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中国生物化学与分子生物学会2019年全国学术会议暨学会...

作者： Qian Qian Cai Ying Yang Xing Zhong Wu Department of Biochemistry and Molecular Biology School of Basic Medical Sciences Fudan University Key Lab of Glycoconjugate Research Ministry of Public Health

Metal responsive transcription factor 1(MTF1) is a zinc dependent transcription factor which is involved in the regulation of intracellular signaling pathways. We noted that the expression and half-life of MTF1 mRNA were increased after sulfatide treatment, and elevated MTF1 expression was significantly associated with low survival rate in HCC patients. We also found that sulfatide inhibited the N6-methyladenosine(m6A) methylation of MTF1 mRNA by MeRIP-seq. As the m6A modification is reversible due to the interplay of methyltransferase(MTase) ‘writer’ and demethylase ‘eraser’ enzymes, with the methyl group was recognized by m6A ‘reader’ proteins. We further detected whether sulfatide influenced the expressions of m6A binding proteins(YTHDF1, YTHDF2, YTHDF3), the m6A methyltransferases(METTL3, METTL14 and WTAP) and the m6A demethylases(ALKBH5 and FTO). Interestingly, even though sulfatide made no difference in the m6A ‘writers,erasers or reader’ expression, but inhibited the METTL3-METTL14-WTAP complex interaction. In conclusion, this study demonstrated that sulfatide may regulated MTF1 m6A modification by interfering the formation of m6A writer complex.

关键词： MTF1 sulfatide HCC m6A methylation m6A writer complex

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sulfatide Interaction with BRD1 Promotes Integrin αV Gene Expression

Sulfatide Interaction with BRD1 Promotes Integrin αV Gene E...

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中国生物化学与分子生物学会2016年全国学术会议

作者： QIAN QIAN CAI YI WEI DONG BING QI XIAO TING SHAO RONG WANG ZHONG YI CHEN BAO MEI HE HE YANG XU XING ZHONG WU Department of Biochemistry and Molecular Biology Shanghai Medical College Fudan University Key Lab of Glycoconjugate Research Ministry of Public Health Yu Ying Hospital Wenzhou Medical University

IntegrinαV is important in promotion of anchorage independence,angiogenesis and cancer metastasis,but its regulation is not well understood especially in hepatocellular carcinoma(HCC).Here we show that high integrinαV expression was closely associated with poor prognosis of patients with HCC and sulfatide activated integrinαV transcription through

关键词： bromodomain containing protein 1 sulfatide histone acetylation

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SIN3B promotes integrin αV subunit gene transcription and cell migration of hepatocellular carcinoma

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Journal of Molecular Cell Biology 2019年第5期11卷 421-432页

作者： Qianqian Cai Yuanyuan Liu Ping Zhu Chunlang Kang Heyang Xu Bing Qi Rong Wang Yiwei Dong Xing Zhong Wu Department of Biochemistry and Molecular Biology School of Basic Medical Sciences Fudan University. Key Lab of Glycoconjugate Research Ministry of Public Health Shanghai 200032 China Zhejiang Provincial People's Hospital Hangzhou 310053 China

Paired amphipathic helix protein (SIN3B) is a transcription corepressor for many genes. Here we show a different regulation mechanism of integrin aV gene expression by SIN3B in human hepatocellular carcinoma (HCC). We first observed a close relationship between Integrin aV and SIN3B expressions in HCC patients and tumor cell lines with different metastatic potentials. Overexpression of SIN3B significantly accelerated the cell migration rate of SMMC-7721, but failed when integrin αV expression was silenced. Interestingly, SIN3B stimulated integrin aV subunit promoter activity only in the presence of sulfatide. Importantly, SIN3B was identified in the complex with sulfatide by mass spectrometry. Fat blot assay indicated that SIN3B specifically interacted with sulfatide. Molecular modeling suggested that sulfatide induced the conformational change of SIN3B from compacted α-helices to a relaxed β-sheet in PAH2 domain. The data of immunoprecipitation and ChIP assay indicated that altered SIN3B lost the binding affinity with MAD1 and HDAC2, which reduced the recruitment of HDAC2 on integrin aV gene promoter and prevented the deacetylation of the histone 3. In conclusion, this study demonstrated that SIN3B promoted the transcriptional activation of the integrin αV subunit gene promoter by reducing interaction with HDAC2.

