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前列腺癌LNCaP-AI+F细胞外泌体促进基质细胞WPMY-1功能活化 被引量:4
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作者 范维肖 雷琳 +5 位作者 李蕊 刁艳君 常亮 杨柳 马越云 郝晓柯 《中国肿瘤生物治疗杂志》 CAS CSCD 北大核心 2019年第3期293-298,共6页
目的:探讨前列腺癌外泌体对基质细胞WPMY-1迁移和侵袭能力的影响及其作用机制。方法:超速离心法提取前列腺癌LNCaP-AI+F细胞上清中的外泌体,电镜观察外泌体的典型形态结构,Zetaview检测外泌体的粒径分布,Wb鉴定外泌体标志蛋白及其他相... 目的:探讨前列腺癌外泌体对基质细胞WPMY-1迁移和侵袭能力的影响及其作用机制。方法:超速离心法提取前列腺癌LNCaP-AI+F细胞上清中的外泌体,电镜观察外泌体的典型形态结构,Zetaview检测外泌体的粒径分布,Wb鉴定外泌体标志蛋白及其他相关蛋白。将WPMY-1细胞与前列腺癌外泌体(40μg/ml)共孵育后,激光共聚焦显微镜观察WPMY-1细胞对PKH67标记的外泌体的摄取情况,Transwell实验检测WPMY-1细胞迁移和侵袭能力,qPCR检测IL-8、PDGFB和MMP9等三种肿瘤相关成纤维细胞(cancer-associated fibroblast,CAF)分子表达水平,Wb检测EGFR和ERK1/2蛋白磷酸化水平。结果:电镜下可观察到典型茶托状外泌体结构,外泌体粒径分布集中在100 nm左右,其标志蛋白CD63和ALIX的表达证实了所提取颗粒为外泌体。此外,外泌体还表达EGFR、HER2和SRC等三种与前列腺癌进展相关的蛋白。WPMY-1细胞与外泌体共孵育后,共聚焦显微镜下可看到该细胞摄取大量外泌体,明显促进WPMY-1细胞的迁移和侵袭能力(均P<0.01);与对照组比较,外泌体(40μg/ml)处理后WPMY-1 IL-8、PDGFB及MMP9表达水平增高(P<0.05或P<0.01);且外泌体促进了WPMY-1细胞EGFR和ERK1/2的磷酸化(P<0.01)。结论:前列腺癌细胞可通过外泌体作用于基质细胞WPMY-1,使其高表达多种CAF相关分子,促进EGFR和ERK1/2的磷酸化,增强其迁移和侵袭能力。 展开更多
关键词 前列腺癌 LNCap-AI+F细胞 外泌体 迁移 侵袭 wpmy-1细胞 肿瘤相关成纤维细胞
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Biological effect of human serum collected before and after oral intake of Pygeum africanum on various benign prostate cell cultures 被引量:2
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作者 Stephane Larre Philippe Camparo +6 位作者 Eva Comperat Delphine Boulbes Mohammed Haddoum Sylvain Baulande Pascal Soularue Pierre Costa Olivier Cussenot 《Asian Journal of Andrology》 SCIE CAS CSCD 2012年第3期499-504,I0010,共7页
Pygeum africanum (Tadenan) is a popular phytotherapeutic agent used in the treatment of symptomatic benign prostatic hyperplasia. The active compounds of the drug have not been identified, and determining the plasma... Pygeum africanum (Tadenan) is a popular phytotherapeutic agent used in the treatment of symptomatic benign prostatic hyperplasia. The active compounds of the drug have not been identified, and determining the plasma concentration of the drug is, therefore, not possible. Because there are conflicting results on the efficacy of this drug, we aimed to investigate its effect on prostate cell growth in vitro using human serum collected before and after Pygeum africanum intake. We used primary and organotypic cultures of human prostatic stromal myofibroblast cell line WPMY and prostatic epithelial cell line PNT2. We also used fresh benign prostatic tissue. The serum of a treated man induced decreases in the proliferation of primary cells, organotypic cells and WPMY cells but not PNT2 cells. We also analysed the effect of treated serum on the gene expression profile of WPMY cells. The transcriptome analysis revealed an upregulation of genes involved in multiple tumour suppression pathways and a downregulation of genes involved in inflammation and oxidative-stress pathways. The oral intake of Pygeum africanum resulted in serum levels of active substances that were sufficient to inhibit the proliferation of cultured myofibroblasts prostatic cells. This inhibition was associated with changes in the transcriptome. Asian Journal of Andrology(2012) 14, 499-504; doi:lO.1038/aja.2011.132; published online 26 December 2011 展开更多
关键词 benign prostatic hyperplasia organotypic culture primary culture PNT2 Pygeum africanum TRANSCRIPTOME wpmy
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Qianliening Capsule(前列宁胶囊) Inhibits Human Prostate Cell Growth via Induction of Mitochondrion-Dependent Cell Apoptosis 被引量:3
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作者 洪振丰 林久茂 +4 位作者 钟晓勇 李颖 周建衡 徐伟 彭军 《Chinese Journal of Integrative Medicine》 SCIE CAS 2012年第11期824-830,共7页
Objective: To investigate the molecular mechanisms by which Qianliening Capsule (前列宁胶囊,QC) treats benign prostatic hyperplasia (BPH). Methods: Human prostate stromal cell line WPMY1 was treated with 0, 1, 3... Objective: To investigate the molecular mechanisms by which Qianliening Capsule (前列宁胶囊,QC) treats benign prostatic hyperplasia (BPH). Methods: Human prostate stromal cell line WPMY1 was treated with 0, 1, 3 and 5 mg/mL of QC for 24, 48 and 72 h, respectively, in the presence of 10 ng/mL basic fibroblast growth factor (bFGF). The viability of WPMY1 cells was determined by 3(4,5Dimethylthiazol2yl)2,5diphenyltetrazolium bromide (M'lr) assay. Cell morphology was observed by phasecontrast microscopy. 4',6diamidino2phenylindole (DAPI) staining and fluorescence activated cell sorting (FACS) analysis with AnnexinV/propidium iodide (PI) staining were performed to determine cell apoptosis. The loss of mitochondrial membrane potential was examined by FACS analysis with 5,5',6,6'tetrachloro1 ,l',3,3'tetraethylbenzimidazolylcarbocyadne iodide (JC1) staining. Activation of caspase3 and 9 was evaluated by colorimetric assay. The mRNA and protein expression levels of Bcl2 and Bax were measured by reverse transcription polymerase chain reaction (RTPCR) and Western blotting, respectively. Results: Upon bFGF stimulation, the viability of WPMY1 cells was increased to 122%118% compared with the control cells (P〈0.05). However, treatment with 15 mg/mL of QC for 24, 48 and 72 h decreased the viability of bFGFsUmulated cells to 80%92%, 59%82%, 36%62% compared with the untreated cells (P〈0.05). In addition, QC treatment reduced WPMY1 cell density in a dosedependent manner. Moreover, QC treatment dosedependently induced the loss of plasma membrane asymmetry, the nuclear condensation and fragmentation, collapse of mitochondrial membrane potential, activation of caspase9 and caspase3, and increase of proapoptotic Bax/Bcl2 ratio. Conclusion: Promoting mitochondriondependent apoptosis of prostate stromal cells might be one of the mechanisms by which QC treats BPH. 展开更多
关键词 Qianliening Capsule Chinese medicine benign prostatic hyperplasia apoptosis wpmy-1 cells
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