Prediction Of MiRNA Related Antitumor Pathways Of Lupeol Based On High-throughput Sequencing Analysis

As a kind of natural products, triterpenes have been found to have a variety of biological activities. Lupeol, a triterpene found in various plants, is proved to involve in different signal pathways in the aspect of antitumor process, including improving cancer cell apoptosis, arresting cell cycle, inducing differentiation, drug sensitization, inhibiting tumorgenesis, etc. Lupeol could also regulate miRNAs, which act as pioneer factor in antitumor pathways, but the regulation signals have not yet been revealed. This research aims at studying genome wide expression changes in HeLa cell lines under the treatment of lupeol from three levels, miRNA, transciptome, and proteome, with the method of high throughtput quantification techniques, and exploring the molecular pathways of HeLa cell apoptosis induced by lupeol, based on the results of real time quantitiative PCR.On account of the calculation by GFOLD and DESeq, we selected out9significantly upregulated miRNAs, including hsa-miR-1307-3p, hsa-miR-193b-3p, hsa-miR-140-3p, hsa-miR-940, hsa-miR-889-3p, hsa-miR-1307-5p, hsa-miR-1293, hsa-miR-3124-5p, hsa-miR-152-5p, and6significantly downregulated miRNAs, consisting of hsa-miR-152-3p, hsa-miR-550a-3p, hsa-miR-183-5p, hsa-miR-27a-3p, hsa-miR-182-5p, hsa-miR-99b-3p, in HeLa cell lines after the12h treatment of100?M lupeol. Based on the regulation relationship between miRNA and its targets, and the analysis between predicted miRNA targets by Diana-microT-CDS and miRanda and dysregulated transcriptome and proteome, we filtered out hsa-miR-183-96-182cluster, KIAA1199, and PBX1for relative expression level verification with real time quantatitive PCR, under gradient medication and different time node. We found out that KIAA1199is the target of hsa-miR-183, and PBX1is the target of hsa-miR-182, in which hsa-miR-96might also participate.In accordance with published results, we further deducted the downstream pathways, in which there are PGRMC1, BAX, AKAP95, FADD, Wnt, Caspase family, HOX, ER?, and MEOX1, and conclude that lupeol takes part in tumor inhibition by suppressing tumor cell proliferation, promoting apoptosis, restaining migration, and sensitizing cancer cells after chemotherapy.