TMED2 Induces Cisplatin Resistance in Breast Cancer via Targeting the KEAP1-Nrf2 Pathway

Objective Cisplatin is the first-line treatment for breast cancer,but it faces challenges of drug resistance.This study investigated new molecular mechanisms underlying cisplatin resistance in breast cancer.Methods We analyzed sequencing data from the TCGA database to identify potential associations between transmembrane emp24 protein transport domain containing 2(TMED2)and breast cancer.Western blotting,real-time PCR,CCK-8,and TUNEL assays were used to measure the effects and molecular mechanism of TMED2 on cisplatin resistance in MCF-7 and MDA-MB-231 cell lines.Results TMED2 was overexpressed in breast cancer and associated with poor prognosis.TMED2 increased cisplatin resistance in breast cancer cells in vitro via promoting ubiquitination of Kelch-like ECH-associated protein 1(KEAP1),relieving inhibition of KEAP1 on nuclear factor erythroid 2-related factor 2(Nrf2),and increasing expression of downstream drug resistance related genes,such as heme oxygenase 1(HO-1)and NAD(P)H quinone oxidoreductase 1(NQO1).Conclusion We identified a new molecular mechanism by which TMED2 affects cisplatin resistance in breast cancer.Our results provide theoretical guidance for future clinical applications.

All-trans Retinoic Acid Induced the Differentiation of Human Glioma Cells

OBJECTIVE To observe the effect of all-trans retinoic acid （ATRA） on inducing human glioma MO59K cells differentiation and further explore the underlying molecular mechanisms.METHODS The expression of glial fibrillary acidic protein （GFAP） was detected by immunocytochemistry staining. The mRNA levels of GFAP, retinoid X receptor α（RXRα）, p21 were examined by semi-quantitative RT-PCR analysis. Luciferase activity assay was performed in the COS-7, MO59K cells to measure p21 promoter transcription activity.RESULTS ATRA could significantly enhance the expression and mRNA level of GFAP by immunostaining and RT-PCR （P〈0.05）. Simultaneously, the mRNA levels of RXRα and p21 were remarkably increased in dose-dependent manner by RT-PCR （P〈0.05）. Furthermore, luciferase assay confirmed that ATRA and RXRα could transactivate p21 promoter in COS-7 and glioma cells （P〈0.05）.CONCLUSION ATRA can induce differentiation of human glioma cells. The RXRα and p21 were activated during ATRAinduced differentiation process. This effect may be caused by directly RXRα-induced p21 gene transactivation. Our findings provide novel evidence for the future studies to explore the molecular mechanism of transcriptional regulation for glioma cell differentiation and cellular therapeutic approaches for glioblastoma.

Transforming growth factor-β3 induced rat bone marrow-derived mesenchymal stem cells differentiation into smooth muscle cells by activating Myocardin

Bone marrow mesenchymal stem cells (MSCs) can differentiate into smooth muscle cells (SMCs) and have tremendous potential for cell therapy and tissue engineering. In this study, to understand the effects of TGF-β3 on rat bone marrow-derived MSCs and the underlying molecular mechanism of this differentiation process, we investigated that the changes of myocardin-related transcription factors (MRTFs) at the transcriptional level after rat MSCs were treated with TGF-β3. The results showed that TGF-β3 increased the expression of contractile genes, such as SM22, smooth muscle-myosin heavy chain (SM- MHC), SM-α-actin in MSCs. When TGF-β3 induced MSCs differentiation into SMCs, myocardin and MRTF-A were activated. The data indicated that TGF-β3 induced rat bone marrow-derived MSCs differentiation into SMCs by activating mypcardin and MRTF-A.

MRTF-A transactivates COMT gene and decreases the anti-tumor effects of tamoxifen

Myocardin-related transcription factors A (MRTF-A) is a myocardin-related transcription factor that have been found strongly activated CarG box–containing genes through its direct binding to serum response factor (SRF). In the present study, the MRTF-A ex-pression vector was constructed. The MTT assay showed that transfection of MRTF-A could significantly decrease the anti-tumor effect of tamoxifen on MCF-7 breast cancer cells. The bioinformatics analysis found that the CarG element existed in the pro-moter region of COMT gene of many familiar verte-brates, including of human, rhesus macaque, chimpanzee, etc. The results of RT-PCR assay further showed that MRTF-A could enhance the transcrip-tion level of COMT. These results are the first to indicate that COMT might be a target gene which could be regulated by MRTF-A/SRF, and such transactivation event might be involved in the process of tamoxifen resistance.