背景:溶质载体家族1成员5(solute carrier family 1 member 5,SLC1A5)在多种疾病中发挥了潜在作用,但确切作用机制尚不清楚。构建稳定的SLC1A5过表达和敲低细胞模型可为深入研究SLC1A5在疾病中的确切作用机制以及发现潜在治疗靶点提供...背景:溶质载体家族1成员5(solute carrier family 1 member 5,SLC1A5)在多种疾病中发挥了潜在作用,但确切作用机制尚不清楚。构建稳定的SLC1A5过表达和敲低细胞模型可为深入研究SLC1A5在疾病中的确切作用机制以及发现潜在治疗靶点提供有力的实验工具。目的:构建小鼠SLC1A5过表达和敲低的慢病毒载体,以建立稳定转染的RAW264.7细胞株,为深入探讨SLC1A5在炎症中的作用提供实验基础。方法:根据SLC1A5基因序列设计合成引物并使用聚合酶链反应扩增该基因片段。将目的基因定向接入经Age I/Nhe I酶切的载体质粒GV492中构建重组慢病毒质粒,对阳性克隆进一步筛选后测序比对结果;pHelper1.0质粒载体、pHelper2.0质粒载体、目的质粒载体与293T细胞共同培养并转染,获得慢病毒原液进行包装和滴度测定;在此基础上,通过体外培养RAW264.7细胞,确定嘌呤霉素工作质量浓度;不同滴度的慢病毒分别与RAW264.7细胞共同培养,根据荧光强度确定转染效率;用嘌呤霉素挑选出稳定转染细胞,实时荧光定量聚合酶链反应和蛋白免疫印迹方法检测稳定转染细胞株的SLC1A5基因和蛋白表达水平。结果与结论:(1)测序序列与目的序列一致提示重组慢病毒载体构建成功;(2)过表达SLC1A5慢病毒的滴度为1×10~9 TU/mL,敲低SLC1A5慢病毒的滴度为3×10~9 TU/mL;(3)确定RAW264.7细胞嘌呤霉素工作质量浓度为3μg/mL;(4)过表达/敲低SLC1A5慢病毒转染RAW264.7细胞的最佳条件皆为HiTransG P转染增强液且感染复数值等于50;(5)过表达SLC1A5稳转细胞株中SLC1A5基因和蛋白的表达量明显上调,而敲低SLC1A5稳转细胞株中SLC1A5基因和蛋白的表达量显著下调。结果表明,成功构建了小鼠SLC1A5过表达和敲低的慢病毒载体并获得稳定转染的RAW264.7细胞株。展开更多
Background：H syndrome （OMIM 612391） is a recently described autosomal recessive genodermatosis characterized by indurated hyperpigmented and hypertrichotic skin,as well as other systemic manifestations.Most of the ...Background：H syndrome （OMIM 612391） is a recently described autosomal recessive genodermatosis characterized by indurated hyperpigmented and hypertrichotic skin,as well as other systemic manifestations.Most of the cases occurred in the Middle East areas or nearby countries such as Spain or India.The syndrome is caused by mutations in solute carrier family 29,member 3 （SLC29A3）,the gene encoding equilibrative nucleoside transporter 3.The aim of this study was to identify pathogenic SLC29A 3 mutations in a Chinese patient clinically diagnosed with H syndrome.Methods：Peripheral blood samples were collected from the patient and his parents.Genomic DNA was isolated by the standard method.All six SLC29A3 exons and their flanking intronic sequences were polymerase chain reaction （PCR）-amplified and the PCR products were subjected to direct sequencing.Results：The patient,an 18-year-old man born to a nonconsanguineous Chinese couple,had more extensive cutaneous lesions,involving both buttocks and knee.In his genomic DNA,we identified a novel homozygous insertion-deletion,c.1269_1270delinsA,in SLC29A3.Both of his parents were carriers of the mutation.Conclusions：We have identified a pathogenic mutation in a Chinese patient with H syndrome.展开更多
