Antiproliferative Effects Of Cardamonin Mediated By Anti-inflammation Via MTOR On Ovarian Cancer Cells

Object:Inflammation is closely related to the growth, invasion and angiogenesis of ovariancancer, and anti-inflammation is expected to become a new strategy for ovarian cancertherapy. Cardamomin (CAR) exhibits a variety of pharmacological activities includinganti-inflammatory and anti-tumor. The effects of CAR were associated with theinhibition on NF-κB and mTOR. However, whether the anti-inflammatory effect ofCAR is mediated by mTOR needs to be illustrated. This study was to investigate theproliferation inhibitory effect of CAR based on anti-inflammation and its possible target,which provides experimental evidence for the development of this kind of drug.Methods1Cell lineHuman ovarian cancer cell line SKOV32Grouping and Drug DeliveryLipopolysaccharide(LPS) and Pyrrolidine dithiocarbamate(PDTC) were adopt toinduce inflammation and block the NF-κB mediated inflammatory pathwaysrespectively. The control group, LPS pretreated group and PDTC+LPS pretreated groupwere setted for experiments; and then the cells were treated with RAP and differentdoses of CAR. The concentrations of different drugs were as follow: LPS （1μg/mL）,PDTC（100μM）、RAP（0.1μM）、CAR（100μM、30μM、10μM、3μM、1μM、0.1μM）.3Measurement and methodology3.1Cellular morphology and density were observed under the microscope.3.2Cell proliferation was measured by MTT method. 3.3The mRNA expression of inflammatory factor Interleukin-6(IL-6) was detectedby semi-quantitative PCR.3.4The protein expressions of NF-κB p65, mTOR, p-mTOR, S6K1, p-S6K1proteinwere detected by Western Blot.Result1. The proliferation of the normal and LPS pretreated SKOV3cells was inhibited byCAR in a dose-dependent manner.2. The mRNA expression of inflammatory factor IL-6and protein expression ofnucleus NF-κB p65were increased by LPS stimulation (P<0.01). The expressionof IL-6and nucleus NF-κB p65was inhibited by CAR in both kinds cells. In thenormal cells, the inhibitory effect of low dose CAR on IL-6mRNA expression wasstronger than that of RAP and high dose CAR (P<0.01); however, in the LPSpretreated cells, the inhibitory effect of high dose CAR and RAP was stronger thanthat of low dose CAR.3. Treated with LPS, mTOR and S6K1were activated and the expression of p-mTORand p-S6K1were increased significantly (P<0.01). The phosphorylation of mTORand S6K1was inhibited by CAR in both kinds cells (P<0.01), and the inhibitoryeffect of high dose CAR was stronger. CAR had no significant effect on the totalprotein expression of mTOR and S6K1.4. The expression of LPS induced nuclear NF-κB p65was inhibited significantly byPDTC (P<0.01), and PDTC had no significant effect on the expression of p-mTORand p-S6K1. Compared with LPS pretreated group, the inhibitory effect of RAPand high dose CAR in PDTC+LPS pretreated group on the expression of p-mTORand p-S6K1was weaker. Pretreated with PDTC+LPS, RAP and high dose CARhad no significant effect on the expression of nuclear NF-κB p65.ConclusionCAR inhibits the cell proliferation of the normal group and SKOV3induced by LPS,its mechanism may be related to that CAR reduces the phosphorylation of mTOR andS6K1and thus affect the expression of NF-κB and the IL-6.