Spermatogonial stem cells （SSCs） divide continuously to support spermatogenesis throughout postnatal life and transmit genetic information to the next generation. Here, we report the successful establishment of the ...Spermatogonial stem cells （SSCs） divide continuously to support spermatogenesis throughout postnatal life and transmit genetic information to the next generation. Here, we report the successful establishment of the method for the isolation and identification of human SSCs from testicular tissue, and to determine the culture conditions required to expand SSCs on human embryonic stem cell-derived fibroblast-like cells （hdFs）. Large-scale cultures of SSCs were maintained on hdF feeder layers and expanded in the presence of a combination of cytokines and glial cell line-derived neurotrophic factor for at least 2 months. Cell surface marker analysis showed that SSCs retained high levels of alkaline phosphatase activity and stained strongly for anti-stage-specific embryonic antigen （SSEA）-1, OCT4 and CD49f. They also expressed the genes OCT4, SOX3 and STRA8 as detected by reverse transcription polymerase chain reaction （RT-PCR） analysis. These data clearly illustrate a novel approach for the growth of human SSCs using hdFs as feeder cells, potentially eliminating xenogeneic contaminants. This system provides a new opportunity for the study of the regulatory mechanism of the ‘niche＇ that governs SSC self-renewal, and will be a valuable source of SSCs for potential clinical applications.展开更多
The androgen receptor （AR） plays a critical role in prostate cancer development and progression. This study aimed to use a computerized docking approach to examine the interactions between the human AR and phytooest...The androgen receptor （AR） plays a critical role in prostate cancer development and progression. This study aimed to use a computerized docking approach to examine the interactions between the human AR and phytooestrogens （genistein, daidzein, and flavone） and xeno-oestrogens （bisphenol A, 4-nonylphenol, dichlorodiphenyl trichloroethane [DDT], diethylstilbestrol [DES]）. The predicted three-dimensional structure of AR and androgens was established using X-ray diffraction. The binding of four xeno-oestrogens and three phyto-oestrogens to AR was analysed. The steroids estradiol and dihydrotestosterone （DHT） were used as positive controls and thyroxine as negative control. All the ligands shared the same binding site except for thyroxine. The endogenous hormones DHT and 17β-oestradiol showed the strongest binding with the lowest affinity energy （〈 -10 kcal mol-1）. All three phyto- oestrogens and two xeno-oestrogens （bisphenol A and DES） showed strong binding to AR. The affinities offlavone, genistein, and daidzein were between -8.8 and -8.5 kcal mol 1, while that of bisphenol A was -8.1 kcal mol-l and DES -8.3 kcal mol-1. Another two xeno-oestrogens, 4-nonylphenol and DDT, although they fit within the binding domain of AR, showed weak affinity （-6.4 and -6.7 kcal mol 1, respectively）. The phyto-oestrogens genistein, daidzein and flavone, and the xeno-oestrogens bisphenol A and DES can be regarded as androgenic effectors. The xenooestrogens DDT and 4-nonylphenol bind only weakly to AR.展开更多
Functional nucleic acids(FNAs)refer to a type of oligonucleotides with functions over the traditional genetic roles of nucleic acids,which have been widely applied in screening,sensing and imaging fields.However,the p...Functional nucleic acids(FNAs)refer to a type of oligonucleotides with functions over the traditional genetic roles of nucleic acids,which have been widely applied in screening,sensing and imaging fields.However,the potential application of FNAs in biomedical field is still restricted by the unsatisfactory stability,biocompatibility,biodistribution and immunity of natural nucleic acids(DNA/RNA).Xeno nucleic acids(XNAs)are a kind of nucleic acid analogues with chemically modified sugar groups that possess improved biological properties,including improved biological stability,increased binding affinity,reduced immune responses,and enhanced cell penetration or tissue specificity.In the last two decades,scientists have made great progress in the research of functional xeno nucleic acids,which makes it an emerging attractive biomedical application material.In this review,we summarized the design of functional xeno nucleic acids and their applications in the biomedical field.展开更多
