Curcumin Regulation Of Mirna Profile In Urine Bladder Cancer Cell T24 And The Associated Mechanism

Backgrounds and objectiveCurcumin is the major curcunoid of Indian spice turmeric. Its potential anticancer effects stem from its ability to inhibit cell growth and induce apoptosis in cancer cells. The mechanisms include interfering with multiple signaling pathway. Recent data show that curcumin can function through miRNA to combat tumor. miRNA is a short ribonucleic acid (RNA) molecules, on average only 22 nucleotides long. miRNAs are post-transcriptional regulators that bind to complementary sequences on target messenger RNA transcripts (mRNAs), usually resulting in translational repression and gene silencing. Curcumin can change miRNA profile in lung cancer and pancreatic cancer cells, and the disregulation of oncogenic miRNA were employed to function as antitumor pathway[1,2]. Curcumin induced differential expression profile are different in these two cell types. We investigate the differential expression profile induced by curcumin in urine trasitional cell carcinoma cell T24 to find the miRNA pathway involved in it's antitumor effection.MethodsAffemitrix miRNA microarray were used to detect the differential expression profile induced by curcumin in T24cells, the differential expression miRNA molecular screened by a threshold of two folds. miRPATH online software were used to mine the pathways which involved in differential expressed miRNA. Bioinformatics data of pathways were compared with literature reviews. Western blot assay were used to validate the target proteins of miRNAs involved these pathways.Results12.5μM curcumin treated T24 cells showed differential expression profile of miRNA (threshold is 2 folds). After 24h of curcumin treated, four miRNAs have been disregulated (hsa-miR-185 and hsa-miR-494 downregulated, hsa-miR-671-5p and hsa-miR-1307 upregulated). After 48h of curcumin treated, seven miRNAs have been disregulated (hsa-miR-181b, hsa-miR-138, hsa-miR-149, hsa-miR-339-5p, hsa-miR-193b, hsa-miR-455-3p and hsa-miR-744), and they are all upregulated.MirPATH online software (microT₄.0) mined 2946 target genes of differential expressed miRNAs. Among them 592 involved in KEGG signaling pathway. These pathways include six class of pathways named by KEGG. Among them singal transduction, cell mobility, cell growth and death, cell communication, cancer pathways are involved in oncogenesis and progression. We contrast bioinformatical outcome with literature and select several target proteins of differential expressed miRNAs to validate bioinformatical results. Western blot assay showed that cyclin D1 downregulated, proapoptosis factor Bax and Bid upregulated, nuclear transcription factor p65 downregulated, effector of TGF-βsingal cascade Smad3 and Smad4 downregulated, translation initiated factor EIF4E and VEGFA downregulated. ConclusionsCurcumin disreguated the miRNA expression profile in urine bladder carcinoma cell T24. The differential expressed miRNA target moleculars involved in signaling pathways may play an important role in curcumin's antitumor effections.