目的探讨谷氨酸-脯氨酸-亮氨酸富集蛋白1(proline-,glutamic acid-,and leucine-rich protein 1,PELP1)在胃癌中的生物学功能及临床意义。方法通过生物信息学方法分析PELP1在胃癌中的表达,并在胃癌细胞系和胃癌临床样本中予以验证,Kapla...目的探讨谷氨酸-脯氨酸-亮氨酸富集蛋白1(proline-,glutamic acid-,and leucine-rich protein 1,PELP1)在胃癌中的生物学功能及临床意义。方法通过生物信息学方法分析PELP1在胃癌中的表达,并在胃癌细胞系和胃癌临床样本中予以验证,Kaplan-Meier生存分析探讨PELP1表达与胃癌患者生存预后的关系。以siRNA敲降胃癌细胞AGS中的PELP1表达,以CCK-8实验、平板克隆形成实验、EdU细胞增殖实验检测细胞增殖活性;流式细胞术测定细胞周期;细胞划痕实验、Transwell小室检测细胞迁移、侵袭能力。Western blot和qRT-PCR检测敲降PELP1表达的AGS细胞中Src-Erk通路信号分子表达情况。结果PELP1在胃癌组织中表达相对较高,生存分析显示PELP1高表达的患者生存期更短;敲降PELP1表达后的胃癌细胞增殖、迁移、侵袭等恶性生物学行为均受到抑制;敲降PELP1表达的AGS细胞,其Src-Erk通路关键信号分子c-Src、PI3K和Erk的表达均降低。结论PELP1在胃癌的发生发展中起致瘤作用,可能是胃癌潜在的治疗靶点。展开更多
目的探讨脯氨酸-谷氨酸-亮氨酸富集蛋白1(Proline,glutamate and leucine rich protein 1,PELP1)表达与胃癌演进及预后的关系及意义。方法采用免疫组化检测39例正常胃组织、48例低级别不典型增生胃组织、28例高级别不典型增生胃组织、7...目的探讨脯氨酸-谷氨酸-亮氨酸富集蛋白1(Proline,glutamate and leucine rich protein 1,PELP1)表达与胃癌演进及预后的关系及意义。方法采用免疫组化检测39例正常胃组织、48例低级别不典型增生胃组织、28例高级别不典型增生胃组织、77例胃癌组织中PELP1蛋白表达水平,观察胃癌不同演进阶段PELP1表达变化;分析胃癌患者PELP1表达水平与临床病理参数关系;通过在线癌症生存分析工具Kaplan-Meier Plotter分析胃癌PELP1表达与患者总生存期、首次进展前生存期和进展后生存期之间的关系。结果胃癌演进过程中,PELP1表达水平逐渐升高,差异具有统计学意义(P<0.05);胃癌组织中PELP1表达与淋巴结转移、低肿瘤分化、高临床分期有关(P<0.05);PELP1高表达组患者总生存期、首次进展前生存期和进展后生存期均显著低于PELP1低表达组患者(P<0.05)。结论高表达PELP1参与胃癌演进并且提示患者预后不良。展开更多
目的建立针对PELP1基因启动子区CpG岛的甲基化特异性PCR(Methylation specific PCR,MSP)检测方法,分析乳腺癌细胞中PELP1基因表达与启动区CpG岛甲基化的相关性。方法设计针对PELP1基因启动子区的MSP引物组,以甲基化和非甲基化DNA为模板...目的建立针对PELP1基因启动子区CpG岛的甲基化特异性PCR(Methylation specific PCR,MSP)检测方法,分析乳腺癌细胞中PELP1基因表达与启动区CpG岛甲基化的相关性。方法设计针对PELP1基因启动子区的MSP引物组,以甲基化和非甲基化DNA为模板验证MSP引物的特异性,建立针对PELP1基因启动子区的MSP检测方法。以DNA甲基转移酶抑制剂5'-氮杂-脱氧胞苷磷酸(5'-Aza-d C)分别处理MCF-7乳腺癌细胞、MCF-10正常乳腺导管上皮细胞,采用MSP检测PELP1基因启动子区甲基化状态变化,采用Western blot检测蛋白表达。结果针对PELP1基因启动子区设计的MSP引物组特异性良好,甲基化特异性引物仅在甲基化DNA模板获得阳性扩增条带,非甲基化引物仅在非甲基化DNA模板获得阳性条带。MCF-7乳腺癌细胞中PELP1基因启动子区呈非甲基化状态,MCF-10正常乳腺导管上皮细胞中PELP1基因启动子区呈甲基化状态,MCF-10细胞中PELP1蛋白表达水平为MCF-7细胞的1/16(P<0.05)。采用5'-Aza-dC去除MCF-10细胞PELP1基因启动子区甲基化后,PELP1蛋白表达水平上升了9.7倍(P<0.05)。结论所建立的PELP1基因启动子区MSP检测方法特异性良好,去甲基化可能是导致乳腺癌细胞中PELP1基因过表达的重要机制。展开更多
Proline-, glutamic acid-, and leucine-rich protein 1 （PELP1） is a scaffolding protein involved in both genomic and nongenomic estrogen signal transduction pathways. To date, the role of PELP1 protein has yet to be c...Proline-, glutamic acid-, and leucine-rich protein 1 （PELP1） is a scaffolding protein involved in both genomic and nongenomic estrogen signal transduction pathways. To date, the role of PELP1 protein has yet to be characterized in human sperm and has not been associated with sperm parameters. To confirm the presence of PELP1 in human sperm, fresh semen samples were obtained from 178 donors. The study was designed to establish both mRNA and protein presence, and protein cellular