目的:研究印记基因SLC22A18(solute carrier family 22,member 18)在乳腺癌中的表达情况及其与乳腺癌侵袭能力的关系。方法:采用Transwell方法评估2种不同恶性程度的乳腺癌细胞株MDA-MB-231(恶性程度高)和MCF-7(恶性程度低)的侵袭转移...目的:研究印记基因SLC22A18(solute carrier family 22,member 18)在乳腺癌中的表达情况及其与乳腺癌侵袭能力的关系。方法:采用Transwell方法评估2种不同恶性程度的乳腺癌细胞株MDA-MB-231(恶性程度高)和MCF-7(恶性程度低)的侵袭转移能力。分别采用实时荧光定量逆转录聚合酶链反应(RT-PCR)和蛋白质印迹法检测SLC22A18的mRNA和蛋白在这2种乳腺癌细胞株中的表达情况。结果:MDA-MB-231细胞株恶性程度高,穿过膜的细胞多,侵袭能力强;MCF-7细胞株恶性程度低,穿过膜的细胞少,侵袭能力弱;SLC22A18在MCF-7中的mRNA和蛋白表达水平高于MDA-MB-231;差异有统计学意义(P<0.01)。结论:印记基因SLC22A18的表达与乳腺癌细胞的侵袭能力相关,该基因有望作为一个抑癌基因抑制乳腺癌的转移。展开更多
目的探究外周血白细胞溶质载体家族45成员4(solute carrier family 45 member 4,SLC45A4)和α血红蛋白稳定蛋白(αhemoglobin stabilizing protein,AHSP)基因甲基化与乳腺癌发病的关系。方法通过差异甲基化分析等方法在GSE51032、GSE104...目的探究外周血白细胞溶质载体家族45成员4(solute carrier family 45 member 4,SLC45A4)和α血红蛋白稳定蛋白(αhemoglobin stabilizing protein,AHSP)基因甲基化与乳腺癌发病的关系。方法通过差异甲基化分析等方法在GSE51032、GSE104942和GSE89093数据集中筛选与乳腺癌发病潜在相关的基因。采用病例对照研究,纳入545例乳腺癌患者和524例非乳腺癌对照作为研究对象。使用MethylTarget靶向测序检测目标基因甲基化并分析其与乳腺癌的关系。结果本研究共筛选出4个基因,控制混杂因素后,SLC45A4和AHSP高甲基化与乳腺癌关系的ORadj分别为0.218(95%CI:0.158~0.299)和0.535(95%CI:0.384~0.741)。在不同亚组中SLC45A4高甲基化与乳腺癌的关联仍有统计学意义,ORadj最高为0.427(95%CI:0.270~0.679),最低为0.153(95%CI:0.085~0.274),而AHSP高甲基化仅在管腔A型(Luminal A)、管腔B型(Luminal B)、雌激素受体阳性(estrogen receptor+,ER+)和≤60岁年龄组中与乳腺癌的关联有统计学意义。异质核糖核酸蛋白C(heterogeneous nuclear ribonucleoproteins C,HNRNPC)甲基化与乳腺癌的关系与公共数据结果相反,ORadj为0.747(95%CI:0.569~0.980),锌指蛋白425(zinc finger protein 425,ZNF425)甲基化与乳腺癌的关联无统计学意义(P=0.158)。结论外周血白细胞SLC45A4和AHSP基因高甲基化可能是乳腺癌发病的保护因素。展开更多
溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11/xCT)作为一种胱氨酸/谷氨酸逆向转运蛋白,参与氨基酸在质膜上的转运,调节细胞铁死亡的发生机制。近年来,越来越多的研究表明,SLC7A11与心血管系统疾病的发生、发展密切...溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11/xCT)作为一种胱氨酸/谷氨酸逆向转运蛋白,参与氨基酸在质膜上的转运,调节细胞铁死亡的发生机制。近年来,越来越多的研究表明,SLC7A11与心血管系统疾病的发生、发展密切相关。本文综述了SLC7A11的结构、功能、调控机制及其在心血管疾病发展中的作用机理,旨在寻找与心血管疾病防治相关的潜在靶点。展开更多
目的:探讨溶质载体家族22成员14(SLC22A14)和精子相关抗原6(SPAG6),在特发性弱精子症患者精子中的表达情况。方法:收集精子库合格捐精者(正常对照组)和特发性弱精子症患者(弱精子症组)各50例精子样本,采用非连续密度梯度离心纯化精子,...目的:探讨溶质载体家族22成员14(SLC22A14)和精子相关抗原6(SPAG6),在特发性弱精子症患者精子中的表达情况。方法:收集精子库合格捐精者(正常对照组)和特发性弱精子症患者(弱精子症组)各50例精子样本,采用非连续密度梯度离心纯化精子,分别用RT-PCR和Western印迹检测SLC22A14、SPAG6 mRNA及蛋白的表达。结果:RT-PCR检测结果显示,弱精子症组的SLC22A14、SPAG6 mRNA表达均显著低于正常对照组(SLC22A14:0.53±0.10 vs 0.77±0.08,t=12.834,P<0.01;SPAG6:0.52±0.10 vs 0.77±0.06,t=13.755,P<0.01),Western印迹检测结果显示,弱精子症组的SLC22A14和SPAG6蛋白表达量同样均显著低于正常对照组,分别为(SLC22A14:0.55±0.10 vs 0.80±0.09,t=12.884,P<0.01;SPAG6:0.56±0.09 vs 0.78±0.09,t=12.257,P<0.01)。结论:在特发性弱精子症患者中SLC22A14和SPAG6表达降低可能是导致弱精子症的重要原因之一。展开更多
