Effect On Mirnas And Its Targets Of Hela And Hela/DDP By Beta-sitosterol

β-sitosterol is a kind of plant sterols which has little toxic and side effects, highsafety and rich resources. It has been widely paid attention by people because of itsanti-cancer ability.It has already become a people’s top priority to study its tumorsuppressor mechanism and then to applied it to cancer prevention and treatment bett-er.MicroRNAs are of great significance in tumor study.It has been the focus of resear-ch to inhibit tumorigenesis through regulate its expression. We analysis the differenti-ally expressed of miRNAs and its target genes, predict the possible pathway of tumorsuppression through the beta sitosterol treatment of Hela and Hela/DDP cell,and aim atfinding the early molecular markers of tumorigenesis, and the the basis of Drugscreening.We use different concentration of β-sitosterol（50μmol/L,37.5μmol/L,25μmol/L,12.5μmol/L,6.25μmol/L）to treat Hela and Hela/DDP cells for24h. The results showthatβ-sitosterol can inhibit cell proliferation of Hela and Hela/DDP, and we found thatthe inhibitory effect of drug to Hela cells is more obvious than to Hela/DDP cells.We use microRNAs gene chip technology and bioinformatics tools to make highthroughput screening of744microRNAs and forecast the targets of Hela cell afterbeta-sitosterol24h treatment by TargetScan Basement and the analysis of minimumfree energy(MFE).We find that hsa-miR-21, hsa-miR-143, hsa-miR-504, hsa-miR-10bare contribute to tumorigenesis and also provide direct evidence that PDCD4may be thetarget of hsa-miR-21, RAC3and TP53may be the targets of hsa-miR-504, TP53maybe the targets of hsa-miR-143and TIAM1may be the targets of hsa-miR-10b.We use β-sitosterol（25μmol/L）to deal with two kinds of cells, and choose3h,6h,12h,18h,24h as time node.After that we perform quantitative analysis of miRNAsand its target genes through the method of real-time quantitative PCR. The resultsshowed that:(1) In Hela cells, β-sitosterol treatment down-regulates the quantity ofhsa-miR-21, hsa-miR-504, upregulates the quantity of hsa-miR-10b and makeshsa-miR-143increase first and reduced subsequently. Otherwise the target genes TP53,PDCD4are upregulated, RAC3is downregulated and JNK1is upregulated first anddownregulatd subsequently.(2) In Hela/DDP, β-sitosterol treatment down-regulates thequantity of hsa-miR-21， upregulates the quantity of hsa-miR-10b and hsa-miR-143.There is no fluorescent signal of hsa-miR-504during Drug treatment process.Otherwise the target genes TP53, PDCD4,JNK1are upregulated and RAC3isdownregulated. These results suggest thatβ-sitosterol may induce the apoptosis of Helaand Hela/DDP through regulate the quantity of microRNAs and the expression of itetargets TP53、PDCD4、RAC3、JNK1.