The Study On Apoptosis Of Ovarian Cancer SKOV-3/DDP Cells Induced By Allicin

Objective：To investigate P53, Bax and Bcl-2and other apoptosis-related gene expressionchanges of ovarian cancer cisplatin-resistant cell lines SKOV-3/DDP after treated byallicin, and explore the mechanism of induction of apoptosis of tumor cells.Methods：The SKOV-3/DDP cell were randomly divided into control group, allicingroup(40ug/ml), cisplatin group(40ug/ml) and allicin(40ug/ml)+cisplatinum(40ug/ml)group. Each group were cultured24h,48h. We used MTT method to calculateproliferation inhibition rate of each group.Then we used the Jin's formula to calculatethe two-drug combination index(q),and analyzed the effect of the two drugs incombination.E (AB) is the inhibition of the two drugs in combination,EA,EB is theinhibition rate of each drug used alone.when q=0.85~1.15, the performance of therole of two drugs is additive;q>1.15, the role of the two drugs in combination issynergistic;q<0.85, the role of two drugs is antagonistic.We used RT-PCR method toobserve of P53, Bax and Bcl-2mRNA expression of each group.Western blot wasused to observe P53, Bax and Bcl-2protein expression changes in each group.Result：1,The SKOV-3/DDP cell growth inhibition in allicin group, the cisplatin group,allicin+cisplatin platinum group was significantly increased,compared with thecontrol group (P<0.05). inhibition rate of the allicin+cisplatin group which culturedSKOV-3/DDP cell for24h was higher than its respective alone which cultured theSKOV-3/DDP cell (P <0.05),inhibition of the allicin+cisplatin group culturedSKOV-3/DDP cell for48h was higher than cisplatin group (P <0.05),But nostatistical significance between allicin group and the allicin+cisplatin group whichcultured SKOV-3/DDP cell for48h. The SKOV-3/DDP cell inhibition rate in Allicingroup, allicin+cisplatinum group was of time-dependent manner.The combinationindex of the combined treatment grous on SKOV-3/DDP cells was1.16after24h.After48h, the combination index was0.85.According to the Gin's formula,we can speculate, the performance of the role of two drugs was synergistic after24h; the roleof the two drugs in combination was additive after48h.2,Compared with the control group, the pro-apoptotic gene P53mRNA andprotein expression of cisplatin group was not significantly increased (P>0.05).Theexpression of pro-apoptotic gene P53mRNA and protein in allicin group, theallicin+cisplatin group was higher than cisplatin group and blank control group (P<0.05). The expression of P53mRNA and protein in allicin+cisplatin group was notstatistically significant between the expression of allicin group(P>0.05).3,The cisplatin group of pro-apoptotic gene Bax mRNA and protein expressionwas significantly increased,compared with the control group(P <0.05). Theexpression of pro-apoptotic gene Bax mRNA and protein in allicin group was higherthan cisplatin group(P<0.05).The expression of Bax mRNA and protein ofallicin+cisplatin group on SKOV-3/DDP cells was higher than their respective singleused group(P <0.05).4,Compared with the control group, the antiapoptotic genes Bcl-2mRNA andprotein expression of cisplatin group was not significantly increased (P>0.05). Theexpression of Bcl-2mRNA and protein in allicin group, the allicin+cisplatin groupwas lower than cisplatin group and blank control group (P <0.05). The expression ofBcl-2mRNA and protein in allicin+cisplatin group was lower than allicin group (P<0.05).Conclusion：1,Allicin Inhibited SKOV-3/DDP cell growth,Showing a time-effectrelationship. Allicin had the stronger inhibitory effect than cisplatin in SKOV-3/DDP.Allicin and cisplatin had synergistic/additive effect.2,Allicin increased the expression of pro-apoptotic gene P53,Bax mRNA andprotein, reduced expression of the anti-apoptotic gene Bcl-2mRNA and protein,which was superior to cisplatin. pro-apoptotic gene expression of P53,Bax mRNAand protein of Allicin combined with cisplatin was higher than the respective singlegroup,, and the anti-apoptotic gene Bcl-2mRNA and protein expression of Allicincombined with cisplatin was lower than the respective single group. 3,pro-apoptotic gene P53mRNA and protein expression levels in Allicin groupincreased,Along with the pro-apoptotic Bax mRNA and protein were significantlyhigher. Allicin induced apoptosis's mechanism may be induce P53mRNA andprotein expression firstly,then induce Bax mRNA and protein expression by the P53gene,played a chain of apoptosis activation effect.4,Bax mRNA and protein were significantly higher,With Bcl-2mRNA andprotein expression reduced in allicin group. allicin induced apoptosis's mechanismmay be through increasing Bax protein and down-regulating Bcl-2protein,andpromoting SKOV-3/DDP apoptosisIn short, allicin can inhibit SKOV-3/DDP cell growth and induce SKOV-3/DDPcell apoptosis,its mechanism may be related to upregulate expression of P53and Bax,and downregulate expression of Bcl-2.