The gonad specific expression of insulin-like growth factor 3(igf3)has suggested an important role of igf3 in fish reproduction.In this study,medaka igf3 was isolated and its expression patterns were compared with igf...The gonad specific expression of insulin-like growth factor 3(igf3)has suggested an important role of igf3 in fish reproduction.In this study,medaka igf3 was isolated and its expression patterns were compared with igf1,and germ cell gene vasa in adult medaka ovary.Molecular cloning and sequencing showed that the open reading frame(ORF)of medaka igf3 was comprised of 501 nucleotides and encoded 166 amino acid residues.Reverse transcription polymerase chain reaction(RT-PCR)analysis showed that igf3 RNA was specifically expressed in developing embryos and adult gonads of both sexes.Real time quantitative PCR(q-PCR)indicated that igf3 expression was gradually increased during oogenesis,and reached its highest level at stage V.Using Chromogenic and fluorescent in situ hybridization,igf3 was shown to be present in the germ cells of oogonia and in oocytes at stages I-III,as well as abundant in granulosa cells and theca cells of oocytes at stages IV-V.In addition,igf3 and igf1 were expressed mutually in the outer theca cells of stage V oocytes in ovary.Collectively,we demonstrated that gonad specific igf3 could mark medaka ovarian somatic cells and germ cells.These findings highlight the importance of igf3 during ovarian development.展开更多
基金This work was supported by the National Natural Science Foundation of China(31672700)National Key R&D Program of China(2018YFD0901205).
文摘The gonad specific expression of insulin-like growth factor 3(igf3)has suggested an important role of igf3 in fish reproduction.In this study,medaka igf3 was isolated and its expression patterns were compared with igf1,and germ cell gene vasa in adult medaka ovary.Molecular cloning and sequencing showed that the open reading frame(ORF)of medaka igf3 was comprised of 501 nucleotides and encoded 166 amino acid residues.Reverse transcription polymerase chain reaction(RT-PCR)analysis showed that igf3 RNA was specifically expressed in developing embryos and adult gonads of both sexes.Real time quantitative PCR(q-PCR)indicated that igf3 expression was gradually increased during oogenesis,and reached its highest level at stage V.Using Chromogenic and fluorescent in situ hybridization,igf3 was shown to be present in the germ cells of oogonia and in oocytes at stages I-III,as well as abundant in granulosa cells and theca cells of oocytes at stages IV-V.In addition,igf3 and igf1 were expressed mutually in the outer theca cells of stage V oocytes in ovary.Collectively,we demonstrated that gonad specific igf3 could mark medaka ovarian somatic cells and germ cells.These findings highlight the importance of igf3 during ovarian development.
基金国家自然科学基金(82270162,82270224,82070178)北京市自然科学基金(7222175)+2 种基金军队卫勤保障能力创新与生成专项(21WQ034)保健专项科研课题重点项目(21BJZ30)国家重点研发计划(2021Y FA 1100904)。
文摘目的:探寻IGF2BP3基因表达水平与急性髓系白血病(AML)患者预后的关系。方法:通过对本中心27例AML患者骨髓原代白血病细胞进行转录组高通量测序,分析IGF2BP3基因表达水平与患者临床特征之间的关系,并在初治AML患者及难治AML(Refractory AML)患者样本中验证。分析20例健康对照者和26例AML患者中IGF2BP3基因表达水平的差异。采用RT-qPCR、Western blot检测两种蒽环类耐药细胞系(HL60/ADR、K562/ADR)中IGF2BP3表达水平,比较其与敏感细胞(HL60、K562)的表达差异。通过3个数据集,分析IGF2BP3在AML患者中的表达水平及与预后的关系,进一步使用Cox生存分析IGF2BP3在AML中的预后价值。结果:在本中心27例AML患者骨髓原代白血病细胞中,难治性AML患者的IGF2BP3表达量明显高于化疗敏感的患者(P=0.0343),白血病细胞髓外浸润(extramedullary infiltration,EMI)患者的IGF2BP3表达量明显高于无髓外浸润的AML患者(P=0.0049)。与健康人比较,IGF2BP3在AML患者中表达增加(P=0.0009)。蒽环类耐药细胞系(HL60/ADR、K562/ADR)中IGF2BP3 mRNA的表达显著高于敏感细胞系(K562/ADR vs K562,P=0.0430;HL60/ADR vs HL60,P=0.7369)。Western blot结果显示,耐药细胞中IGF2BP3蛋白表达显著高于敏感细胞(P<0.001)。qPCR结果显示,难治AML患者中IGF2BP3的mRNA表达水平明显高于化疗敏感患者(P=0.002)。在3个大样本AML患者队列中IGF2BP3高表达预示AML预后不良(P<0.05)。单因素和多因素预后分析证实IGF2BP3高表达与患者较短的无事件生存(HR=1.887,P=0.024)和总体生存(HR=1.619,P=0.016)显著相关。结论:IGF2BP3基因高表达可能是AML预后不良的重要因素,提示IGF2BP3基因有望成为AML的临床预后评估和提供治疗策略的新的分子标志物。
