miRNA can regulate development and milk yield of the mammary gland through epigenetic mechanism, miRNA can directly and indirectly modulate the activity of the epigenetic machinery, target genes through post-inhibitio...miRNA can regulate development and milk yield of the mammary gland through epigenetic mechanism, miRNA can directly and indirectly modulate the activity of the epigenetic machinery, target genes through post-inhibition of translation initiation, mediate miRNA decay, target genes and inhibit the positive regulation, regulate tone modification, and regulate DNA methylation of target genes. Here we reviewed the role of miRNAs in mammary gland development and lactation. Researching miRNA in mammary gland development and lactation process, and understanding the response of the epigenetic mechanisms to external stimuli will be an important necessity to devise new technologies for maximizing their activity and milk production in the dairy cow.展开更多
Mammary epithelial cells with lactational function can be a valuable cellular model for research of the development and regulation of the mammary gland.This paper describes some aspects of function of an epithelial ce...Mammary epithelial cells with lactational function can be a valuable cellular model for research of the development and regulation of the mammary gland.This paper describes some aspects of function of an epithelial cell line from the mammary gland of the dairy goat.SDS-PAGE,triglyceride and lactose content of cultured cells were used to assess synthetic function of cells and the effects of exposure to insulin and prolactin.Results show that goat mammary epithelial cells can synthesize fat,proteins and lactose when they were cultured in DMEM-F12 medium with added EGF,IGF-1,ITS and FBS.There were no obvious changes after 48h treatment with additional insulin.Prolactin added to the basal medium significantly increased synthesis of proteins and lactose.A mammary gland epithelial cell line from goats which has lactational function has been established.This outcome provides a valuable and convenient model system.展开更多
To apply gene therapy to breast carcinoma wepropose a model for in vitro gene transfer of the herpessimplex thymidine kinase (HSV-tk) gene using Stkvector-producing cells. In this study the HSV-tk gene wastransferred ...To apply gene therapy to breast carcinoma wepropose a model for in vitro gene transfer of the herpessimplex thymidine kinase (HSV-tk) gene using Stkvector-producing cells. In this study the HSV-tk gene wastransferred to mouse mammary carcinoma cell line 4Tlwith the method of infection. The HSV-tk gene in thegenome of transduced cells (4Tl/tk) was demonstratedby the method of DNA-PCR. Using a modified展开更多
The aim of this study was to investigate the effect of duodenal infusion of soybean small peptides(SSP) on mammary uptake of amino acids(AA).Eight Chinese Saanen goats with duodenal fistulae and catheters were used in...The aim of this study was to investigate the effect of duodenal infusion of soybean small peptides(SSP) on mammary uptake of amino acids(AA).Eight Chinese Saanen goats with duodenal fistulae and catheters were used in a crossover design trial.Goats were infused with 0,60,120,180 g/d SSP.The experimental period lasted for 14 d.The results showed:1) milk protein yield and content were increased,with the increment in milk protein yield being significant(P【0.05).Milk fat yield and content were decreased with the increased amount of SSP infused(P【0.05).2) Mammary plasma flow was not changed(P】0.05) by SSP infusion though the average was slightly higher.Mammary plasma flow/milk yield was decreased by SSP infusion and there was significant difference between the 120 g/d treatment and the control(P【0.05).3) Compared with the control treatment,uptakes of most free amino acids were increased in the 60 and 120 g/d treatments,but decreased by the 180 g/d treatment.The uptakes of all peptide-bound essential amino acids(PB-EAA) were increased except for PB-Ile.Uptake of PB-Val,PB-Leu, PB-Phe,PB-Thr,PB-Met and PB-Lys was highest in the 120 g/d treatment.Among the peptide-bound nonessential amino acids(PB-NEAA),uptake of PB-Ser,PB-Tyr,and PB-Pro was increased(P【0.05) while that of PB-Gly was decreased(P】0.05).4) With the exception of Lys,secretion of all essential AA(EAA) was increased by SSP infusion from 0-120 g/d(P【0.05),while in the 180 g/d treatment the increase was not significant and was lower than that in the 120 g/d treatment.5) The expression of APN was increased by SSP;in the 60,120,180 g/d treatments expression was 13.55-,18.69-,and 10.01-fold over that of the controls.Conclusion:SSP might be used as substrates for milk protein synthesis by mammary gland,and can increase synthesis and secretion of milk protein.There was reason to believe that uptake and use of AA was either saturated or inhibited at high levels of infusion of SSP.Increased APN expression with infusion of SSP related to mammary uptake of PBAA by mammary gland,which suggested that APN was one of the peptidases regulating use of AA from small peptides in mammary tissue.展开更多