We cloned the three androgen response elements（AREs, including AREI, AREII, and AREIII ） with a core transactivation TATA element of the prostate-specific antigen（PSA） promoter into pGL2 basic vector to create an ...We cloned the three androgen response elements（AREs, including AREI, AREII, and AREIII ） with a core transactivation TATA element of the prostate-specific antigen（PSA） promoter into pGL2 basic vector to create an artificial pGL2/AREs-TATA reporter system, which was applied to evaluating the effects of different xeno- oestrogens[bisphenol A（BPA）, 4-nonylphenol（4-NP）, dichlorodiphenyl trichloroethane（DDT） or diethylstilbestrol （DES）] on androgen receptor（AR） abnormal activation to regulate PSA expression and cell proliferation. In all the three AREs, AREIII-TATA displayed as a major element responsive to AR-mediated DHT stimulation of PSA promoter. Therefore, pGL2/AREIII-TATA reporter was adopted to analyze the activation capacity of AR activated by four different xeno-oestrogens. The activation of pGL2/AREIII-TATA reporter by each xeno-oestrogen was analyzed in two different cell lines, one was HEK293T（Human Embryonic Kidney 293T） cell line, and the other was AR stably expressed DU145 cell line, which was produced by infecting AR with pLenti-puro-AR into the prostate cancer DU145 cells and that were scanned with puromycin and tested by AR antibody. In both the two cell lines, BPA or DES significantly induced AR-mediated transcriptional activity of AREIII-TATA reporter, whereas DDT or 4-nonylphenol did not. Moreover, AR-mediated cell proliferation in response to each of four xeno-oestrogens was measured in MTT assays in both HEK293T cell or AR stably expressed DUI45 cell lines. BPA or DES, as an AR inducer, exhibited an enhanced effect in cell proliferation, rather than the effect of DDT or 4-NP, in both cell lines. Finally, we demonstrated that BPA or DES stimulated PSA expression and enhanced the recruitment of AR onto the PSA promoter, resulting in stronger binding to AREIII sites. Taken together, four xeno-oestrogens were identified to have different activities on AR. BPA and DES are demonstrated to be androgenic effectors in the regulation of PSA activation or cell proliferation.展开更多
文摘Spermatogonial stem cells （SSCs） divide continuously to support spermatogenesis throughout postnatal life and transmit genetic information to the next generation. Here, we report the successful establishment of the method for the isolation and identification of human SSCs from testicular tissue, and to determine the culture conditions required to expand SSCs on human embryonic stem cell-derived fibroblast-like cells （hdFs）. Large-scale cultures of SSCs were maintained on hdF feeder layers and expanded in the presence of a combination of cytokines and glial cell line-derived neurotrophic factor for at least 2 months. Cell surface marker analysis showed that SSCs retained high levels of alkaline phosphatase activity and stained strongly for anti-stage-specific embryonic antigen （SSEA）-1, OCT4 and CD49f. They also expressed the genes OCT4, SOX3 and STRA8 as detected by reverse transcription polymerase chain reaction （RT-PCR） analysis. These data clearly illustrate a novel approach for the growth of human SSCs using hdFs as feeder cells, potentially eliminating xenogeneic contaminants. This system provides a new opportunity for the study of the regulatory mechanism of the ‘niche＇ that governs SSC self-renewal, and will be a valuable source of SSCs for potential clinical applications.
基金This study was supported by Ministry of Science and Technology （No. 2010DFA31430）, the National Natural Science Foundation of China （No. 30871301, 30700827）, Ministry of Education of China （No. 108047）, Jilin Provincial Science ＆ Technology Department （No. 20070719, 20080731, 200905116）. We thank Mr Michael Hoyt, who critically read and revised our manuscript.
