盐酸埃克替尼通过激活蛋白激酶C诱导Tca8113细胞表达iNOS和凋亡

Icotinib Induced the Expression of iNOS and Apoptosis Through the Activation of PKC Pathway in Tca8113 Cells

体外培养Tca8113细胞,随机分为4组,(1)对照组;(2)埃克替尼处理组(20μmo1/L);(3)PKC抑制剂Ro31-8220(3μmol/L)处理组;(4)埃克替尼+Ro31-8220组(20μmo1/L Icotinib,3μmol/L Ro31-8220),不同干预因素处理细胞4h.Western blot检测不同处理组的胞膜内的PKC-α的表达水平,iNOS ELISA检测试剂盒测定细胞的iNOS含量,Griess方法测定Tca8113细胞产生的NO水平,用DNA fragmentation检测Tca8113细胞的凋亡情况.结果表明埃克替尼组DNA fragmentation细胞凋亡及上清液的iNOS,NO较对照组均明显升高;埃克替尼组细胞膜的PKC-α蛋白表达量明显增加.用埃克替尼与PKC的抑制剂Ro31-8220共同处理Tca8113细胞后,胞膜的PKC-α,iNOS的蛋白表达水平及产生的NO能被Ro31-8220显著抑制,且PKC抑制剂Ro31-8220能逆转埃克替尼引起的Tca8113细胞凋亡.埃克替尼能诱导Tca8113细胞iNOS表达和NO生成增加并能诱导Tca8113细胞凋亡;埃克替尼能促进细胞膜内的PKC-α表达增加;埃克替尼诱导的Tca8113细胞iNOS表达和凋亡与PKC有关.

The Tca8113 cells were divided into 4 groups at randomly： control group, Icotinib treated group （20 μmol/ L）, PKC inhibitor Ro31-8220 treated group （3 μmol/L）, Ro31-8220（3 μmol/L）＋Icotinih （20 μmol/L） group for 4 h. The level of PKC-a in membrane and the production of NO and the protein expression of iNOS were examined by western blot, Griess method and iNOS ELISA assay, respectively. Cell apoptosis was examined by DNA fragmentation detection. The results show that Ieotinib stimulated the expression of iNOS and the production of NO in Tca8113 cells. Icotinib also induced apoptosis in Tca8113 cells. The apoptosis induced by Icotinib, the expression of iNOS and NO were inhibited in response to Icotinib combination with PKC inhibitor, Ro31-8220. lcotinib could increase Tca8113 cells to express iNOS and the production of NO and induce apoptosis; Icotinib could upregulate the expression of PKC a in membrane; The expression of iNOS and apoptosis in duced by Icotinib in Tca8113 cells were associated with the activation of PKC pathway.