Flow cytometry F analysis of DNA content cell cycle by flow cytometry

to test pharmacologic accumulation compared between cancer and normal cells, a panel of cancer cell lines and normal epithelial cell lines were treated with the aforementioned situation simultaneously. In keeping with Fig. 4A and B, AZD6244 along with API 2 effectively killed the HDAC Inhibitors cancer cells, whereas the same treatment caused little toxicity within the normal epithelial cells. Together, our findings suggest that combining AZD6244 with other medical pharmacologic agents that increase FOXO3a activity, such as API 2, can promote the efficacy of AZD6244 treatment and also sensitize AZD6244 immune cells to growth suppression. Given the that the mixture of AZD6244 and API 2 improved FOXO3a nuclear translocation, enhanced Bim supporter connection, saved Bim transcriptional activation, and sensitized AZD6244 resistant cancer cells to growth suppression and cell death, we think that FOXO3a activation is an essential aspect in treating AZD6244 resistance. The preferential killing effect in cancer cells versus normal cells might also benefit AZD6244 therapy by preventing potential negative effects in normal cells. A model depicting molecular responses toward AZD resistant and AZD sensitive cancer cells is proposed in Fig. 5B. As yet, AZD6244 has been under analysis in 21 clinical trials with about 10 different Inguinal canal cancer types including breast cancer, colon cancer, lung cancer, melanoma, kidney cancer, hepatocellular carcinoma, pancreatic cancer, ovarian cancer, acute myelogenous leukemia, and thyroid cancer in which AZD6244 has shown promising therapeutic effects specially in cancers with BRAF mutations with lower toxicity. Other MEK inhibitors such as PD 0325901 may also be proven to have promising antitumor activity in mouse models but ocular and neurologic toxicity was shown in a phase I clinical study. In Fig. 5A, the mixture of AZD6244 and API 2 in significant cell death in the five different cancer cell lines but not in the three different normal cell lines, GW9508 suggesting that AZD6244 selectively targets cancer cells and has relatively low toxicity to normal cells. AKT and ERK are commonly activated oncogenic kinases in human cancers. Curiously, both kinases target exactly the same tumor suppressor gene, FOXO3a. It had been recognized that ERK phosphorylate FOXO3a and AKT at different phosphorylation sites. Similarly, the phosphorylation of FOXO3a by these oncogenic kinases in FOXO3a translocation from the nucleus to the cytoplasm and subsequent degradation. Taxol, LY2940024, and API 2 were proven to effortlessly stop PI3K AKT pathway and activate FOXO3a nuclear translocation and activity. In our current study, we showed that inhibition of both RAS/MEK/ ERK and PI3K/AKT pathways enhances FOXO3a activity. We showed that the service of FOXO3a and its downstream gene Bim is specially important for the maximal sensitivity of cancer cells responding to AZD6244 treatment.