雷帕霉素(mTOR抑制剂)介导的作用相似。我们的结果表明,AMPK/SIRT-1基因的转染并不影响LGT对ADSCs中脂肪因子表达的有益作用。总之,LGT通过抑制存活信号通路的活性,在肥胖条件下对BC细胞产生抗增殖、凋亡和抗迁移作用;然而,这种作用独立于ADSCs中的AMPK/SIRT1信号通路或BC细胞中的AMPK。