文摘The androgen receptor （AR） plays a critical role in prostate cancer development and progression. This study aimed to use a computerized docking approach to examine the interactions between the human AR and phytooestrogens （genistein, daidzein, and flavone） and xeno-oestrogens （bisphenol A, 4-nonylphenol, dichlorodiphenyl trichloroethane [DDT], diethylstilbestrol [DES]）. The predicted three-dimensional structure of AR and androgens was established using X-ray diffraction. The binding of four xeno-oestrogens and three phyto-oestrogens to AR was analysed. The steroids estradiol and dihydrotestosterone （DHT） were used as positive controls and thyroxine as negative control. All the ligands shared the same binding site except for thyroxine. The endogenous hormones DHT and 17β-oestradiol showed the strongest binding with the lowest affinity energy （〈 -10 kcal mol-1）. All three phyto- oestrogens and two xeno-oestrogens （bisphenol A and DES） showed strong binding to AR. The affinities offlavone, genistein, and daidzein were between -8.8 and -8.5 kcal mol 1, while that of bisphenol A was -8.1 kcal mol-l and DES -8.3 kcal mol-1. Another two xeno-oestrogens, 4-nonylphenol and DDT, although they fit within the binding domain of AR, showed weak affinity （-6.4 and -6.7 kcal mol 1, respectively）. The phyto-oestrogens genistein, daidzein and flavone, and the xeno-oestrogens bisphenol A and DES can be regarded as androgenic effectors. The xenooestrogens DDT and 4-nonylphenol bind only weakly to AR.
基金supported by the National Natural Science Foundation of China(Nos.22122403,21977027,21890744)the Natural Science Foundation of Hunan Province,China(Nos.2021JJ10012,2019RS1031)。
文摘Functional nucleic acids(FNAs)refer to a type of oligonucleotides with functions over the traditional genetic roles of nucleic acids,which have been widely applied in screening,sensing and imaging fields.However,the potential application of FNAs in biomedical field is still restricted by the unsatisfactory stability,biocompatibility,biodistribution and immunity of natural nucleic acids(DNA/RNA).Xeno nucleic acids(XNAs)are a kind of nucleic acid analogues with chemically modified sugar groups that possess improved biological properties,including improved biological stability,increased binding affinity,reduced immune responses,and enhanced cell penetration or tissue specificity.In the last two decades,scientists have made great progress in the research of functional xeno nucleic acids,which makes it an emerging attractive biomedical application material.In this review,we summarized the design of functional xeno nucleic acids and their applications in the biomedical field.
文摘We cloned the three androgen response elements（AREs, including AREI, AREII, and AREIII ） with a core transactivation TATA element of the prostate-specific antigen（PSA） promoter into pGL2 basic vector to create an artificial pGL2/AREs-TATA reporter system, which was applied to evaluating the effects of different xeno- oestrogens[bisphenol A（BPA）, 4-nonylphenol（4-NP）, dichlorodiphenyl trichloroethane（DDT） or diethylstilbestrol （DES）] on androgen receptor（AR） abnormal activation to regulate PSA expression and cell proliferation. In all the three AREs, AREIII-TATA displayed as a major element responsive to AR-mediated DHT stimulation of PSA promoter. Therefore, pGL2/AREIII-TATA reporter was adopted to analyze the activation capacity of AR activated by four different xeno-oestrogens. The activation of pGL2/AREIII-TATA reporter by each xeno-oestrogen was analyzed in two different cell lines, one was HEK293T（Human Embryonic Kidney 293T） cell line, and the other was AR stably expressed DU145 cell line, which was produced by infecting AR with pLenti-puro-AR into the prostate cancer DU145 cells and that were scanned with puromycin and tested by AR antibody. In both the two cell lines, BPA or DES significantly induced AR-mediated transcriptional activity of AREIII-TATA reporter, whereas DDT or 4-nonylphenol did not. Moreover, AR-mediated cell proliferation in response to each of four xeno-oestrogens was measured in MTT assays in both HEK293T cell or AR stably expressed DUI45 cell lines. BPA or DES, as an AR inducer, exhibited an enhanced effect in cell proliferation, rather than the effect of DDT or 4-NP, in both cell lines. Finally, we demonstrated that BPA or DES stimulated PSA expression and enhanced the recruitment of AR onto the PSA promoter, resulting in stronger binding to AREIII sites. Taken together, four xeno-oestrogens were identified to have different activities on AR. BPA and DES are demonstrated to be androgenic effectors in the regulation of PSA activation or cell proliferation.
