Lupeol Drug Alters The Microrna Expression Profiles In HELA Cells

As a natural plant extract, Lupeol is recently proved to be an effective drug to suppress cancer cells grow. To reveal its inner anti-cancer molecular mechanisms will be helpful on solving problems on drug resistance of anti-cancer medicines as well as on promoting the research process of new style of drug combinations. On the other hand, the field of anti-cancer is focusing on researches about miRNA, with its synthesis mechanism almost clear and ways of regulation increase rapidly. What's more, more and more miRNA's function explained by researchers make softwares for high-throughput prediction of microRNA targets and secondary structure analysis based on sequence analyzing increasingly sophisticated. With hela cells, this research build a model of Lupeol and Cisplatin and the analyze of the data is based on changes in the expression amount of miRNA.Using U6 as an internal reference, the research uses miRNA chip to deal with the model and gives it a full miRNA analyze. Through compares of the results of the miRNA chip, we found out that the expression amount of several kinds of miRNA is very similar and showing a cluster distribution in Bitmap. Meanwhile, target gene of the same cluster of miRNA, such as ERBB4, TEAD and MeCP2, is closely related to the regulation of cancer cells.In later studies, I analyzed the result of miRNA chip by significant difference and cluster analysis. Acompanied with current researches, I found that the expression changes of miR-504 and miR-34a are probably related with P53, which is a very important anti-cancer gene. On this basis, we performed real-time PCR to investigate the effects with different treatment time, the results accorded with the miRNA chip analysis, which confirmed our findings that miR-504 demonstrates a significant lower expression while miR-34a shows minor changes from 6h to 24h.According to targetscan and the current research findings, P53 is the direct target gene for miR-504.There are two binding spots of miR-504 seed on the 3'UTR sequence of P53, so miR504 can directly repress the expression of P53. In reference to the anti-cancer effect of lupeol, we think miR-504 and P53 are the two potential factors of lupeol to inhibit the growth of cancer cells. On the other side, BCL-2,E2F3,E2F5,CDK4,CDK6,Cyclin E2 and c-myc etc. are all proved to be the target genes of miR-34, and these proteins are all involved in P53's regulation of cell apoptosis and cell cycle, which is believed to be related with the canceration of cells. Thus, we conclude that "miR-504-p53-miR-34a-protien-regulating factors" are the molecular paths of lupeol to inhibit the growth of cancer cells. From the results of our research, the upregulation of miR-34a by P53 might occur after 24h of medical treatment.