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Effects of protein kinase C and motigen-activated protein kinase kinase/extracellular regulated protein kinases signaling pathway on mRNA level of inducible nitric oxide synthase in Tca8113 cells



Effects of protein kinase C and motigen-activated protein kinase kinase/extracellular regulated protein kinases signaling pathway on mRNA level of inducible nitric oxide synthase in Tca8113 cells



Hua Xi Kou Qiang Yi Xue Za Zhi 36(2): 133-139



目的 在舌鳞癌细胞Tca8113中,探索与细胞增殖密切相关的诱导型一氧化氮合酶(NOS-2)的表达调控机制。方法 采用RNAi技术沉默Tca8113细胞中NOS-2、蛋白激酶C(PKC)-α、PKC-β和PKC-δ的基因;Griess Reagent法检测NOS-2基因沉默后一氧化氮(NO)生成量;CCK8法测定细胞增殖活性;实时定量荧光聚合酶链反应(q-
PCR)技术检测各种方法处理后NOS-2、PKC-α、PKC-β和PKC-δ的基因表达;Western blotting技术测定丙二醇甲醚醋酸酯(PMA)作用细胞后的细胞外调节蛋白激酶(ERK)1/2磷酸化程度。结果 利用NOS-2的siRNA处理后,Tca8113细胞的增殖能力明显降低(P<0.01);PKC的活性与NOS-2的基因表达成负相关(P<0.05);PKC亚型PKC-
α、PKC-β和PKC-δ共同参与NOS-2的基因调控(P<0.01);丝裂原活化蛋白激酶激酶(MEK)/ERK通路负调控NOS-2的基因表达(P<0.05);PKC通过MEK/ERK通路负调控NOS-2的基因表达(P<0.05)。结论 在Tca8113细胞中,PKC通过MEK/ERK通路负调控与细胞增殖相关的NOS-2的基因表达。.

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Accession: 060566416

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PMID: 29779272


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