To investigate the expression of antibacterial peptide CecropinB cDNA in dairy goat mammary gland epithelial cells,the CecropinB gene was cloned and was inserted into a eukaryotic vector pECFP-C1 to construct the reco...To investigate the expression of antibacterial peptide CecropinB cDNA in dairy goat mammary gland epithelial cells,the CecropinB gene was cloned and was inserted into a eukaryotic vector pECFP-C1 to construct the recombinant plasmid pECFP-B by genetic engineering technique.Recombinant plasmid pECFP-B was transfected into dairy goat mammary gland epithelial to detect the bactericidal activity of CecropinB.The expression of CecropinB was also detected.The result of RT-PCR demonstrated CecropinB gene was expressed in transfected cells.CecropinB recombinant plasmid DNA was injected into udders and CecropinB was expressed in mammary gland,exhibiting bactericidal activity to Staphylococcus aureus in vivo experiments.展开更多
The paper aimed at constructing the eukaryotic expression vector of CecropinB and transfecting it into the goat mammary epithelial cell line. The recombinant plasmid inserted with CecropinB was transfected into the go...The paper aimed at constructing the eukaryotic expression vector of CecropinB and transfecting it into the goat mammary epithelial cell line. The recombinant plasmid inserted with CecropinB was transfected into the goat mammary epithelial cell by liposome Tfx^M-20. By screening with G418, the stable transfected goat mammary epithelial cell line was established and the transcription and expression of CecropinB were identified by RT-PCR and agarose diffusion experiment, respectively. Results showed that eukaryotic expression vector pECFP-B was constructed successfully. Stable transfected goat mammary epithelial cell line was established and the CecropinB protein was expressed successfully. It provides the solid foundation for further experimental studies on the function of CecropinB.展开更多
Leptin, a protein hormone produced and secreted predominantly by white adipose tissue, has a critical role in the regulation and coordination of energy metabolism. Leptin is produced in the mammary gland by the fat ti...Leptin, a protein hormone produced and secreted predominantly by white adipose tissue, has a critical role in the regulation and coordination of energy metabolism. Leptin is produced in the mammary gland by the fat tissue or by the mammary epithelium. In vitro study has shown that leptin triggers apoptosis in mammary epithelial cells. Mammary gland involution is characterized by extensive apoptosis of the epithelial cells. At the onset of involution, STAT3 is specifically activated.Various studies show that leptin act as a paracrine and autocrin factor to influence mammary epithelial cell proliferation and differentiation. This paper reviewed the function of leptin to the involution of mammary gland.展开更多
The lactating mammary gland is a prodigious protein-producing factory, but the milk protein synthesis mechanisms are not well understood. The major objective of this paper was to elucidate which genes and pathways wer...The lactating mammary gland is a prodigious protein-producing factory, but the milk protein synthesis mechanisms are not well understood. The major objective of this paper was to elucidate which genes and pathways were involved in the regulation of milk protein synthesis in the dairy goat mammary gland. Total 36 primiparous Guanzhong dairy goats were allotted in 12 groups according to their mammary development stages： days 90 and 150 of virgin, days 30, 90, and 150 of pregnancy, days 1, 10, 35, and 60 of lactation and days 3, 7, and 21 of involution （three animals per group）. Mammary tissue RNA was isolated for quantitative real- time RT-PCR of four casein genes alpha-s 1 casein （CSN 1S 1 ）, alpha-s2 casein （CSN 1 S2）, beta-casein （CSN2） and casein kappa （CSN3）, four whey protein genes lactoglobulin （LGB）, laetalbumin （LALBA）, laetofarrin （LTF）, and Whey acidic protein （WAP） and the genes which were potentially to regulate dairy goat milk protein synthesis at the level of transcription or translation [prolactin receptor （PRLR）, AKT1, signal transducers and activators of transcription 5 （STAT5）, E74-Like Factor 5 （ELF5）, eukaryotic translation initiation factor 4E binding protein 1 （EIF4E-BP1）, S6kinase （S6K） and caveolin 1]. The results showed that all genes were up-regulated in lactation period. The expressions of PRLR, AKT1, STAT5, ELF5, and S6K were similar to mRNA expressions of milk proteins. Our results indicated that milk protein synthesis in dairy goat mammary gland was possibly regulated by these genes.展开更多
