Background:Previously,phosphoinositide-3-kinase regulatory subunit 1(PIK3R1)and dual specificity phosphatase 1(DUSP1)were identified as promising candidate genes for milk production traits due to their being different...Background:Previously,phosphoinositide-3-kinase regulatory subunit 1(PIK3R1)and dual specificity phosphatase 1(DUSP1)were identified as promising candidate genes for milk production traits due to their being differentially expressed between the dry period and the peak of lactation in livers of dairy cows.Hence,in this study,the single nucleotide polymorphisms(SNPs)of PIK3R1 and DUSP1 genes were identified and their genetic associations with milk yield,fat yield,fat percentage,protein yield,and protein percentage,were investigated using 1067 Chinese Holstein cows from 40 sire families.Results:By re-sequencing the entire coding region and 2000 bp of the 5′and 3′flanking regions of the two genes,one SNP in the 5′untranslated region(UTR),three in the 3′UTR,and two in the 3′flanking region of PIK3R1 were identified,and one in the 5′flanking region,one in the 3′UTR,and two in the 3′flanking region of DUSP1 were found.Subsequent single-locus association analyses showed that five SNPs in PIK3 R1,rs42590258,rs210389799,rs208819656,rs41255622,rs133655926,and rs211408208,and four SNPs in DUSP1,rs207593520,rs208460068,rs209154772,and rs210000760,were significantly associated with milk,fat and protein yields in the first or second lactation(P values≤0.0001 and 0.0461).In addition,by the Haploview 4.2 software,the six and four SNPs in PIK3R1 and DUSP1 respectively formed one haplotype block,and the haplotype-based association analyses showed significant associations between their haplotype combinations and the milk traits in both two lactations(P values≤0.0001 and 0.0364).One SNP,rs207593520(T/G),was predicted to alter the transcription factor binding sites(TFBSs)in the 5′flanking region of DUSP1.Further,the dual-luciferase assay showed that the transcription activity of allele T in rs207593520 was significantly higher than that of allele G,suggesting the activation of transcriptional activity of DUSP1 gene by allele T of rs207593520.Thus,the rs207593520 SNP was highlighted as a potential causal mutation that should be further verified.Conclusions:We demonstrated novel and significant genetic effects of the PIK3R1 and DUSP1 genes on milk production traits in dairy cows,and our findings provide information for use in dairy cattle breeding.展开更多
目的探讨血浆外泌体源性miR-20a-5p通过靶向PIK3R1对雌激素受体阳性的乳腺癌(estrogen receptor-positive breast cancer,ER(+)BC)骨转移的影响。方法借助生物信息学网站检索与ER(+)BC骨转移相关的数据集,选择miR-20a-5p纳入研究。收集9...目的探讨血浆外泌体源性miR-20a-5p通过靶向PIK3R1对雌激素受体阳性的乳腺癌(estrogen receptor-positive breast cancer,ER(+)BC)骨转移的影响。方法借助生物信息学网站检索与ER(+)BC骨转移相关的数据集,选择miR-20a-5p纳入研究。收集90例ER(+)BC患者与相对应的健康人的血浆,检测血浆中PIK3R1的表达,从血浆中提取外泌体,检测外泌体中miR-20a-5p的表达。双荧光素酶报告实验验证miR-20a-5p与PIK3R1的靶向调控关系。分别将转染NC-inhibitor、miR-inhibitor的外泌体或转染si-NC、si-PIK3R1的ER(+)BC细胞注射到小鼠左心室,Micro-CT扫描骨组织并进行骨组织TRAP染色。将转染NC-inhibitor、miR-inhibitor的外泌体与转染si-NC、si-PIK3R1的ER(+)BC细胞共培养后,使用Western blot检测破骨细胞分子c-fos、NFATc1的表达。结果qRT-PCR检测发现较正常血浆外泌体(1±0.26)或细胞(1±0.13),miR-20a-5p在ER(+)BC血浆外泌体(1.49±0.27)(t=12.40,P<0.001)及BC细胞系MCF-7(1.64±0.13)(t=6.03,P=0.004)、BT474(1.49±0.11)(t=4.98,P=0.008)、T47D(1.98±0.15)(t=8.55,P=0.001)中均表达升高。而与正常血浆外泌体(1±0.25)或细胞(1±0.10)比较,PIK3R1在ER(+)BC血浆外泌体(0.69±0.24)(t=8.48,P<0.001)及ER(+)BC细胞系MCF-7(0.73±0.05)(t=4.18,P=0.014)、BT474(0.61±0.05)(t=6.04,P=0.004)、T47D(0.34±0.04)(t=10.61,P<0.001)中则表达降低。PIK3R1在血浆(t=8.48,P<0.001)及ER(+)BC细胞系MCF-7(t=4.18,P=0.014)、BT474(t=6.04,P=0.004)、T47D(t=10.61,P<0.001)中则表达降低。PIK3R1被证实为miR-20a-5p的靶基因。与NC-inhibitor组小鼠相比,miR-inhibitor组小鼠的骨小梁组织体积(t=3.32,P=0.029)、骨小梁区域体积(t=6.24,P=0.003)、骨体积分数(t=7.35,P=0.002)和骨矿物密度(t=13.72,P<0.001)均增加,成熟破骨细胞数量减少。与NC-inhibitor组比较,miR-inhibitor组细胞中c-fos(t=9.04,P=0.001)和NFATc1(t=13.42,P<0.001)的表达减少。与miR-inhibitor+si-NC组相比,miR-inhibitor+si-PIK3R1组中小鼠的骨小梁组织体积(t=3.03,P=0.039)、骨小梁区域体积(t=6.37,P=0.003)、骨体积分数(t=3.36,P=0.028)和骨矿物密度(t=6.92,P=0.002)均减少,成熟破骨细胞数量增加,细胞中c-fos(t=7.75,P=0.002)和NFATc1(t=9.65,P=0.001)的表达增加。结论ER(+)BC患者血浆外泌体源性miR-20a-5p通过抑制PIK3R1表达来促进ER(+)BC骨转移。展开更多
