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国家自然科学基金(81271050)

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相关作者:王宜强王宜强更多>>
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The involvement of proline-rich protein Mus musculus predicted gene 4736 in ocular surface functions
2016年
AIM: To research the two homologous predicted proline -rich protein genes, Mus musculus predicted gene 4736 (MP4) and proline-rich protein BstNI subfamily 1 (Prb1) which were significantly upregulated in cultured corneal organs when encountering fungal pathogen preparations. This study was to confirm the expression and potential functions of these two genes in ocular surface. METHODS: A Pseudomonas aeruginosa keratitis model was established in Balb/c mice. One day post infection, mRNA level of MP4 was measured using real-time polymerase chain reaction (PCR), and MP4 protein detected by immunohistochemistry (IHC) or Western blot using a customized polyclonal anti -MP4 antibody preparation. Lacrimal glands from normal mice were also subjected to IHC staining for MP4. An online bioinformatics program, BioGPS, was utilized to screen public data to determine other potential locations of MP4. RESULTS: One day after keratitis induction, MP4 was upregulated in the corneas at both mRNA level as measured using real -time PCR and protein levels as measured using Western blot and IHC. BioGPS analysis of public data suggested that the MP4 gene was most abundantly expressed in the lacrimal glands, and IHC revealed that normal murine lacrimal glands were positive for MP4 staining. CONCLUSION: MP4 and Prb1 are closely related with the physiology and pathological processes of the ocular surface. Considering the significance of ocular surface abnormalities like dry eye, we propose that MP4 and Prb1 contribute to homeostasis of ocular surface, and deserve more extensive functional and disease correlation studies.
Xia QiSheng-Wei RenFeng ZhangYi-Qiang Wang
小鼠基质重塑相关7(MXRA7)基因产物在肝内互作蛋白质的鉴定(英文)被引量:3
2019年
基质重塑相关7(matrix remodeling associated 7,MXRA7)基因于2002年被命名,但无论在人类或其他动物体系,该基因或其蛋白质产物的功能均未知。直至我们最近的研究表明,该基因可能参与眼睛发育或肝损伤及修复。在本研究中,应用酵母双杂交策略,用小鼠MXRA7诱饵载体对小鼠肝cDNA文库进行筛选,发现23种蛋白质(MUP1, Cpt1a, Mat1a, aldh1l1, Cytb, H2-K1, Psmb1, marc2, Atp5j2, Sec24D, Trf, Rdh7, Apoe, Glud1, Gmfb, Alb, Hdlbp, Pzp, Etnk2, Nrn1, Serpina1a, Apoa2, GNMT)可能直接或间接地与MXRA7蛋白发生相互作用。其中,主要尿蛋白1(major urinary protein,MUP1)或其同家族蛋白质占所有候选克隆的1/6,提示其很可能是小鼠肝内与MXRA7蛋白有较强相互作用的蛋白质。通过基因重组在Hepa 1-6肝癌细胞系中同时过表达MXRA7和MUP1蛋白,荧光染色证明这两种蛋白质在细胞内共定位,应用抗MXRA7或MUP1的抗体进行Pull-down检测,则证明二者共同存在于细胞裂解液中。另外,重组MXRA7蛋白和MUP1蛋白在无任何其他蛋白质的缓冲液体系中直接形成复合体。因此,至少在小鼠肝组织体系中,MXRA7蛋白可能通过与MUP1等蛋白质的相互作用而发挥作用。
王梦汝沈莹李园园王宜强
关键词:酵母双杂交
基质重塑相关7(MXRA7)对SHI-1白血病细胞系生物学功能的影响被引量:1
2022年