let-7g, a member of the let-7 family, regulates gene expression at the post-transcriptional level. The study explored a series of biological effects of mouse mammary epithelial cells that let-7g was produced. The diff...let-7g, a member of the let-7 family, regulates gene expression at the post-transcriptional level. The study explored a series of biological effects of mouse mammary epithelial cells that let-7g was produced. The differential expression of let-7g was detected by qRT-PCR in different developmental stages of the mouse mammary gland, let-7g expression and impact of let-7g on mouse mammary epithelial cells were analyzed by CASY-technology, qRT-PCR, Western blotting and HPLC inhibited let-7g expression of mouse mammary epithelial ceils through gene silencing. The results showed that qRT-PCR identified let-7g as being down-regulated in mouse mammary epithelial cells after it was inhibited. Mouse mammary epithelial cells with low expression of let-7g displayed higher expression of TGFβR I protein than those with high expression of let-7g, suggesting that low let-7g expression contributed to TGFβR I over-expression. Finally, the expression of let-7g was down-regulated, which significantly enhanced the proliferation of mouse mammary epithelial cells, and increased expression of β-Casein. The data indicated that let-7g could negatively regulate the expression of target Tgfbrl by complementary combination in mouse mammary epithelial cells, and then regulate the cell proliferation and expression of β-Casein by suppressing the TGFβR I expression.展开更多
A galactopoietic compound, identified as dibutyl phthalate（DBP）, was isolated from Vaccaria segetalis. The activity of DBP on lactation ability of dairy cow mammary gland epithelial cells（DCMECs） cultured in vitro...A galactopoietic compound, identified as dibutyl phthalate（DBP）, was isolated from Vaccaria segetalis. The activity of DBP on lactation ability of dairy cow mammary gland epithelial cells（DCMECs） cultured in vitro and dairy cow was evaluated. Results showed that DBP could promote cell viability, proliferation ability, lactose and β-casein secretion of DCMECs, which could also raise the milk yields of dairy cows significantly.展开更多
In order to investigate the possible relationshipbetween the transfection of IL-2 gene and the changes intumor cells biological characteristics, we used retroviralvector to transduce human mammary carcinoma MCF-7cell ...In order to investigate the possible relationshipbetween the transfection of IL-2 gene and the changes intumor cells biological characteristics, we used retroviralvector to transduce human mammary carcinoma MCF-7cell line with the IL-2 cDNA and explored the changes incell membrane components and function. The genemodified cells releasing 70 - 169 U/106 cells of IL-2 in 24hrs. showed apparent morphological conversion includingthe appearance change from epitheloid form展开更多
To analyze miR-139 target sites in 3＇ UTR of GHR gene in dairy cow mammary gland, a GHR 3＇ UTR- luciferase reporter vector was constructed and the effect of miRNA on its activity was evaluated in dairy cow mammary g...To analyze miR-139 target sites in 3＇ UTR of GHR gene in dairy cow mammary gland, a GHR 3＇ UTR- luciferase reporter vector was constructed and the effect of miRNA on its activity was evaluated in dairy cow mammary gland epithelial cells （DCMECs）. The miR-139 targeting GHR 3＇ UTR was predicted by Target Scan 5.1 software, 3＇ UTR fragment of GHR was amplified by PCR from RNA of DCMECs. PCR products were cloned into Spe Ⅰ/Hind Ⅱ modified pMIR-Report vector. The luciferase reporter vector and miRNA eukaryotic expression vector were transferred into DCMECs using lipofectamine 2000 transfection reagent. The dualluciferase reporter assay system was used to quantitiate the reporter activity. The results showed that a 107 bp 3＇ UTR fragment of GHR gene was successfully cloned into the pMIR-Report vector, which authenticated by Spe Ⅰ/Hind Ⅲ digestion and DNA sequencing. The luciferase activity of reporter construction treated with miR-139 decreased 20.87% compared with the control group. It was concluded that the GHR3＇ UTR-luciferase reporter vector had been successfully constructed. The luciferase activity of the reporter could be suppressed by miR- 139.展开更多