文摘目的:探讨磷脂酰肌醇-3激酶调节亚基1(phosphoinositide-3-kinase regulatory subunit 1,PIK3R1)及其编码蛋白p85α在宫颈鳞癌中的表达及临床价值。方法:选取38例宫颈鳞癌组织和39例良性宫颈组织,采用HE染色观察鳞癌宫颈和良性宫颈的形态,免疫组织化学法检测p85α蛋白的表达;分析宫颈鳞癌组织中p85α的表达与各临床参数的相关性;选取19例宫颈鳞癌组织和19例良性宫颈组织,采用实时荧光定量PCR(qRT-PCR)技术检测PIK3R1和人第10号染色体缺失的磷酸酶及张力蛋白同源的基因(gene of phosphatase and tension homology deleted on chromsome ten,PTEN)的表达,并分析二者的相关性。结果:与良性宫颈组织相比,p85α蛋白在宫颈鳞癌组织中表达明显降低(P<0.01);病理分期越高,p85α的表达越低(P<0.01),有远处转移的肿瘤组织中p85α的表达较无远处转移的肿瘤组织表达低(P<0.05)。与良性宫颈组织相比,PIK3R1和PTEN在宫颈鳞癌组织中的表达均明显下调(P<0.01)。相关分析表明,PIK3R1和PTEN在宫颈鳞癌中表达呈中度相关(r=0.567,P=0.011)。结论:PIK3R1及编码蛋白p85α在宫颈鳞癌中表达均下调,二者在宫颈鳞癌中可能作为抑癌基因发挥关键作用。
基金financially supported by the National Natural Science Foundation of China(31872330,31802041)Beijing Dairy Industry Innovation Team(BAIC06–2018/2019)+3 种基金Beijing Science and Technology Program(D171100002417001)National Science and Technology Programs of China(2013AA102504)earmarked fund for Modern Agro-industry Technology Research System(CARS-36)the Program for Changjiang Scholar and Innovation Research Team in University(IRT_15R62).
文摘Background:Previously,phosphoinositide-3-kinase regulatory subunit 1(PIK3R1)and dual specificity phosphatase 1(DUSP1)were identified as promising candidate genes for milk production traits due to their being differentially expressed between the dry period and the peak of lactation in livers of dairy cows.Hence,in this study,the single nucleotide polymorphisms(SNPs)of PIK3R1 and DUSP1 genes were identified and their genetic associations with milk yield,fat yield,fat percentage,protein yield,and protein percentage,were investigated using 1067 Chinese Holstein cows from 40 sire families.Results:By re-sequencing the entire coding region and 2000 bp of the 5′and 3′flanking regions of the two genes,one SNP in the 5′untranslated region(UTR),three in the 3′UTR,and two in the 3′flanking region of PIK3R1 were identified,and one in the 5′flanking region,one in the 3′UTR,and two in the 3′flanking region of DUSP1 were found.Subsequent single-locus association analyses showed that five SNPs in PIK3 R1,rs42590258,rs210389799,rs208819656,rs41255622,rs133655926,and rs211408208,and four SNPs in DUSP1,rs207593520,rs208460068,rs209154772,and rs210000760,were significantly associated with milk,fat and protein yields in the first or second lactation(P values≤0.0001 and 0.0461).In addition,by the Haploview 4.2 software,the six and four SNPs in PIK3R1 and DUSP1 respectively formed one haplotype block,and the haplotype-based association analyses showed significant associations between their haplotype combinations and the milk traits in both two lactations(P values≤0.0001 and 0.0364).One SNP,rs207593520(T/G),was predicted to alter the transcription factor binding sites(TFBSs)in the 5′flanking region of DUSP1.Further,the dual-luciferase assay showed that the transcription activity of allele T in rs207593520 was significantly higher than that of allele G,suggesting the activation of transcriptional activity of DUSP1 gene by allele T of rs207593520.Thus,the rs207593520 SNP was highlighted as a potential causal mutation that should be further verified.Conclusions:We demonstrated novel and significant genetic effects of the PIK3R1 and DUSP1 genes on milk production traits in dairy cows,and our findings provide information for use in dairy cattle breeding.