localization. Additionally, the number of PELP1-positive spermatozoa was analyzed in men with normal and abnormal semen parameters. Sperm parameters were assessed according to the World Health Organization （WHO） 2010 standards. The presence of PELP1 in spermatozoa was investigated using four precise, independent techniques. The qualitative presence of transcripts and protein was assessed using reverse transcription-polymerase chain reaction （RT-PCR） and western blot protocols, respectively. The cellular localization of PELP1 was investigated by immunocytochemistry. Quantitative analysis of PELP1-positive cells was done by flow cytometry. PELP1 mRNA and protein was confirmed in spermatozoa. Immunocytochemical analysis identified the presence of PELP1 in the midpieces of human sperm irrespective of sperm parameters. Becton Dickinson fluorescence-activated cell sorting （FACSCalibur^Tm） analysis revealed a significantly lower number of PELP1-positive cells in males with normal semen parameters versus abnormal samples （42.78% ± 11.77% vs 61.05% ± 21.70%, respectively; P = 0.014）. The assessment of PELP1 may be a time-saving method used to obtain information about sperm quality. The results of our study suggest that PEPL1 may be utilized as an indicator of sperm quality; thereby, PELP1 may be an additional biomarker useful in the evaluation of male infertility.展开更多
[目的]评价SH3结构域谷氨酸富集样蛋白3(SH3BGRL3)在非小细胞肺癌中的表达及其与临床病理特征的相关性。[方法]选取2014年10月至2017年12月确诊并接受手术治疗的46例非小细胞肺癌患者的癌组织标本以及距离癌组织5cm以上的癌旁组织,免疫...[目的]评价SH3结构域谷氨酸富集样蛋白3(SH3BGRL3)在非小细胞肺癌中的表达及其与临床病理特征的相关性。[方法]选取2014年10月至2017年12月确诊并接受手术治疗的46例非小细胞肺癌患者的癌组织标本以及距离癌组织5cm以上的癌旁组织,免疫组化染色检测SH3BGRL3相对表达量,荧光定量RT-PCR检测SH3BGRL3 m RNA表达。[结果]癌组织中SH3BGRL3相对表达量、SH3BGRL3 mRNA表达高于癌旁组织(P<0.05)。肿瘤体积≥5cm3、侵及浆膜、病理分期Ⅲ~Ⅳ期、有淋巴结转移患者SH3BGRL3、SH3BGRL3 m RNA相对表达量高于肿瘤体积<5cm3、未及浆膜、病理分期Ⅰ~Ⅱ期、无淋巴结转移患者(P<0.05)。病理分期、SH3BGRL3、SH3BGRL3 mRNA表达是影响非小细胞肺癌患者预后的主要危险因素。[结论]SH3BGRL3在非小细胞肺癌中呈现异常高表达,且与肿瘤体积、浸润程度、病理分期、淋巴结转移等临床病理参数具有一定相关性。SH3BGRL3、SH3BGRL3 m RNA表达是影响非小细胞肺癌患者预后的危险因素。展开更多
文摘目的探讨脯氨酸-谷氨酸-亮氨酸富集蛋白1(Proline,glutamate and leucine rich protein 1,PELP1)表达与胃癌演进及预后的关系及意义。方法采用免疫组化检测39例正常胃组织、48例低级别不典型增生胃组织、28例高级别不典型增生胃组织、77例胃癌组织中PELP1蛋白表达水平,观察胃癌不同演进阶段PELP1表达变化;分析胃癌患者PELP1表达水平与临床病理参数关系;通过在线癌症生存分析工具Kaplan-Meier Plotter分析胃癌PELP1表达与患者总生存期、首次进展前生存期和进展后生存期之间的关系。结果胃癌演进过程中,PELP1表达水平逐渐升高,差异具有统计学意义(P<0.05);胃癌组织中PELP1表达与淋巴结转移、低肿瘤分化、高临床分期有关(P<0.05);PELP1高表达组患者总生存期、首次进展前生存期和进展后生存期均显著低于PELP1低表达组患者(P<0.05)。结论高表达PELP1参与胃癌演进并且提示患者预后不良。