基金Support by the Natural Science Foundation of China(31072103)
文摘miRNA can regulate development and milk yield of the mammary gland through epigenetic mechanism, miRNA can directly and indirectly modulate the activity of the epigenetic machinery, target genes through post-inhibition of translation initiation, mediate miRNA decay, target genes and inhibit the positive regulation, regulate tone modification, and regulate DNA methylation of target genes. Here we reviewed the role of miRNAs in mammary gland development and lactation. Researching miRNA in mammary gland development and lactation process, and understanding the response of the epigenetic mechanisms to external stimuli will be an important necessity to devise new technologies for maximizing their activity and milk production in the dairy cow.
基金supported by Innovation Team Project of Northeast Agricultural Vniversity(XLT005-1-2)Research Fund for the Doctoral Program of Heilongjiang Educational Committee(HLJBSDJI2004-15)
文摘Mammary epithelial cells with lactational function can be a valuable cellular model for research of the development and regulation of the mammary gland.This paper describes some aspects of function of an epithelial cell line from the mammary gland of the dairy goat.SDS-PAGE,triglyceride and lactose content of cultured cells were used to assess synthetic function of cells and the effects of exposure to insulin and prolactin.Results show that goat mammary epithelial cells can synthesize fat,proteins and lactose when they were cultured in DMEM-F12 medium with added EGF,IGF-1,ITS and FBS.There were no obvious changes after 48h treatment with additional insulin.Prolactin added to the basal medium significantly increased synthesis of proteins and lactose.A mammary gland epithelial cell line from goats which has lactational function has been established.This outcome provides a valuable and convenient model system.
文摘To apply gene therapy to breast carcinoma wepropose a model for in vitro gene transfer of the herpessimplex thymidine kinase (HSV-tk) gene using Stkvector-producing cells. In this study the HSV-tk gene wastransferred to mouse mammary carcinoma cell line 4Tlwith the method of infection. The HSV-tk gene in thegenome of transduced cells (4Tl/tk) was demonstratedby the method of DNA-PCR. Using a modified
文摘The aim of this study was to investigate the effect of duodenal infusion of soybean small peptides(SSP) on mammary uptake of amino acids(AA).Eight Chinese Saanen goats with duodenal fistulae and catheters were used in a crossover design trial.Goats were infused with 0,60,120,180 g/d SSP.The experimental period lasted for 14 d.The results showed:1) milk protein yield and content were increased,with the increment in milk protein yield being significant(P【0.05).Milk fat yield and content were decreased with the increased amount of SSP infused(P【0.05).2) Mammary plasma flow was not changed(P】0.05) by SSP infusion though the average was slightly higher.Mammary plasma flow/milk yield was decreased by SSP infusion and there was significant difference between the 120 g/d treatment and the control(P【0.05).3) Compared with the control treatment,uptakes of most free amino acids were increased in the 60 and 120 g/d treatments,but decreased by the 180 g/d treatment.The uptakes of all peptide-bound essential amino acids(PB-EAA) were increased except for PB-Ile.Uptake of PB-Val,PB-Leu, PB-Phe,PB-Thr,PB-Met and PB-Lys was highest in the 120 g/d treatment.Among the peptide-bound nonessential amino acids(PB-NEAA),uptake of PB-Ser,PB-Tyr,and PB-Pro was increased(P【0.05) while that of PB-Gly was decreased(P】0.05).4) With the exception of Lys,secretion of all essential AA(EAA) was increased by SSP infusion from 0-120 g/d(P【0.05),while in the 180 g/d treatment the increase was not significant and was lower than that in the 120 g/d treatment.5) The expression of APN was increased by SSP;in the 60,120,180 g/d treatments expression was 13.55-,18.69-,and 10.01-fold over that of the controls.Conclusion:SSP might be used as substrates for milk protein synthesis by mammary gland,and can increase synthesis and secretion of milk protein.There was reason to believe that uptake and use of AA was either saturated or inhibited at high levels of infusion of SSP.Increased APN expression with infusion of SSP related to mammary uptake of PBAA by mammary gland,which suggested that APN was one of the peptidases regulating use of AA from small peptides in mammary tissue.