文摘Background：H syndrome （OMIM 612391） is a recently described autosomal recessive genodermatosis characterized by indurated hyperpigmented and hypertrichotic skin,as well as other systemic manifestations.Most of the cases occurred in the Middle East areas or nearby countries such as Spain or India.The syndrome is caused by mutations in solute carrier family 29,member 3 （SLC29A3）,the gene encoding equilibrative nucleoside transporter 3.The aim of this study was to identify pathogenic SLC29A 3 mutations in a Chinese patient clinically diagnosed with H syndrome.Methods：Peripheral blood samples were collected from the patient and his parents.Genomic DNA was isolated by the standard method.All six SLC29A3 exons and their flanking intronic sequences were polymerase chain reaction （PCR）-amplified and the PCR products were subjected to direct sequencing.Results：The patient,an 18-year-old man born to a nonconsanguineous Chinese couple,had more extensive cutaneous lesions,involving both buttocks and knee.In his genomic DNA,we identified a novel homozygous insertion-deletion,c.1269_1270delinsA,in SLC29A3.Both of his parents were carriers of the mutation.Conclusions：We have identified a pathogenic mutation in a Chinese patient with H syndrome.
文摘目的:研究印记基因SLC22A18(solute carrier family 22,member 18)在乳腺癌中的表达情况及其与乳腺癌侵袭能力的关系。方法:采用Transwell方法评估2种不同恶性程度的乳腺癌细胞株MDA-MB-231(恶性程度高)和MCF-7(恶性程度低)的侵袭转移能力。分别采用实时荧光定量逆转录聚合酶链反应(RT-PCR)和蛋白质印迹法检测SLC22A18的mRNA和蛋白在这2种乳腺癌细胞株中的表达情况。结果:MDA-MB-231细胞株恶性程度高,穿过膜的细胞多,侵袭能力强;MCF-7细胞株恶性程度低,穿过膜的细胞少,侵袭能力弱;SLC22A18在MCF-7中的mRNA和蛋白表达水平高于MDA-MB-231;差异有统计学意义(P<0.01)。结论:印记基因SLC22A18的表达与乳腺癌细胞的侵袭能力相关,该基因有望作为一个抑癌基因抑制乳腺癌的转移。
文摘溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11/xCT)作为一种胱氨酸/谷氨酸逆向转运蛋白,参与氨基酸在质膜上的转运,调节细胞铁死亡的发生机制。近年来,越来越多的研究表明,SLC7A11与心血管系统疾病的发生、发展密切相关。本文综述了SLC7A11的结构、功能、调控机制及其在心血管疾病发展中的作用机理,旨在寻找与心血管疾病防治相关的潜在靶点。
文摘目的:探讨溶质载体家族22成员14(SLC22A14)和精子相关抗原6(SPAG6),在特发性弱精子症患者精子中的表达情况。方法:收集精子库合格捐精者(正常对照组)和特发性弱精子症患者(弱精子症组)各50例精子样本,采用非连续密度梯度离心纯化精子,分别用RT-PCR和Western印迹检测SLC22A14、SPAG6 mRNA及蛋白的表达。结果:RT-PCR检测结果显示,弱精子症组的SLC22A14、SPAG6 mRNA表达均显著低于正常对照组(SLC22A14:0.53±0.10 vs 0.77±0.08,t=12.834,P<0.01;SPAG6:0.52±0.10 vs 0.77±0.06,t=13.755,P<0.01),Western印迹检测结果显示,弱精子症组的SLC22A14和SPAG6蛋白表达量同样均显著低于正常对照组,分别为(SLC22A14:0.55±0.10 vs 0.80±0.09,t=12.884,P<0.01;SPAG6:0.56±0.09 vs 0.78±0.09,t=12.257,P<0.01)。结论:在特发性弱精子症患者中SLC22A14和SPAG6表达降低可能是导致弱精子症的重要原因之一。