目的:构建过表达人全长基质重塑相关7(MXRA7)的SHI-1急性髓系白血病稳转细胞系,检测MXRA7对SHI-1细胞生物学功能的影响。方法:合成人MXRA7全长的cDNA序列,并连接到慢病毒穿梭载体pRRL-Venus中,使用293T细胞包装产生慢病毒,感染人白血病细胞系SHI-1,应用流式细胞术分选YFP;细胞,扩大培养获得稳转细胞系。通过Real-time qPCR、Western blot及激光共聚焦技术验证MXRA7在SHI-1细胞中的表达及分布。采用流式细胞术检测细胞增殖和细胞周期,采用Annexin V和7-AAD染色流式检测细胞凋亡,采用Western blot技术检测与细胞凋亡相关蛋白的表达情况。结果:成功构建了过表达MXRA7的SHI-1稳转细胞系,激光共聚焦证明MXRA7表达于SHI-1细胞的胞浆中;与对照细胞相比,过表达MXRA7不会影响细胞增殖和细胞周期,但会降低甲氨蝶呤诱导的凋亡细胞的百分比,促进凋亡抑制蛋白BCL-2的表达。结论:成功构建了过表达MXRA7的SHI-1稳转细胞系,MXRA7通过促进BCL-2的表达来抑制药物诱导的细胞凋亡。
郑宇丹孙振江马鲲鹏王宜强林丹丹
关键词:急性髓系白血病细胞凋亡BCL-2
基质重塑相关7(MXRA7)基因:一个非亲缘家族的一员
2020年
本世纪初,有学者对1375份人类cDNA文库内的所有基因进行共表达分析,发现有8个转录本总是与基质重塑相关的基因(如胶原、基质金属蛋白酶、骨成型蛋白等)共表达,将它们命名为matrixremodeling associated 1~8(MXRA1~8,暂译“基质重塑相关1~8”基因),但它们彼此之间并无序列同源性。随后的研究确定了该家族中除MXRA7外其他7个成员的生物学功能,其中部分成员在研究中以其他别名出现,比如MXRA4即CD93或C1qRP。国内学者首先在关于角膜新生血管和角膜感染的研究中报道MXRA7表达的变化,继而在化学诱导的肝损伤模型、免疫介导的银屑病模型以及骨髓间充质干细胞向成骨细胞、成脂细胞分化模型中证实MXRA7的重要作用。基因组文库数据显示MXRA7在脊椎动物中保守存在,且在人和小鼠等模型生物中均可以产生多种蛋白异构体。大量公共数据库(比如GEO)中都包含MXRA7在不同实验体系中的表达水平和变化信息,众多基于高通量分析的文献也常在数据流水账中对MXRA7一笔带过。对这些资源进行汇总分析,提示MXRA7基因产物可分布于胞外基质、胞浆中甚至细胞核内,可参与胚胎发育、细胞分化、细胞代谢等多种生理过程,或参与神经系统、心血管系统、骨骼系统的病理过程,或参与肿瘤、白血病、免疫性疾病、感染等疾病的发病机制。本文对现有文献和公共数据进行综述,以期激发学界对MXRA7的关注。
王宜强
MXRA7 is involved in megakaryocyte differentiation and platelet production
2023年
Matrix remodeling is a critical process in hematopoiesis.The biology of MXRA7,as a matrix remodeling associated gene,has still not been reported in hematopoietic process.Public databases showed that MXRA7 expressed in hematopoietic stem cells,suggesting that it may be involved in hematopoiesis.We found that the amounts of megakaryocytes were lower in bone marrow and spleen from Mxra7−/−mice compared with that from wild-type mice.Knock-out of MXRA7 also reduced the amount of platelet in peripheral blood and affected the function of platelets.Knock-out of MXRA7 inhibited hematopoietic stem/progenitor cells differentiate to megakaryocytes possibly through down-regulating the expression of GATA-1 and FOG-1.Moreover,knockdown of MXRA7 in MEG-01 cells could inhibit the cell proliferation and cell apoptosis.Knockdown of MXRA7 inhibited the differentiation of MEG-01 cells and proplatelet formation through suppressing the ERK/MAPK signaling pathway and the expression ofβ-tubulin.In conclusion,the current study demonstrated the potential significance of MXRA7 in megakaryocyte differentiation and platelet production.The novel findings proposed a new target for the treatment of platelet-related diseases,and much more investigations are guaranteed to dissect the mechanisms of MXRA7 in megakaryocyte differentiation and platelet production.
Zhenjiang SunBenfang WangYing ShenKunpeng MaTing WangYiqiang WangDandan Lin
关键词:DIFFERENTIATIONMEGAKARYOCYTESPLATELET
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