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AIM: To investigate the mechanism of acquired resistance remodeling to DDP (named cisplatin) by comparing the level of autophagy between the parental and DDP-resistant cells. METHODS: Human lung adenocarcinoma A549 (parental cells) and A549/DDP cells (DDP-resistant cells) were treated with different concentrations of DDP, the drug resistance index (RI) was determined by CCK-8 assay and the autophagy associated proteins, like Beclin 1, LC3Ⅱ and p62 were measured by Western blot. A549 and A549/DDP cells were treated with 10 μmol/L DDP (about IC50 of A549 cells), the cell viability was determined by CCK-8 assay, the autophagy and apoptosis associated proteins (including Beclin 1, LC3Ⅱ, p62, Bcl-2, Bax and cleaved-caspase 3) were measured by Western blot, and the activity of transcription factor FOXO3a and its subcellular localization were detected by Western blot and laser confocal scanning microscopy. Finally, the autophagy inhibitor Baflomycin A1 (Baf A1) and the protein kinase inhibitor of PI3K/Akt signaling pathway were co-treated with DDP respectively to test the mechanism of drug resistance. RESULTS: Compared with the parental A549 cells, the acquired resistant A549/DDP cells showed DDP-resistance and higher level of basal autophagy. More survival count of A549/DDP cells than that of A549 cells in the same environment stress of 10 μmol/L DDP. 10 μmol/L DDP induced A549 cells apoptosis by down-regulated Bcl-2, and increased Bax and cleaved-caspase 3, which followed the inhibition of PI3K/Akt signaling pathway and up-regulated the expression level of transcription factor FOXO3a. While the same concentration of DDP activated A549/DDP cells autophagy by up-regulated Beclin 1 and LC3Ⅱ, down-regulated p62, which followed the inhibition of PI3K/Akt signaling pathway and inhibited the phosphorylation of FOXO3a. CONCLUSION: DDP induces apoptosis by up-regulating the transcription factor FOXO3a via inhibiting the PI3K/Akt/FOXO3a signaling pathway in A549 cells, while activating autophagy by inducing the phosphorylation of FOXO3a via inhibiting the PI3K/Akt/FOXO3a signaling pathway in A549/DDP cells.
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OBJECTIVE@#To observe the effects of electroacupuncture (EA) on reproductive outcomes in women with Shen (Kidndy) deficiency syndrome after in vitro fertilization-embryo transfer (IVF-ET), and explore the underlying molecular mechanism.@*METHODS@#Sixty-six infertile patients with Shen deficiency syndrome undergoing IVF-ET were divided into EA or control groups according to a random table, 33 cases in each group. Before undergoing IVF, patients in the EA and control groups received EA therapy and placebo needle puncture, respectively, for 3 menstrual cycles. Shen deficiency syndrome scores were assessed. Other outcome measures included the number of retrieved oocytes and fertilization, high-quality embryo and clinical pregnancy rates. Follicular fluid was collected on the day of oocyte retrieval, and granulosa cell expression of phosphatidylinositide 3-kinases (PI3K), serine-threonine kinase (Akt) and forkhead box O3 (Foxo3a) mRNA were measured by reverse transcribed and quantitative real-time polymerase chain reaction.@*RESULTS@#Syndrome scores for pre- versus post-treatments decreased significantly (16.53±1.75 to 8.67±1.61) in the EA group (P<0.05), but showed no significant change in the control group (17.18±1.58 to 14.74±1.58). A significant difference in score change was found between the EA and control groups (P<0.05). High-quality embryo and clinical pregnancy rates were both increased in the EA group compared with the control group [69.15% (195/282) vs. 60.27% (176/292) and 66.67% (22/33) vs. 42.42% (14/33), respectively, P<0.05]. The fertilization rate was equivalent in EA and control groups. No difference was found in the number of retrieved oocytes between the two groups. Granulosa cell expression levels of PI3K and Akt mRNA were significantly increased in the EA group compared with the control group, while the expression of Foxo3a was reduced (all P<0.05).@*CONCLUSIONS@#For infertile patients with Shen deficiency syndrome undergoing IVF, EA for tonifying Shen as an adjunct treatment may alleviate clinical symptoms and improve the high-quality embryo rate. The EA-induced mechanism may involve regulation of PI3K/Akt/Foxo3a expression in granulosa cells to improve the developmental microenvironment of oocytes and inhibit granulosa cell apoptosis, possibly contributing to the improved clinical pregnancy rate (Registration No. ChiCTR 1800016217).