目的:探讨长链非编码RNA TFAP2A-AS1对子宫内膜癌细胞增殖、侵袭和迁移的影响及机制。方法:选择50例子宫内膜癌组织和对应的癌旁组织。选取子宫内膜癌细胞株(RL95-2、HEC-1A、HHUA、HEC-1B及Ishikawa),子宫内膜上皮细胞hEEC。子宫内膜...目的:探讨长链非编码RNA TFAP2A-AS1对子宫内膜癌细胞增殖、侵袭和迁移的影响及机制。方法:选择50例子宫内膜癌组织和对应的癌旁组织。选取子宫内膜癌细胞株(RL95-2、HEC-1A、HHUA、HEC-1B及Ishikawa),子宫内膜上皮细胞hEEC。子宫内膜癌细胞株Ishikawa转染si-TFAP2A-AS1(si-TFAP2A-AS1组)、si-NC(si-NC组)、miR-9-5p mimics(miR-9-5p组)及miR-NC(miR-NC组);RL95-2细胞转染OE-TFAP2A-AS1(TFAP2A-AS1组)、Vector(Vector组)、sh-miR-9-5p(sh-miR-9-5p组)及sh-NC(sh-NC组)。CCK-8检测细胞增殖能力,Transwell检测细胞侵袭、迁移能力,双荧光素酶实验、pull down实验分析TFAP2A-AS1与miR-9-5p的靶向关系;miR-9-5p与ERK的靶向关系,实时荧光定量PCR(RT-qPCR)检测子宫内膜癌组织、癌旁组织、子宫内膜癌细胞及hEEC细胞TFAP2AAS1、miR-9-5p表达水平,Western blot检测细胞外调节蛋白激酶(ERK)、磷酸化ERK(p-ERK)水平。结果:子宫内膜癌组织中TFAP2A-AS1表达水平低于癌旁组织,miR-9-5p表达水平高于癌旁组织(P<0.05)。子宫内膜癌组织TFAP2A-AS1、miR-9-5p表达水平与分化程度、肿瘤直径、TNM分期及淋巴结转移有关。子宫内膜癌组织中TFAP2A-AS1、miR-9-5p表达水平呈负相关(r=-0.782,P=0.002)。与hEEC细胞比较,RL95-2、HEC-1A、HHUA、HEC-1B及Ishikawa细胞TFAP2A-AS1表达水平降低,miR-9-5p表达水平升高(P<0.05)。与si-NC组比较,si-TFAP2A-AS1组细胞48 h、72 h OD值,侵袭、迁移细胞数均升高(P<0.05);与Vector组比较,TFAP2A-AS1组细胞48 h、72 h OD值,侵袭、迁移细胞数均降低(P<0.05)。与miR-NC组比较,miR-9-5p组细胞48 h、72 h OD值,侵袭、迁移细胞数均升高(P<0.05);与sh-NC组比较,sh-miR-9-5p组细胞48 h、72 h OD值,侵袭、迁移细胞数均降低(P<0.05)。TFAP2A-AS1负性调控miR-9-5p、p-ERK表达水平,miR-9-5p可靶向调控ERK蛋白。结论:过表达TFAP2A-AS1通过靶向下调miR-9-5p,抑制ERK通路,最终抑制子宫内膜癌细胞恶性生物学行为。展开更多
In vitro, mouse embryonic stem(ES) cells can differentiate into many somatic cell types, including neurons and glial cells. When cultured in serum-free medium, ES cells convert spontaneously and efficiently to a neura...In vitro, mouse embryonic stem(ES) cells can differentiate into many somatic cell types, including neurons and glial cells. When cultured in serum-free medium, ES cells convert spontaneously and efficiently to a neural fate. Previous studies have shown that the neural conversion of mouse ES cells includes both the participation of neural-specific transcription factors and the regulation of epigenetic modifications. However, the intracellular mechanism underlying this intrinsic transition still remains to be further elucidated. Herein, we describe a long intergenic non-coding RNA, Linc RNA1230, which participates in the regulation of the neural lineage specification of mouse ES cells. The ectopic forced expression of Linc RNA1230 dramatically inhibited mouse ES cells from adopting a neural cell fate, while Linc RNA1230 knockdown promoted the conversion of mouse ES cells towards neural progenitors. Mechanistic studies have shown that Linc RNA1230 inhibits the activation of early neural genes, such as Pax6 and Sox1, through the modulation of bivalent modifications(tri-methylation