为提供制备抗病转基因山羊核供体细胞,研究Toll样受体9(Toll like receptor 9,TLR9)对山羊抗病性能的影响,应用反转录聚合酶链式反应(RT-PCR)技术,从山羊外周血单核细胞中克隆了TLR9基因,并对其遗传进化关系及分子结构特征进行了分析,...为提供制备抗病转基因山羊核供体细胞,研究Toll样受体9(Toll like receptor 9,TLR9)对山羊抗病性能的影响,应用反转录聚合酶链式反应(RT-PCR)技术,从山羊外周血单核细胞中克隆了TLR9基因,并对其遗传进化关系及分子结构特征进行了分析,发现克隆的基因具有Toll样受体家族的典型结构特征,其遗传进化关系与物种亲缘关系密切相关。将克隆的山羊TLR9基因亚克隆至pIRES2-EGFP真核表达载体,转染山羊胎儿成纤维细胞,经G418筛选以及阳性细胞基因组PCR鉴定和荧光观察证实,外源TLR9基因确已稳定整合入转染阳性细胞克隆基因组中并获得表达。成功建立了稳定表达外源TLR9的山羊胎儿成纤维细胞系,为培育抗病转基因山羊奠定了基础。展开更多
Prostate cancer(PCa)is one of the most frequent cancers in men,and its biomolecular targets have been extensively studied.This study aimed to analyze the expression of toll-like receptor 9(TLR9)and vascular endothelia...Prostate cancer(PCa)is one of the most frequent cancers in men,and its biomolecular targets have been extensively studied.This study aimed to analyze the expression of toll-like receptor 9(TLR9)and vascular endothelial growth factor C(VEGF-C)and the clinical value of the coexpression of TLR9 and VEGF-C in PCa.We retrospectively evaluated 55 patients with clinically localized,intermediate-risk,or high-risk PCa who underwent laparoscopic radical prostatectomy(LRP)and extended pelvic lymph node dissection(ePLND)without neoadjuvant hormonal therapy at a single institution from June 2013 to December 2016.In all 55 patients,the median number of lymph nodes(LNs)resected was 23(range:18-31),and a total of 1269 LNs were removed,of which 78 LNs were positive.Seventeen patients had positive LNs,with a positive rate of 30.9%.In addition,the immunohistochemical results in the above patients revealed that high TLR9 expression was correlated with higher Gleason score(GS)(P=0.049),increased LN metastasis(P=0.004),and more perineural invasion(PNI)(P=0.033).Moreover,VEGF-C expression was associated with GS(P=0.040),pathological stage(pT stage)(P=0.022),LN metastasis(P=0.003),and PNI(P=0.001).Furthermore,a significant positive correlation between TLR9 and VEGF-C was found(P<0.001),and the TLR9/VEGF-C phenotype was associated with LN metastasis(P=0.047).Collectively,we propose that TLR9 stimulation may promote LN metastasis in PCa cells through the upregulation of VEGF-C expression,thereby affecting the prognosis of PCa patients.Therefore,these markers may serve as valuable targets for the treatment of PCa.展开更多
文摘为提供制备抗病转基因山羊核供体细胞,研究Toll样受体9(Toll like receptor 9,TLR9)对山羊抗病性能的影响,应用反转录聚合酶链式反应(RT-PCR)技术,从山羊外周血单核细胞中克隆了TLR9基因,并对其遗传进化关系及分子结构特征进行了分析,发现克隆的基因具有Toll样受体家族的典型结构特征,其遗传进化关系与物种亲缘关系密切相关。将克隆的山羊TLR9基因亚克隆至pIRES2-EGFP真核表达载体,转染山羊胎儿成纤维细胞,经G418筛选以及阳性细胞基因组PCR鉴定和荧光观察证实,外源TLR9基因确已稳定整合入转染阳性细胞克隆基因组中并获得表达。成功建立了稳定表达外源TLR9的山羊胎儿成纤维细胞系,为培育抗病转基因山羊奠定了基础。
基金supported by grants from the Natural Science Foundation of Guangdong Province(No.2018A030313261 and No.2021Al515010129)Guangdong Medical Science and Technology Research Foundation(No.A2018079)the National Natural Science Foundation of China(No.81772752).
文摘Prostate cancer(PCa)is one of the most frequent cancers in men,and its biomolecular targets have been extensively studied.This study aimed to analyze the expression of toll-like receptor 9(TLR9)and vascular endothelial growth factor C(VEGF-C)and the clinical value of the coexpression of TLR9 and VEGF-C in PCa.We retrospectively evaluated 55 patients with clinically localized,intermediate-risk,or high-risk PCa who underwent laparoscopic radical prostatectomy(LRP)and extended pelvic lymph node dissection(ePLND)without neoadjuvant hormonal therapy at a single institution from June 2013 to December 2016.In all 55 patients,the median number of lymph nodes(LNs)resected was 23(range:18-31),and a total of 1269 LNs were removed,of which 78 LNs were positive.Seventeen patients had positive LNs,with a positive rate of 30.9%.In addition,the immunohistochemical results in the above patients revealed that high TLR9 expression was correlated with higher Gleason score(GS)(P=0.049),increased LN metastasis(P=0.004),and more perineural invasion(PNI)(P=0.033).Moreover,VEGF-C expression was associated with GS(P=0.040),pathological stage(pT stage)(P=0.022),LN metastasis(P=0.003),and PNI(P=0.001).Furthermore,a significant positive correlation between TLR9 and VEGF-C was found(P<0.001),and the TLR9/VEGF-C phenotype was associated with LN metastasis(P=0.047).Collectively,we propose that TLR9 stimulation may promote LN metastasis in PCa cells through the upregulation of VEGF-C expression,thereby affecting the prognosis of PCa patients.Therefore,these markers may serve as valuable targets for the treatment of PCa.