文摘目的建立针对PELP1基因启动子区CpG岛的甲基化特异性PCR(Methylation specific PCR,MSP)检测方法,分析乳腺癌细胞中PELP1基因表达与启动区CpG岛甲基化的相关性。方法设计针对PELP1基因启动子区的MSP引物组,以甲基化和非甲基化DNA为模板验证MSP引物的特异性,建立针对PELP1基因启动子区的MSP检测方法。以DNA甲基转移酶抑制剂5'-氮杂-脱氧胞苷磷酸(5'-Aza-d C)分别处理MCF-7乳腺癌细胞、MCF-10正常乳腺导管上皮细胞,采用MSP检测PELP1基因启动子区甲基化状态变化,采用Western blot检测蛋白表达。结果针对PELP1基因启动子区设计的MSP引物组特异性良好,甲基化特异性引物仅在甲基化DNA模板获得阳性扩增条带,非甲基化引物仅在非甲基化DNA模板获得阳性条带。MCF-7乳腺癌细胞中PELP1基因启动子区呈非甲基化状态,MCF-10正常乳腺导管上皮细胞中PELP1基因启动子区呈甲基化状态,MCF-10细胞中PELP1蛋白表达水平为MCF-7细胞的1/16(P<0.05)。采用5'-Aza-dC去除MCF-10细胞PELP1基因启动子区甲基化后,PELP1蛋白表达水平上升了9.7倍(P<0.05)。结论所建立的PELP1基因启动子区MSP检测方法特异性良好,去甲基化可能是导致乳腺癌细胞中PELP1基因过表达的重要机制。
文摘Proline-, glutamic acid-, and leucine-rich protein 1 （PELP1） is a scaffolding protein involved in both genomic and nongenomic estrogen signal transduction pathways. To date, the role of PELP1 protein has yet to be characterized in human sperm and has not been associated with sperm parameters. To confirm the presence of PELP1 in human sperm, fresh semen samples were obtained from 178 donors. The study was designed to establish both mRNA and protein presence, and protein cellular localization. Additionally, the number of PELP1-positive spermatozoa was analyzed in men with normal and abnormal semen parameters. Sperm parameters were assessed according to the World Health Organization （WHO） 2010 standards. The presence of PELP1 in spermatozoa was investigated using four precise, independent techniques. The qualitative presence of transcripts and protein was assessed using reverse transcription-polymerase chain reaction （RT-PCR） and western blot protocols, respectively. The cellular localization of PELP1 was investigated by immunocytochemistry. Quantitative analysis of PELP1-positive cells was done by flow cytometry. PELP1 mRNA and protein was confirmed in spermatozoa. Immunocytochemical analysis identified the presence of PELP1 in the midpieces of human sperm irrespective of sperm parameters. Becton Dickinson fluorescence-activated cell sorting （FACSCalibur^Tm） analysis revealed a significantly lower number of PELP1-positive cells in males with normal semen parameters versus abnormal samples （42.78% ± 11.77% vs 61.05% ± 21.70%, respectively; P = 0.014）. The assessment of PELP1 may be a time-saving method used to obtain information about sperm quality. The results of our study suggest that PEPL1 may be utilized as an indicator of sperm quality; thereby, PELP1 may be an additional biomarker useful in the evaluation of male infertility.
文摘[目的]评价SH3结构域谷氨酸富集样蛋白3(SH3BGRL3)在非小细胞肺癌中的表达及其与临床病理特征的相关性。[方法]选取2014年10月至2017年12月确诊并接受手术治疗的46例非小细胞肺癌患者的癌组织标本以及距离癌组织5cm以上的癌旁组织,免疫组化染色检测SH3BGRL3相对表达量,荧光定量RT-PCR检测SH3BGRL3 m RNA表达。[结果]癌组织中SH3BGRL3相对表达量、SH3BGRL3 mRNA表达高于癌旁组织(P<0.05)。肿瘤体积≥5cm3、侵及浆膜、病理分期Ⅲ~Ⅳ期、有淋巴结转移患者SH3BGRL3、SH3BGRL3 m RNA相对表达量高于肿瘤体积<5cm3、未及浆膜、病理分期Ⅰ~Ⅱ期、无淋巴结转移患者(P<0.05)。病理分期、SH3BGRL3、SH3BGRL3 mRNA表达是影响非小细胞肺癌患者预后的主要危险因素。[结论]SH3BGRL3在非小细胞肺癌中呈现异常高表达,且与肿瘤体积、浸润程度、病理分期、淋巴结转移等临床病理参数具有一定相关性。SH3BGRL3、SH3BGRL3 m RNA表达是影响非小细胞肺癌患者预后的危险因素。
