Investigation of the relationship between ruxolitinib and MK 2206's estrogen signaling pathway on PI3K/AKT ve JAK/STAT signaling pathways in BT474 and MDA-MB-231 breast cancer cell line

In a tumor cell,the imbalance of pro-apoptotic and anti-apoptotic proteins occurs. PI3K/AKT,JAK/STAT,and MAPK signaling play an essential role in the proliferation of cancer cells and the regulation of angiogenesis and metastasis. Reducing cell proliferation and stimulating cell death with inhibitors targeting these signaling pathways are therapeutic approaches. Ruxolitinib is RTK inhibitor that targets JAK1/2. MK-2206 is selective AKT inhibitor. There is a need for Ruxolitinib to act more effectively on cancer cells,to selectively increase drug accumulation in tumor cells,and to reduce the toxic and side effects of drugs. In this study,the combined treatment of Ruxolitinib and MK-2206,which have synergistic effects on BT474(ER+,PR+,HER2+) and MDA-MB-231(ER-,PR-,HER2-) breast cancer cells,aimed firstly to eliminate this deficiency. It was aimed secondly to eliminate this deficiency by synthesizing PCL nanoparticles loaded with Ruxolitinib and MK-2206. IC50 values of free and nano forms of drugs were determined by MTT;anti-proliferative effects by Survival,Soft Agar,Colony Formation,Wound healing tests; Morphological changes in cells were examined with Hoechst33342. The effects of drugs on apoptosis,ERα,MAPK,PI3K/AKT and JAK/STAT pathways in cells were investigated by immunoblotting. It was observed that Ruxolitinib+MK-2206 application activates the extrinsic pathway by inhibiting PI3K/AKT,MAPK and JAK/STAT3 pathways in BT474 cells, and activates the intrinsic pathway by inhibiting PI3K/AKT,MAPK and JAK/STAT5 pathways in MDA-MB-231 cells. It was observed that PCL-NPs didn't have toxic effect on both cells. It was observed that the IC50 of Ruxolitinib decreased 50-fold in BT474 cells with its nanoform. It has been observed that Rux-PCL-NP and MK-PCL-NP have similar effects on the PI3K/AKT,MAPK,JAK/STAT pathways in cells.