of histone3 lysine4 and histone3 lysine27) at the promoters of these genes. The interaction of Linc RNA1230 with Wdr5 blocked the localization of Wdr5 at the promoters of early neural genes, thereby inhibiting the enrichment of H3K4me3 modifications at these loci. Collectively, these findings revealed a crucial role for Linc RNA1230 in the regulation of the neural differentiation of mouse ES cells.展开更多
背景:目前非编码RNA已被证实在多种骨科疾病中发挥重要作用,因此,非编码RNA可能为了解该疾病的病因、发病机制、治疗方法带来新的思路。目的:探索早期诊断、治疗股骨头坏死的方法和思路,综述非编码RNA在股骨头坏死中的研究进展,并提出...背景:目前非编码RNA已被证实在多种骨科疾病中发挥重要作用,因此,非编码RNA可能为了解该疾病的病因、发病机制、治疗方法带来新的思路。目的:探索早期诊断、治疗股骨头坏死的方法和思路,综述非编码RNA在股骨头坏死中的研究进展,并提出目前所存在的问题及解决方法。方法:以“股骨头坏死,非编码RNA,miRNA,lncRNA,circRNA”为检索词检索中国知网、万方、维普等数据库,以“osteonecrosis of the femoral head,non-coding RNAs,miRNA,lncRNA,circRNA”为检索词检索PubMed、Medline等数据库,检索时限均为2000年1月至2021年11月。最终纳入67篇文献。结果与结论:(1)非编码RNA的种类非常丰富,大约占所有转录产物的98%,同时占所有RNA的90%左右,根据其功能可分为管家非编码RNA(如tRNA、rRNA、snoRNA)以及调节非编码RNA(如miRNA、lncRNA、circRNA、siRNA);(2)目前miRNAs与激素型股骨头坏死的研究逐渐成为热点,而外泌体lncRNAs、circRNAs与股骨头坏死的相关研究尚未见到报道;(3)多数非编码RNA在发生激素型股骨头坏死时表达下调,大部分非编码RNA通过负向调控其相关靶基因,以促进骨髓间充质干细胞增殖、成骨分化,促进成骨标记物的表达,也可以促进血管内皮细胞生成,从而抑制股骨头缺血性坏死的发生及进展;(4)外泌体携带的非编码RNA也可以在股骨头坏死中发生作用,但目前文献量较少,非编码RNA是否通过外泌体携带所产生的作用目前暂不清楚,在未来的研究中可逐渐完善;(5)利用非编码RNA类似物和抑制剂可以有效抑制非创伤性股骨头坏死的发生发展,可以为非创伤性股骨头坏死的早期诊断和治疗提供新的策略。展开更多
文摘目的:探讨长链非编码RNA TFAP2A-AS1对子宫内膜癌细胞增殖、侵袭和迁移的影响及机制。方法:选择50例子宫内膜癌组织和对应的癌旁组织。选取子宫内膜癌细胞株(RL95-2、HEC-1A、HHUA、HEC-1B及Ishikawa),子宫内膜上皮细胞hEEC。子宫内膜癌细胞株Ishikawa转染si-TFAP2A-AS1(si-TFAP2A-AS1组)、si-NC(si-NC组)、miR-9-5p mimics(miR-9-5p组)及miR-NC(miR-NC组);RL95-2细胞转染OE-TFAP2A-AS1(TFAP2A-AS1组)、Vector(Vector组)、sh-miR-9-5p(sh-miR-9-5p组)及sh-NC(sh-NC组)。CCK-8检测细胞增殖能力,Transwell检测细胞侵袭、迁移能力,双荧光素酶实验、pull down实验分析TFAP2A-AS1与miR-9-5p的靶向关系;miR-9-5p与ERK的靶向关系,实时荧光定量PCR(RT-qPCR)检测子宫内膜癌组织、癌旁组织、子宫内膜癌细胞及hEEC细胞TFAP2AAS1、miR-9-5p表达水平,Western blot检测细胞外调节蛋白激酶(ERK)、磷酸化ERK(p-ERK)水平。结果:子宫内膜癌组织中TFAP2A-AS1表达水平低于癌旁组织,miR-9-5p表达水平高于癌旁组织(P<0.05)。子宫内膜癌组织TFAP2A-AS1、miR-9-5p表达水平与分化程度、肿瘤直径、TNM分期及淋巴结转移有关。子宫内膜癌组织中TFAP2A-AS1、miR-9-5p表达水平呈负相关(r=-0.782,P=0.002)。与hEEC细胞比较,RL95-2、HEC-1A、HHUA、HEC-1B及Ishikawa细胞TFAP2A-AS1表达水平降低,miR-9-5p表达水平升高(P<0.05)。与si-NC组比较,si-TFAP2A-AS1组细胞48 h、72 h OD值,侵袭、迁移细胞数均升高(P<0.05);与Vector组比较,TFAP2A-AS1组细胞48 h、72 h OD值,侵袭、迁移细胞数均降低(P<0.05)。与miR-NC组比较,miR-9-5p组细胞48 h、72 h OD值,侵袭、迁移细胞数均升高(P<0.05);与sh-NC组比较,sh-miR-9-5p组细胞48 h、72 h OD值,侵袭、迁移细胞数均降低(P<0.05)。TFAP2A-AS1负性调控miR-9-5p、p-ERK表达水平,miR-9-5p可靶向调控ERK蛋白。结论:过表达TFAP2A-AS1通过靶向下调miR-9-5p,抑制ERK通路,最终抑制子宫内膜癌细胞恶性生物学行为。
