ABSTRACT
Objective: To investigate the effects of lysine acetyltransferase (Kat5) on the proliferation and apoptosis of thyroid papillary carcinoma cells. Methods: RT-PCR and Western blotting methods were used to determine the expression levels of Kat5 mRNA and protein in the thyroid papillary carcinoma BHP10-3. TPC-1 . K1 cells and thyroid epithelial Nthy-ori3-l cells. The thyroid papillary carcinoma TPC-1 cells were divided into blank control group, negative control group and silencing Kat5 group (si-Kat5 group). The cells in blank control group were not transfected. the cells in negative control group were transfected with negative control siRNA. and the cells in si-Kat5 group were transfected with Kat5 siRNA. The expression levels of Kat5 mRNA and protein in the thyroid papillary carcinoma TPC-1 cells in various groups were detected by RT-PCR and Western blotting methods. The proliferation activities of the TPC-1 cells in various groups were measured by CCK-8 method, the colony formation assay was used to measure the clone formation rates of the TPC-1 cells in various groups, the apoptotic rates of the TPC-1 cells in various groups were detected by flow cytometry, and the expression levels of cyclin-dependent kinase 2 (CDK2). P21, P53. cleaved caspase-3. P13K, phosphorylated P13K (p-P13K), AKT, and phosphorylated AKT (p-AKT) protein in the TPC-1 cells in various groups were detected by Western blotting method. Results: The expression levels of Kat5 mRNA and protein in the thyroid papillary carcinoma BHP10-3. TPC-1 and K1 cells were higher than those in thyroid epithelial Nthy-ori3-l cells (P<0. 05). The differences in the Kat5 mRNA and protein expression levels, the proliferation activities, the clone formation rates, the apoptotic rates and the expression levels of CDK2. P21. P53. cleaved caspase-3. P13K. p-P13K . AKT and p-AKT proteins in the TPCM cells between various groups were statistically significant (P<0.05). Compared with blank control group and negative control group, the expression levels of Kat5 mRNA and protein in the TPC-1 cells in si-Kat5 group were decreased ( P
ABSTRACT
Objective To explore the effects of phosphatidylinositol-3-kinase (PI3K)/serine-threonine kinase (AKT) signal pathway inhibitor LY294002 on retinal neovascularization (RNV) in oxygen-induced retinopathy (OIR) mice.Methods Totally 60 C57BL/6J mice were collected and randomly divided into the experimental group and control group,with 30 mice in each group.Then OIR model was induced by Smith methods.Rats in the experimental group were intravitreally injected with 0.5 μL LY294002,while the control group was given the same amount of phosphate buffer saline (PBS) one day before out of the incubator.Retinal sections with HE staining were applied to count the number of neovascular cell nuclei breaking through the inner limiting membrane,as well as the protein and mRNA expression of pAKT and vascular endothelial growth factor (VEGF) were detected by immunohistochemistry and RT-PCR.Results The number of retinal neovascular cell nucleus in the experimental group was obviously smaller than that in the control group (P ＜ 0.05).The protein expression of pAKT and VEGF was weakly expressed,and the absorbance (A) value of the positive cells was decreased in the experimental group compared with the control group (all P ＜ 0.05).The mRNA expression of AKT and VEGF was obviously decreased in the experimental group compared with the control group (all P ＜ 0.05).Conclusion The development of RNV in OIR mice can be markedly inhibited by LY294002 inhibiting PI3K/AKT pathway,and therefore LY294002 is expected to be an effective method for preventing vascular proliferative retinopathy.