关键词： paired amphipathic helix protein ITGAV sulfatide HCC histone deacetylation

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Therapeutic Effects of Myriocin in Experimental Alcohol-Related Neurobehavioral Dysfunction and Frontal Lobe White Matter Biochemical Pathology

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Journal of Behavioral and Brain Science 2022年第2期12卷 23-42页

作者： Camilla Homans Emine B. Yalcin Ming Tong Gina Gallucci David Bautista Natalia Moriel Suzanne de la Monte Biotechnology Graduate Program Brown University Providence RI USA Department of Pathology and Laboratory Medicine Warren Alpert Medical School of Brown University Providence RI USA Liver Research Center Department of Medicine Rhode Island Hospital Providence RI USA Warren Alpert Medical School of Brown University Providence RI USA Brown University Providence RI USA Department of Pathology and Laboratory Medicine Rhode Island Hospital Women and Infants Hospital of Rhode Island Providence VA Medical Center Providence RI USA

Background & Objective: Chronic excessive alcohol consumption causes white matter degeneration with myelin loss and impaired neuronal conductivity. Subsequent rarefaction of myelin accounts for the sustained deficits in cognition, learning, and memory. Correspondingly, chronic heavy or repeated binge alcohol exposures in humans and experimental models alter myelin lipid composition leading to build-up of ceramides which can be neurotoxic and broadly inhibitory to brain functions. Methods: This study examined the effects of chronic + binge alcohol exposures (8 weeks) and intervention with myriocin, a ceramide inhibitor, on neurobehavioral functions (Open Field, Novel Object Recognition, and Morris Water Maze tests) and frontal lobe white matter myelin lipid biochemical pathology in an adult Long-Evans rat model. Results: The ethanol-exposed group had significant deficits in executive functions with increased indices of anxiety and impairments in spatial learning acquisition. Myriocin partially remediated these effects of ethanol while not impacting behavior in the control group. Ethanol-fed rats had significantly smaller brains with broadly reduced expression of sulfatides and reduced expression of two of the three sphingomyelins detected in frontal white matter. Myriocin partially resolved these effects corresponding with improvements in neurobehavioral function. Conclusion: Therapeutic strategies that support cerebral white matter myelin expression of sulfatide and sphingomyelin may help remediate cognitive-behavioral dysfunction following chronic heavy alcohol consumption in humans.

关键词： Adolescence Alcohol Behavior Binge Drinking Brain Atrophy Myriocin Neurodegeneration Rat sulfatide White Matter

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人体外周血中Ⅱ型NKT细胞比例及功能初探

人体外周血中Ⅱ型NKT细胞比例及功能初探

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第十二届全国免疫学学术大会

作者：刘宇梁智辉翁秀芳吴雄文华中科技大学同济医学院

背景:CD1d限制性T细胞根据其TCR组成可分为TCR组成相对恒定的Ⅰ型NKT细胞与TCR组成多样的Ⅱ型NKT细胞。不同于Ⅰ型NKT细胞在小鼠内富集,Ⅱ型NKT细胞在人体内比例较高,并在自身免疫性疾病,肿瘤及感染免疫中发挥重要的作用。然而现有研究... 详细信息

背景:CD1d限制性T细胞根据其TCR组成可分为TCR组成相对恒定的Ⅰ型NKT细胞与TCR组成多样的Ⅱ型NKT细胞。不同于Ⅰ型NKT细胞在小鼠内富集,Ⅱ型NKT细胞在人体内比例较高,并在自身免疫性疾病,肿瘤及感染免疫中发挥重要的作用。然而现有研究对Ⅱ型NKT细胞认知极少,其在人体内明确的比例与功能尚属未知,极大地限制了Ⅱ型NKT细胞相关免疫疾病认识与干预手段建立。方法:本实验将Ⅱ型NKT细胞特异性抗原sulfatide加载的K562-CD1d细胞(sulfatide-K562-CD1d)作为刺激细胞,并设立Ⅰ型NKT细胞特异性抗原α-GalCer加载的K562-CD1d细胞(αGalCer-K562-CD1d)作为对照,对比分析正常人群外周血中PBMC接受刺激后细胞因子分泌情况;并制备sulfatide/CD1d四聚体检测人外周血中sulfatide特异性Ⅱ型NKT细胞比例。结果:我们发现:对比于αGalCer-K562-CD1d细胞,sulfatide-K562-CD1d细胞能够刺激外周血PBMC细胞产生更多的IFN-γ/IL-2/IL-6/IL-10/IL-17/TNF-α等细胞因子,提示人体外周血中含有比Ⅰ型NKT细胞更高比例的Ⅱ型NKT细胞。通过sulfatide加载的CD1d四聚体与空载的CD1d四聚体对外周血中PBMC染色分析发现,sulfatide反应性Ⅱ型NKT细胞占外周血PBMC比例较低,仅为0.1%左右,然而外周血中非sulfatide反应性Ⅱ型NKT细胞的比例有待后续试验进一步检测。讨论:本研究对健康个体外周血中sulfatide反应性的Ⅱ型NKT细胞的比例与功能进行了初步研究。在后续实验中,需要进一步应用酶联免疫斑点(ELISPOT)法明确其它脂类抗原反应性Ⅱ型NKT细胞的比例,并对正常人群及不同疾病患者中Ⅱ型NKT细胞的数量与功能进行对比分析,以探索Ⅱ型NKT细胞在免疫相关疾病中的作用及可能干预靶点。

关键词： Ⅱ型NKT sulfatide CD1d四聚体

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