文摘目的探讨血浆外泌体源性miR-20a-5p通过靶向PIK3R1对雌激素受体阳性的乳腺癌(estrogen receptor-positive breast cancer,ER(+)BC)骨转移的影响。方法借助生物信息学网站检索与ER(+)BC骨转移相关的数据集,选择miR-20a-5p纳入研究。收集90例ER(+)BC患者与相对应的健康人的血浆,检测血浆中PIK3R1的表达,从血浆中提取外泌体,检测外泌体中miR-20a-5p的表达。双荧光素酶报告实验验证miR-20a-5p与PIK3R1的靶向调控关系。分别将转染NC-inhibitor、miR-inhibitor的外泌体或转染si-NC、si-PIK3R1的ER(+)BC细胞注射到小鼠左心室,Micro-CT扫描骨组织并进行骨组织TRAP染色。将转染NC-inhibitor、miR-inhibitor的外泌体与转染si-NC、si-PIK3R1的ER(+)BC细胞共培养后,使用Western blot检测破骨细胞分子c-fos、NFATc1的表达。结果qRT-PCR检测发现较正常血浆外泌体(1±0.26)或细胞(1±0.13),miR-20a-5p在ER(+)BC血浆外泌体(1.49±0.27)(t=12.40,P<0.001)及BC细胞系MCF-7(1.64±0.13)(t=6.03,P=0.004)、BT474(1.49±0.11)(t=4.98,P=0.008)、T47D(1.98±0.15)(t=8.55,P=0.001)中均表达升高。而与正常血浆外泌体(1±0.25)或细胞(1±0.10)比较,PIK3R1在ER(+)BC血浆外泌体(0.69±0.24)(t=8.48,P<0.001)及ER(+)BC细胞系MCF-7(0.73±0.05)(t=4.18,P=0.014)、BT474(0.61±0.05)(t=6.04,P=0.004)、T47D(0.34±0.04)(t=10.61,P<0.001)中则表达降低。PIK3R1在血浆(t=8.48,P<0.001)及ER(+)BC细胞系MCF-7(t=4.18,P=0.014)、BT474(t=6.04,P=0.004)、T47D(t=10.61,P<0.001)中则表达降低。PIK3R1被证实为miR-20a-5p的靶基因。与NC-inhibitor组小鼠相比,miR-inhibitor组小鼠的骨小梁组织体积(t=3.32,P=0.029)、骨小梁区域体积(t=6.24,P=0.003)、骨体积分数(t=7.35,P=0.002)和骨矿物密度(t=13.72,P<0.001)均增加,成熟破骨细胞数量减少。与NC-inhibitor组比较,miR-inhibitor组细胞中c-fos(t=9.04,P=0.001)和NFATc1(t=13.42,P<0.001)的表达减少。与miR-inhibitor+si-NC组相比,miR-inhibitor+si-PIK3R1组中小鼠的骨小梁组织体积(t=3.03,P=0.039)、骨小梁区域体积(t=6.37,P=0.003)、骨体积分数(t=3.36,P=0.028)和骨矿物密度(t=6.92,P=0.002)均减少,成熟破骨细胞数量增加,细胞中c-fos(t=7.75,P=0.002)和NFATc1(t=9.65,P=0.001)的表达增加。结论ER(+)BC患者血浆外泌体源性miR-20a-5p通过抑制PIK3R1表达来促进ER(+)BC骨转移。
基金supported by the Projects of Traditional Chinese Medicine Science and Technology(No.2021ZA076,2020ZB105)the Project of Medical and Health Science of Zhejiang Province,China(No.2021KY226)。
