Stathmin 1(stmn1)gene,which is involved in the control of cell cycle progression,cell proliferation,differentiation and motility,is a crucial relay in the signal transduction of multiple signaling pathways.Reports on ...Stathmin 1(stmn1)gene,which is involved in the control of cell cycle progression,cell proliferation,differentiation and motility,is a crucial relay in the signal transduction of multiple signaling pathways.Reports on the function of stmn1 in fish are scarce.In this study,the characterization and function of stmn1a gene were explored in an important food fish,namely bighead carp(Hypophthalmichthys nobilis).The bighead carp stmn1a(Hynstmn1a)genomic sequence,which has 2995 bp with an ORF of 447 bp,is evolutionarily conserved in teleost fish.The highest level of expression of Hynstmn1a is in the un-fertilized eggs and spleen of adult bighead carp.The overexpression of Hynstmn1a facilitates cell cycle progression and suppresses apoptosis in EPC cells.In an siRNA assay,disruption of stmn1a in EPC cells resulted in the inhibition of cell cycle progression and promotion of apoptosis.Analysis of promoter activity indicated that an upstream transcription factor(foxo4)regulates Hynstmn1a.Zebrafish mutants with stmn1a knockout by CRISPR/Cas9 exhibited a significant retardation of body length(p<0.05).There was no significant difference in body weight between stmn1a-mutant zebrafish and wildtype(p=0.053).A 22-bp Indel in the promoter region of Hynstmn1a with an observed heterozygosity(Ho)of 0.136 in a bighead carp population(n=191).The polymorphism of this 22-bp Indel genotype is significantly associated with body length(BL),head length(HL)(p<0.05),body weight(BW),head height(HH),and head width(HW)(p<0.01),showing its genetic effects on growth promotion.This study sheds light on the function of stmn1a in fish and demonstrates the potential of the 22-bp Indel of Hynstmn1a as a gene marker when selecting breeding individuals for faster growth and a bigger head of bighead carp in aquaculture.展开更多
目的研究微管解聚蛋白1(Stathmin-1,STMN1)受特化蛋白1(specialized protein 1,SP1)转录调控对子宫平滑肌瘤细胞增殖和凋亡的作用。方法收集2017年6月至2019年6月在南京同仁医院妇产科接受子宫全切除和次全切除的30例子宫肌瘤患者的肌...目的研究微管解聚蛋白1(Stathmin-1,STMN1)受特化蛋白1(specialized protein 1,SP1)转录调控对子宫平滑肌瘤细胞增殖和凋亡的作用。方法收集2017年6月至2019年6月在南京同仁医院妇产科接受子宫全切除和次全切除的30例子宫肌瘤患者的肌瘤组织及周围(瘤旁)平滑肌,RT-qPCR检测子宫平滑肌瘤组织和周围平滑肌组织中STMN1和SP1 mRNA水平。CCK-8、TUNEL和Western blot检测敲低STMN1后人子宫平滑肌瘤(human uterine leiomyoma,HuLM)细胞增殖和凋亡。利用JASPAR网站预测转录因子SP1和STMN1启动子的结合位点。采用RT-qPCR、Western blot、荧光素酶、ChIP验证SP1和STMN1的转录调控,进行功能拯救实验。结果相较于正常平滑肌细胞,子宫平滑肌瘤组织中STMN1和SP1表达上调(P<0.001)。敲低STMN1抑制HuLM细胞增殖并促进凋亡(P<0.001)。SP1作为转录因子正向调控STMN1表达水平,敲低SP1能够抑制HuLM细胞增殖并促进凋亡(P<0.001),进一步过表达STMN1逆转敲低SP1对细胞增殖和凋亡的作用(P<0.001)。结论转录因子SP1通过上调STMN1促进子宫平滑肌瘤细胞增殖并抑制凋亡。展开更多
基金supported by the grants from National Natural Science Foundation of China(32072973)Special Fund for Strategic Pilot Technology of the Chinese Academy of Sciences(XDA24030505).
文摘Stathmin 1(stmn1)gene,which is involved in the control of cell cycle progression,cell proliferation,differentiation and motility,is a crucial relay in the signal transduction of multiple signaling pathways.Reports on the function of stmn1 in fish are scarce.In this study,the characterization and function of stmn1a gene were explored in an important food fish,namely bighead carp(Hypophthalmichthys nobilis).The bighead carp stmn1a(Hynstmn1a)genomic sequence,which has 2995 bp with an ORF of 447 bp,is evolutionarily conserved in teleost fish.The highest level of expression of Hynstmn1a is in the un-fertilized eggs and spleen of adult bighead carp.The overexpression of Hynstmn1a facilitates cell cycle progression and suppresses apoptosis in EPC cells.In an siRNA assay,disruption of stmn1a in EPC cells resulted in the inhibition of cell cycle progression and promotion of apoptosis.Analysis of promoter activity indicated that an upstream transcription factor(foxo4)regulates Hynstmn1a.Zebrafish mutants with stmn1a knockout by CRISPR/Cas9 exhibited a significant retardation of body length(p<0.05).There was no significant difference in body weight between stmn1a-mutant zebrafish and wildtype(p=0.053).A 22-bp Indel in the promoter region of Hynstmn1a with an observed heterozygosity(Ho)of 0.136 in a bighead carp population(n=191).The polymorphism of this 22-bp Indel genotype is significantly associated with body length(BL),head length(HL)(p<0.05),body weight(BW),head height(HH),and head width(HW)(p<0.01),showing its genetic effects on growth promotion.This study sheds light on the function of stmn1a in fish and demonstrates the potential of the 22-bp Indel of Hynstmn1a as a gene marker when selecting breeding individuals for faster growth and a bigger head of bighead carp in aquaculture.