基金Supported by Key Teachers Foundation of Education Office of Heilongjiang Province,2005(1055G005)
文摘To investigate the expression of antibacterial peptide CecropinB cDNA in dairy goat mammary gland epithelial cells,the CecropinB gene was cloned and was inserted into a eukaryotic vector pECFP-C1 to construct the recombinant plasmid pECFP-B by genetic engineering technique.Recombinant plasmid pECFP-B was transfected into dairy goat mammary gland epithelial to detect the bactericidal activity of CecropinB.The expression of CecropinB was also detected.The result of RT-PCR demonstrated CecropinB gene was expressed in transfected cells.CecropinB recombinant plasmid DNA was injected into udders and CecropinB was expressed in mammary gland,exhibiting bactericidal activity to Staphylococcus aureus in vivo experiments.
基金Supported by Key Teachers Foundation of Education Office of Heilongjiang Province, 2005 (1055G005)
文摘The paper aimed at constructing the eukaryotic expression vector of CecropinB and transfecting it into the goat mammary epithelial cell line. The recombinant plasmid inserted with CecropinB was transfected into the goat mammary epithelial cell by liposome Tfx^M-20. By screening with G418, the stable transfected goat mammary epithelial cell line was established and the transcription and expression of CecropinB were identified by RT-PCR and agarose diffusion experiment, respectively. Results showed that eukaryotic expression vector pECFP-B was constructed successfully. Stable transfected goat mammary epithelial cell line was established and the CecropinB protein was expressed successfully. It provides the solid foundation for further experimental studies on the function of CecropinB.
基金Supported by National Natural Sciences Foundation(30671538)
文摘Leptin, a protein hormone produced and secreted predominantly by white adipose tissue, has a critical role in the regulation and coordination of energy metabolism. Leptin is produced in the mammary gland by the fat tissue or by the mammary epithelium. In vitro study has shown that leptin triggers apoptosis in mammary epithelial cells. Mammary gland involution is characterized by extensive apoptosis of the epithelial cells. At the onset of involution, STAT3 is specifically activated.Various studies show that leptin act as a paracrine and autocrin factor to influence mammary epithelial cell proliferation and differentiation. This paper reviewed the function of leptin to the involution of mammary gland.
基金Supported by the National Basic Research Program of China(973 Program,2011CB100804)the National Natural Science Foundation of China(31101784)Funds for Young Researchers from Northeast Agricultural University(14QC43)
文摘The lactating mammary gland is a prodigious protein-producing factory, but the milk protein synthesis mechanisms are not well understood. The major objective of this paper was to elucidate which genes and pathways were involved in the regulation of milk protein synthesis in the dairy goat mammary gland. Total 36 primiparous Guanzhong dairy goats were allotted in 12 groups according to their mammary development stages： days 90 and 150 of virgin, days 30, 90, and 150 of pregnancy, days 1, 10, 35, and 60 of lactation and days 3, 7, and 21 of involution （three animals per group）. Mammary tissue RNA was isolated for quantitative real- time RT-PCR of four casein genes alpha-s 1 casein （CSN 1S 1 ）, alpha-s2 casein （CSN 1 S2）, beta-casein （CSN2） and casein kappa （CSN3）, four whey protein genes lactoglobulin （LGB）, laetalbumin （LALBA）, laetofarrin （LTF）, and Whey acidic protein （WAP） and the genes which were potentially to regulate dairy goat milk protein synthesis at the level of transcription or translation [prolactin receptor （PRLR）, AKT1, signal transducers and activators of transcription 5 （STAT5）, E74-Like Factor 5 （ELF5）, eukaryotic translation initiation factor 4E binding protein 1 （EIF4E-BP1）, S6kinase （S6K） and caveolin 1]. The results showed that all genes were up-regulated in lactation period. The expressions of PRLR, AKT1, STAT5, ELF5, and S6K were similar to mRNA expressions of milk proteins. Our results indicated that milk protein synthesis in dairy goat mammary gland was possibly regulated by these genes.