基金supported by National Natural Science Foundation of China (81530042, 31571529, 31210103905, 31571519, 31571390, 31371510, 31301208, 31471250, 31401257)the Ministry of Science and Technology (2012CB966603, 2013CB967600, 2013CB967401)+2 种基金Science and Technology Commission of Shanghai Municipality (15JC1403200, 15JC1403201)Shanghai Rising-Star Program (14QA1403900)the Fundamental Research Funds for the Central Universities (2000219099)
文摘In vitro, mouse embryonic stem(ES) cells can differentiate into many somatic cell types, including neurons and glial cells. When cultured in serum-free medium, ES cells convert spontaneously and efficiently to a neural fate. Previous studies have shown that the neural conversion of mouse ES cells includes both the participation of neural-specific transcription factors and the regulation of epigenetic modifications. However, the intracellular mechanism underlying this intrinsic transition still remains to be further elucidated. Herein, we describe a long intergenic non-coding RNA, Linc RNA1230, which participates in the regulation of the neural lineage specification of mouse ES cells. The ectopic forced expression of Linc RNA1230 dramatically inhibited mouse ES cells from adopting a neural cell fate, while Linc RNA1230 knockdown promoted the conversion of mouse ES cells towards neural progenitors. Mechanistic studies have shown that Linc RNA1230 inhibits the activation of early neural genes, such as Pax6 and Sox1, through the modulation of bivalent modifications(tri-methylation of histone3 lysine4 and histone3 lysine27) at the promoters of these genes. The interaction of Linc RNA1230 with Wdr5 blocked the localization of Wdr5 at the promoters of early neural genes, thereby inhibiting the enrichment of H3K4me3 modifications at these loci. Collectively, these findings revealed a crucial role for Linc RNA1230 in the regulation of the neural differentiation of mouse ES cells.
文摘背景:目前非编码RNA已被证实在多种骨科疾病中发挥重要作用,因此,非编码RNA可能为了解该疾病的病因、发病机制、治疗方法带来新的思路。目的:探索早期诊断、治疗股骨头坏死的方法和思路,综述非编码RNA在股骨头坏死中的研究进展,并提出目前所存在的问题及解决方法。方法:以“股骨头坏死,非编码RNA,miRNA,lncRNA,circRNA”为检索词检索中国知网、万方、维普等数据库,以“osteonecrosis of the femoral head,non-coding RNAs,miRNA,lncRNA,circRNA”为检索词检索PubMed、Medline等数据库,检索时限均为2000年1月至2021年11月。最终纳入67篇文献。结果与结论:(1)非编码RNA的种类非常丰富,大约占所有转录产物的98%,同时占所有RNA的90%左右,根据其功能可分为管家非编码RNA(如tRNA、rRNA、snoRNA)以及调节非编码RNA(如miRNA、lncRNA、circRNA、siRNA);(2)目前miRNAs与激素型股骨头坏死的研究逐渐成为热点,而外泌体lncRNAs、circRNAs与股骨头坏死的相关研究尚未见到报道;(3)多数非编码RNA在发生激素型股骨头坏死时表达下调,大部分非编码RNA通过负向调控其相关靶基因,以促进骨髓间充质干细胞增殖、成骨分化,促进成骨标记物的表达,也可以促进血管内皮细胞生成,从而抑制股骨头缺血性坏死的发生及进展;(4)外泌体携带的非编码RNA也可以在股骨头坏死中发生作用,但目前文献量较少,非编码RNA是否通过外泌体携带所产生的作用目前暂不清楚,在未来的研究中可逐渐完善;(5)利用非编码RNA类似物和抑制剂可以有效抑制非创伤性股骨头坏死的发生发展,可以为非创伤性股骨头坏死的早期诊断和治疗提供新的策略。
