The Effect Of Triptolide On Epigenetics In T-cell Acute Lymphoblastic Leukemia

Objective: To study whether TPL has effect on the Wnt signaling pathway in T-cell acute lymphoblastic leukemia(T-ALL) and the possible mechanisms under which.Methods: 1. The cell viability of T-ALL cell lines jurkat and molt4 before and after TPL treatment was detected by Cell Counting Kit-8 assay.2. The expressions of b-catenin, C-MYC, DNMT1 and DNMT3 A before and after TPL treatment at protein level were detected by western blot.3. Detected expressions of WIF1, SOX17, CDH1, SFRP5, C-MYC, TCF7, DNMT1 and DNMT3 A before and after TPL treatment at transcriptional level by semi-quantitative RT-PCR.4. Tested methylation status of WIF1, SOX17, CDH1 and SFRP5 by using Methylation specific polymerase chain reaction(MSP).Results: 1. After treatment by TPL, cell viability of jurkat and molt4 was decreased in a significant dose- and time-dependent manner.2. Expressions of b-catenin, C-MYC, DNMT1 and DNMT3 A at protein level were downregulated after TPL treatment.3. Expressions of WIF1, SOX17, CDH1 and SFRP5 at transcriptional level were increased after TPL treatment, while expressions of C-MYC, TCF7, DNMT1 and DNMT3 A were decreased after TPL treatment.4. After TPL treatment, methylation status of WIF1, SFRP5 and CDH1 were changed and unmethylated gene were upregulated. While methylation status of SOX17 did not change obviously.Conclusions: Results of our study indicated that inhibition of TPL to T-ALL cell lines may be in part through inhibiting to Wnt signaling pathway by upregulating Wnt antagonists. Upregulation of SFRP5, CDH1 and WIF1 may result from changes of methylation status. But methylation status of SOX17 did not change apparently, which indicated that other mechanism may also be involved.