Salicylic Acid Activates AMPK/TSC2 To Down Regulate MTOR’s Growth Promotion Effect In Human Ovarian Cancer Cells

Background and Objective:Ovarian cancer is the most common malignant tumor of female genital tract.Due to the characteristics of ovarian cancer,the prognosis of patients is often poor and the recurrence rate is high,it’s a serious threat to women’s health.Current research confirms that mammalian target of rapamycin（mTOR）is highly expressed in ovarian cancer tissues.Abnormal activation of mTOR plays an important role in the occurrence and development of malignant tumor,mTOR has the function of anti apoptosis and promoting the growth of cancer cells.However,AMPK can activate Tuberous Sclerosis Complex 2（TSC2）directly by phosphorylation of threonine on1345 sites of TSC2,to formation of TSC1/TSC2 complex,final to inhibiting m TOR activity.Some research shows,salicylic acid can directly activate AMPK/TSC2,to down regulation of mTOR expression and activation,final to inhibit the growth of cancer cells.Therefore,we propose that salicylic acid can inhibit the growth of human ovarian cancer cells.The objective of this study is to investigate the effects of salicylic acid on human ovarian cancer A2780 and SKOV-3 cells by studying the expression and activation of proteins include AMPK、mTOR and TSC2,in order to provide a new way of thinking for the treatment of ovarian cancer.Methods:1、The inhibitory effect of salicylic acid on the growth of human ovarian cancer（A2780andSKOV-3cells）were investigated by using crystal violet assay;2、The inhibitory effect of cisplatin/paclitaxel on the growth of human ovarian cancer（A2780andSKOV-3cells）by using crystal violet assay.And to compare the survival rate of salicylic acid combined with cisplatin/paclitaxel and cisplatin/paclitaxel alone in human ovarian cancer cells（A2780andSKOV-3cells）;3、Western blot assay was used to detect the expression and activation of AMPK、mTOR and TSC2 in human ovarian cancer A2780 and SKOV-3 cells induced by salicylic acid(IC₂₀/IC₅₀)or cisplatin(IC₂₀/IC₅₀)or salicylic acid combined with cisplatin(SA-IC₂₀+DDP-IC₅₀)for 24 hours.Results:1、Salicylic acid could inhibit the growth of human ovarian cancer A2780 and SKOV-3 cells in a dose-dependent manner.And the 50%inhibitory concentration of salicylic acid in A2780 and SKOV-3 cells after 72 hours were:（3.870±0.270） mmol/L and（3.03±0.240）mmol/L;2、The 50%inhibitory concentration of paclitaxel/cisplatin in A2780 and SKOV-3cells after 72 hours respectively:（6.980±0.538）μmol/L and（6.138±0.440）μmol/L,（0.018±0.001）μmol/L and（0.023±0.002）μmol/L.After the combined of salicylic acid（IC20）,the IC50 of paclitaxel and cisplatin on human ovarian cancerA2780 and SKOV-3 cells were decreased to:（0.008±0.001）μmol/L and（0.01±0.001）μmol/L,（3.180±0.190）μmol/L and（2.690±0.160）μmol/L,the Dose Enhance Factor respectively:0.44 and 0.43,0.46 and 0.44..The results showed that compared with paclitaxel or cisplatin alone,the inhibition of cell growth was more obvious after the combined action of salicylic acid（P<0.05）,suggesting that cisplatin or paclitaxel combined with salicylic acid was superior to cisplatin or paclitaxel alone;3、Salicylic acid can increase the level of AMPK and TSC2 phosphorylation in human ovarian cancer A2780 and SKOV-3 cells,and significantly down regulate the expression of p-m TOR（P<0.05）.Conclusion:1、Salicylic acid can synergistic increase the inhibitory effect of paclitaxel or cisplatin on human ovarian cancer cells;2、Salicylic acid could upregulate the ovarian cancer cells AMPK and TSC2 phosphorylation,thereby inhibiting the expression of p-mTOR,may provide a new direction for the improvement of clinical chemistry of ovarian cancer treatment.