基金Supported by the National Natural Science Foundation (31072103)
文摘let-7g, a member of the let-7 family, regulates gene expression at the post-transcriptional level. The study explored a series of biological effects of mouse mammary epithelial cells that let-7g was produced. The differential expression of let-7g was detected by qRT-PCR in different developmental stages of the mouse mammary gland, let-7g expression and impact of let-7g on mouse mammary epithelial cells were analyzed by CASY-technology, qRT-PCR, Western blotting and HPLC inhibited let-7g expression of mouse mammary epithelial ceils through gene silencing. The results showed that qRT-PCR identified let-7g as being down-regulated in mouse mammary epithelial cells after it was inhibited. Mouse mammary epithelial cells with low expression of let-7g displayed higher expression of TGFβR I protein than those with high expression of let-7g, suggesting that low let-7g expression contributed to TGFβR I over-expression. Finally, the expression of let-7g was down-regulated, which significantly enhanced the proliferation of mouse mammary epithelial cells, and increased expression of β-Casein. The data indicated that let-7g could negatively regulate the expression of target Tgfbrl by complementary combination in mouse mammary epithelial cells, and then regulate the cell proliferation and expression of β-Casein by suppressing the TGFβR I expression.
基金Supported by "863" Project of Ministry of Science and Technology of China(2013AA102504-03)Talents Foundation of Northeast Agricultural University(2010RCB47,2010RCB55)
文摘A galactopoietic compound, identified as dibutyl phthalate（DBP）, was isolated from Vaccaria segetalis. The activity of DBP on lactation ability of dairy cow mammary gland epithelial cells（DCMECs） cultured in vitro and dairy cow was evaluated. Results showed that DBP could promote cell viability, proliferation ability, lactose and β-casein secretion of DCMECs, which could also raise the milk yields of dairy cows significantly.
文摘In order to investigate the possible relationshipbetween the transfection of IL-2 gene and the changes intumor cells biological characteristics, we used retroviralvector to transduce human mammary carcinoma MCF-7cell line with the IL-2 cDNA and explored the changes incell membrane components and function. The genemodified cells releasing 70 - 169 U/106 cells of IL-2 in 24hrs. showed apparent morphological conversion includingthe appearance change from epitheloid form
基金Supported by the National Key Basic Research and Development (973) Plan (2011CB100804)Northeast Agricultural University Innovation Team Project (CXT005-1-1/CXT005-1-2)
文摘To analyze miR-139 target sites in 3＇ UTR of GHR gene in dairy cow mammary gland, a GHR 3＇ UTR- luciferase reporter vector was constructed and the effect of miRNA on its activity was evaluated in dairy cow mammary gland epithelial cells （DCMECs）. The miR-139 targeting GHR 3＇ UTR was predicted by Target Scan 5.1 software, 3＇ UTR fragment of GHR was amplified by PCR from RNA of DCMECs. PCR products were cloned into Spe Ⅰ/Hind Ⅱ modified pMIR-Report vector. The luciferase reporter vector and miRNA eukaryotic expression vector were transferred into DCMECs using lipofectamine 2000 transfection reagent. The dualluciferase reporter assay system was used to quantitiate the reporter activity. The results showed that a 107 bp 3＇ UTR fragment of GHR gene was successfully cloned into the pMIR-Report vector, which authenticated by Spe Ⅰ/Hind Ⅲ digestion and DNA sequencing. The luciferase activity of reporter construction treated with miR-139 decreased 20.87% compared with the control group. It was concluded that the GHR3＇ UTR-luciferase reporter vector had been successfully constructed. The luciferase activity of the reporter could be suppressed by miR- 139.
