• 생명과학회지
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• 제27권10호
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• pp.1104-1110
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• 2017

히스톤 H3K4의 메칠화는 3T3-L1의 지방세포의 분화를 촉진하는 것으로 알려져 있으나, 히스톤 H3K4 메칠화 효소인 SET1A가 지방세포 분화를 조절하는지에 대해서는 보고된 바가 없다. 그러므로 본 연구에서는 SET1A의 3T3-L1 지방세포의 분화조절과 기전을 연구하였다. SET1A의 발현은 3T3-L1 지방세포 분화과정에서 증가함을 관찰하였다. 3T3-L1 지방전구세포에서 siRNA을 이용하여 SET1A의 발현을 감소시키면 3T3-L1 지방전구세포의 분화가 억제됨을 관찰하여 SET1A가 3T3-L1 지방전구세포의 분화를 촉진함을 알 수 있었다. 이에 대한 조절기전을 알기 위해, SET1A의 발현을 감소시킨 3T3-L1 지방전구세포의 세포증식을 측정한 결과, 분화 초기 단계인 분화 후 2일 동안 3T3-L1 지방세포의 증식이 감소하였다. 또한 분화 후 7일 동안 지방세포세포 분화 조절인자들의 발현을 측정한 결과, SET1A의 발현을 감소시킨 3T3-L1 지방세포에서 $PPAR{\gamma}$의 발현이 감소하였다. 위와 같은 연구결과를 바탕으로, SET1A는 분화초기단계에서는 mitotic clonal expansion 단계를 촉진하고, 분화후기단계에서는 $PPAR{\gamma}$의 발현을 증가시켜 3T3-L1 지방세포의 분화를 촉진함을 알 수 있었다.

• Reproductive and Developmental Biology
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• 제37권4호
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• pp.269-279
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• 2013

Low efficiency of somatic cell nuclear transfer (SCNT) is attributed to incomplete reprogramming of transfered nuclei into oocytes. Trichostatin A (TSA), histone deacetylase inhibitor and 5-aza-2'deoxycytidine (5-aza-dC), DNA methylation inhibitor has been used to enhance nuclear reprogramming following SCNT. However, it was not known molecular mechanism by which TSA and 5-aza-dC improve preimplantation embryo and fetal development following SCNT. The present study investigates embryo viability and gene expression of cloned porcine preimplantation embryos in the presence and absence of TSA and 5-aza-dC as compared to embryos produced by parthenogenetic activation. Our results indicated that TSA treatment significantly improved development. However 5-aza-dC did not improve development. Presence of TSA and 5-aza-dC significantly improved total cell number, and also decreased the apoptotic and autophagic index. Three apoptotic-related genes, Bak, Bcl-xL, and Caspase 3 (Casp3), and three autophagic-related genes, ATG6, ATG8, and lysosomal-associated membrane protein 2 (LAMP2), were measured by real time RT-PCR. TSA and 5-aza-dC treatment resulted in high expression of anti-apoptotic gene Bcl-xL and low pro-apoptotic gene Bak expression compared to untreated NT embryos or parthenotes. Furthermore, LC3 protein expression was lower in NT-TSA and NT-5-aza-dC embryos than those of NT and parthenotes. In addition, TSA and 5-aza-dC treated embryos displayed a global acetylated histone H3 at lysine 9 and methylated DNA H3 at lysine 9 profile similar to the parthenogenetic blastocysts. Finally, we determined that several DNA methyltransferase genes Dnmt1, Dnmt3a and Dnmt3b. NT blastocysts showed higher levels Dnmt1 than those of the TSA and 5-aza-dC blastocysts. Dnmt3a is lower in 5-aza-dC than NT, NTTSA and parthenotes. However, Dnmt3b is higher in 5-aza-dC than NT and NTTSA. These results suggest that TSA and 5-aza-dC positively regulates nuclear reprogramming which result in modulation of apoptosis and autophagy related gene expression and then reduce apoptosis and autophagy. In addition, TSA and 5-aza-dC affects the acetylated and methylated status of the H3K9.

• Molecules and Cells
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• 제45권4호
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• pp.202-215
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• 2022

The androgen receptor (AR) is an important therapeutic target for treating prostate cancer (PCa). Moreover, there is an increasing need for understanding the AR-independent progression of tumor cells such as neuroendocrine prostate cancer (NEPC). Menin, which is encoded by multiple endocrine neoplasia type 1 (MEN1), serves as a direct link between AR and the mixed-lineage leukemia (MLL) complex in PCa development by activating AR target genes through histone H3 lysine 4 methylation. Although menin is a critical component of AR signaling, its tumorigenic role in AR-independent PCa cells remains unknown. Here, we compared the role of menin in AR-positive and AR-negative PCa cells via RNAi-mediated or pharmacological inhibition of menin. We demonstrated that menin was involved in tumor cell growth and metastasis in PCa cells with low or deficient levels of AR. The inhibition of menin significantly diminished the growth of PCa cells and induced apoptosis, regardless of the presence of AR. Additionally, transcriptome analysis showed that the expression of many metastasis-associated genes was perturbed by menin inhibition in AR-negative DU145 cells. Furthermore, wound-healing assay results showed that menin promoted cell migration in AR-independent cellular contexts. Overall, these findings suggest a critical function of menin in tumorigenesis and provide a rationale for drug development against menin toward targeting high-risk metastatic PCa, especially those independent of AR.