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TAMs激活Src-RhoA通路上调免疫抑制因子表达及促进直肠癌细胞增殖和EMT 被引量:5
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作者 张静 胡哲 居红格 《免疫学杂志》 CAS CSCD 北大核心 2021年第7期596-603,共8页
目的探讨肿瘤相关巨噬细胞(tumor-associated macrophages,TAMs)对直肠癌增殖和EMT的影响及其作用机制。方法用佛波酯(PMA)及白介素4(interleukin 4,IL-4)诱导人单核巨噬细胞(THP1),使其分化为M2型TAMs,采用流式细胞仪检测CD163和CD16... 目的探讨肿瘤相关巨噬细胞(tumor-associated macrophages,TAMs)对直肠癌增殖和EMT的影响及其作用机制。方法用佛波酯(PMA)及白介素4(interleukin 4,IL-4)诱导人单核巨噬细胞(THP1),使其分化为M2型TAMs,采用流式细胞仪检测CD163和CD16的阳性表达率;TAMs与SW837细胞共培养48 h,通过CCK-8法检测SW837细胞活力,流式细胞仪检测细胞凋亡率,RT-qPCR检测IL-10和TGF-β1的表达量,Western blot分析E-cadherin、N-cadherin、IL-10、TGF-β1蛋白的表达以及Src、RhoA蛋白的磷酸化水平;Src通路抑制剂SU6656作用细胞后,分别用CCK-8和流式细胞仪检测SW837细胞活力和细胞总死亡率,Western blot检测细胞中IL-10、TGF-β1、E-cadherin和N-cadherin的表达量。结果THP1分化诱导为TAMs后,CD163的阳性表达率明显高于CD16的阳性表达率(P<0.01);和SW837、SW837+THP1+PMA、SW837+THP1+IL-4细胞相比,SW837与TAMs共培养后使SW837细胞活力明显升高(P<0.05),SW837细胞总死亡率明显降低(P<0.01),N-cadherin表达量明显升高,Ecadherin表达量明显降低,IL-10和TGF-β1蛋白表达量明显升高;TAMs激活直肠癌SW837细胞中Src-RhoA通路,使Src、RhoA蛋白的磷酸化水平明显升高;和SW837+TAMs细胞相比,SW837+TAMs+SU6656细胞活力明显降低,细胞总死亡率明显升高,E-cadherin蛋白表达量明显升高,N-cadherin蛋白表达量明显降低,IL-10和TGF-β1蛋白表达量明显降低。结论肿瘤相关巨噬细胞通过激活Src-RhoA通路产生免疫抑制因子、促进了直肠癌的增殖和EMT。 展开更多
关键词 肿瘤相关巨噬细胞 src-rhoa 免疫抑制因子 直肠癌
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幽门螺杆菌对人胃癌细胞SGC-7901中SHP-2及细胞骨架的影响 被引量:3
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作者 张尤历 陆芬英 +3 位作者 王文兵 张宇川 刘勇攀 吴莺 《世界华人消化杂志》 CAS 北大核心 2008年第26期2916-2921,共6页
目的:研究幽门螺杆菌(Helicobacter pylori,H pylori)刺激人胃癌细胞SGC-7901后,对细胞内SHP-2表达及细胞骨架的影响.方法:将临床分离的胃癌H pylori菌体裂解,以超声提取物刺激SGC-7901细胞1h,逆转录聚合酶链式反应(RT-PCR)半定量法测... 目的:研究幽门螺杆菌(Helicobacter pylori,H pylori)刺激人胃癌细胞SGC-7901后,对细胞内SHP-2表达及细胞骨架的影响.方法:将临床分离的胃癌H pylori菌体裂解,以超声提取物刺激SGC-7901细胞1h,逆转录聚合酶链式反应(RT-PCR)半定量法测定刺激前后SHP-2表达量的变化;并将PCR产物进行克隆和测序.将不同疾病(胃炎,胃溃疡,胃癌)的菌株H pylori超声提取物刺激SGC-7901细胞10min,采用phalloidin对细胞内骨架进行荧光染色,研究其对细胞骨架的影响.将野生型和突变型RhoA质粒转入细胞后,再以胃癌H pylori超声提取物刺激,进一步观察细胞内骨架的变化.结果:胃癌H pylori菌体超声提取物刺激SGC-7901细胞1h后,细胞内SHP-2表达量没有明显变化.胃炎,胃溃疡,胃癌H pylori菌体超声提取物刺激人胃癌细胞SGC-7901后,与对照组相比,细胞内骨架增多(65.4±0.510,63.37±0.475,64.53±0.522vs20.34±1.376,均P<0.05),但这三种菌株间相比无统计学意义.转染入野生型RhoA质粒的细胞经胃癌H pylori菌体超声提取物刺激后,细胞内骨架较未转染者增多(72.99±1.818vs61.78±1.288,P<0.05),而转染入无活性突变型RhoA质粒的细胞经刺激后,细胞内骨架与未转染者无明显差别.结论:胃癌H pylori超声提取物未能引起细胞内SHP-2mRNA表达量的改变;胃炎,胃溃疡,胃癌H pylori菌体超声提取物能够增加细胞内骨架的形成,其作用与H pylori菌种无关,RhoA活性的增强是其中的一个重要中间环节. 展开更多
关键词 幽门螺杆菌 胃癌细胞 SHP-2 细胞骨架 RHOA
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Participation of Morphofunctional Zones in Aging Processes
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作者 T. M. Yavisheva S. D. Shcherbakov 《Advances in Aging Research》 2014年第2期72-78,共7页
There are morphofunctional zones in organism tissues, where proliferation and differentiation processes occur. Daughter cells are differentiated in the electric field excited by 12 mother and daughter cell pairs, whic... There are morphofunctional zones in organism tissues, where proliferation and differentiation processes occur. Daughter cells are differentiated in the electric field excited by 12 mother and daughter cell pairs, which turned out at cambial cell division. With aging, the cambial cell number is reduced to 7, which is close to threshold level (6 cells), at which the differentiation of daughter cells is absent. The depression of cambial cell number with aging is connected with the work of another morphofunctional zone—the hypothalamus, which is the major center of vegetative regulation and initially has very high RhoA activity, which has been established in embryogenesis. Estrogens, influencing over the hypothalamus and activating Src kinase in its nuclei, reduce level of RhoA activity, including SCN, responsible for many biorhythms of an organism. As a result, the hyperestrogenemia and therefore a connective tissue at first occur. Then there happens a hypoestrogenemia that leads to sharp falling of proliferative activity of cells, causing the depression of cambial cell number and possibility of a malignant tumor development. Along with this, there are the deep lesions of hormone regulation, leading to some lethal diseases. Thus, the RhoA increasing in hypothalamus and especially in SCN circadian rhythm can counteract the Src kinase intensifying and prevent the processes connected with this. 展开更多
关键词 Morphofunctional ZONES HYPOTHALAMUS RHOA Protein SRC KINASE Aging
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