Correlation Of Autophagy And AKT/mTOR Signaling Pathway In The Reversal Of Carboplatin Resistance In Ovarian Cancer Cells By Curcumin

Objective: This experiment was designed to investigate the mechanism of Akt-m TOR signaling pathway and Beclin1,LC3 and TRAP1 in reversing carboplatin resistance in curcumin.Methods: Cultured ovarian cancer transgenic cell line T2(constructed by Prof.Sandra Orsulic's group,p53 knockout rat ovarian epithelium,and ovarian cancer cell line that was transfected with c-myc and Akt)was detected by CCK-8 method.Curcumin,carboplatin,and the proliferation activity of the cells after the combination of the two drugs.The m RNA levels of Akt,m TOR,Beclin1,LC3 and TRAP1 in T2 cells after treatment with curcumin,carboplatin and two drugs were detected by Real-time PCR.Akt and Beclin1 were detected by Western blot after T2 cells were treated with T2 cells.,LC3,TRAP1 protein expression levels.The Akt protein level of T2 cells was knocked down and the transcription and protein expression levels of Beclin1,LC3 and TRAP1 were detected.The CCK-8 method was used to detect the sensitivity of T2 cells to curcumin,carboplatin,and the combination of two drugs after Knockdown of Akt protein.SPSS statistical software was used to analyze the results.Results: Curcumin and carboplatin all inhibited the proliferation of T2 cells.IC50 of curcumin was 21.2 ?mol/L at 48 hours and IC50 of carboplatin was 145.59 ?mol/L.Curcumin and carboplatin had synergistic effects.Curcumin and carboplatin reduced the expression of Akt and m TOR proteins in T2 cells,and the protein expression levels of Beclin1,LC3,and TRAP1 did not change significantly.When the two drugs were combined,the protein expression levels of LC3 and TRAP1 increased.After knockdown of Akt protein in T2 cells,the m RNA and protein expression levels of Beclin1,LC3,and TRAP1 in T2 cells increased.There was no significant change in the sensitivity of T2 cells to curcumin after knockdown of Akt protein,and the sensitivity to carboplatin was significantly increased.Low concentrations of curcumin can increase the sensitivity of T2 cells to carboplatin,and curcumin does not increase the sensitivity of T2 cell lines to carboplatin after Akt knockdown.Conclusion: Both curcumin and carboplatin can inhibit the expression of Akt,and the autophagy and the expression of mitochondrial autophagy-related proteins increase when the two drugs are combined.The combination of the two drugs may play a role in tumor cells by mediating the autophagic death of cells.Inhibition.The Akt-m TOR pathway has an inhibitory effect on autophagy in T2 cells.Knockdown of Akt T2 cells showed no significant change in the sensitivity of curcumin,indicating that Akt is not a key protein that curcumin inhibits T2 cells,and curcumin may exert inhibitory effects through other pathways.The sensitivity of Knockdown Akt T2 cells to carboplatin was significantly increased,indicating that reducing Akt protein levels can reverse the resistance of cells to carboplatin.Knockdown of curcumin in Akt T2 cells no longer improves the sensitivity of T2 cell lines to carboplatin,and Akt may be the key to curcumin's ability to reverse carboplatin resistance in ovarian cancer cells.