Intervention Study Of Allicin On Cell Biological Behavior Of HeLa Cells In Cervical Carcinoma

Objective: To study the efficacy of allicin on human cervicalcarcinoma HeLa cells in celluar proliferation and apoptosis and exploredthe possible molecular regulation mechanism.Methods: The WST-8assay was used to detect the OD values and thegrowth inhibition rates of HeLa cells at24-72h which cultured withallicin in different concentrations(2.5ug/ml,5ug/ml,10ug/ml,20ug/ml,40ug/ml,80ug/ml,160ug/ml).By flow cytometry(FCM),Apoptosis andcell cycle distribution in HeLa cells were measured at24h after treatmentwith different concentrations allicin(2.5ug/ml,5ug/ml,10ug/ml,20ug/ml,40ug/ml,80ug/ml,160ug/ml)and at24-72h after10ug/ml allicin. After24h treatment of different concentration allicin(20ug/ml,40ug/ml,80ug/ml),the total cellular RNA and total cellular protein wereextracted.The mRNA expression levels of β-catenin,c-myc and cyclinD1were tested by RT-PCR.The neucleoprotein level of β-catenin was testedby Westem-blot. All the experiments were performed in triplicates.Statistical differences between groups were calculated using SPSS17.0.Differences were considered statistically significant for values P <0.05.Result:After24-72h treatment with2.5-160ug/ml allicin,the cellproliferation inhibition rate was increased from24%to82%in a dose andtime-dependent manner by WST-1assay.The difference was statistically significant (P<0.05).which indicated that allicin significantly inhibitedHeLa proliferation in a dose and time-dependent manner.The apoptosis cell rate was rised from (8.54±1.53)%to (63.06±5.32)%when HeLa cells were treated with different concentrationsallicin(2.5ug/ml,5ug/ml,10ug/ml,20ug/ml,40ug/ml,80ug/ml,160ug/ml)for24h. After cultured with10ug/ml allicin for24-72h, the rate wascounted from (9.37±1.39)%to (61.84±3.74)%. The difference wasstatistically significant (P<0.05). It suggested that allicin could induceqpoptosis of HeLa cells.which was clearly dose and time-dependent.After treatment with allicin for24h, there was not a significantlydifference in cell-cycle distribution between each group when theconcentration of allicin is more than or equal to10ug/ml. However,whenthe dose was more than10ug/ml,the percentage of cells in G0/G1phasewas increased from (38.2±1.62)%to(66.6±3.25)%while the percentageof cells in S phase was dncreased from (43.1±1.02)%to (24.8±1.19)%.The difference was statistically significant (P<0.05). After treatment with10ug/ml allicin for24-72h, the number of cells in G0/G1phase wasincreased from (30.10±1.62)%to(47.2±4.24)%,while the number ofcells in S phase was dncreased from (53.1±1.02)%to (35.2±2.35)%,The difference was statistically significant (P<0.05). Which demonstratedthat allicin could induce cell-cycle arrest in G1/S phase.With increasing drug concentrations (20μg/ml,40μg/ml,80μg/ml), the expression of nucleoprotein β-cateninthe and the mRNA levels ofβ-catenin,c-myc and cyclinD1were significantly downregulated after24htreatment with allicin. The difference was statistically significant(P<0.05).It illustrated that Allicin could down-regulate the mRNAexpression levels of c-myc and cyclinDl and promote the degradation ofnucleoprotein β-catenin.Which may inhibite the proliferation andpromote the apoptosis of HeLa cells.Conclusion: By down-regulated the expression levels of nucleoproteinβ-catenin, allicin could inhibite the activation of Wnt signalingpathways,down-regulated the mRNA expression levels of c-myc andcyclinDl, prevent the activation of Rb-E2F pathway and inducecell-cycle arrest in G1/S phase to inhibite the proliferation and promotethe apoptosis of human cervical carcinoma HeLa cells.