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EB病毒编码的瘤蛋白潜伏膜蛋白 (LMP1)所介导的活化蛋白 (AP 1)信号转导途径在细胞增殖、分化、转化与凋亡方面发挥着重要作用 .越来越多的证据表明 ,AP 1信号转导通路中上游激酶JNK在鼻咽癌的发生发展过程中起着重要作用 .最近克隆出来的JNK相互作用蛋白 (JIP 1)是一种能抑制JNK核移位的胞浆锚蛋白 .为探讨JIP在LMP1调控AP 1信号通路中的作用机制 ,采用间接免疫荧光法和报告基因法 ,发现JIP通过有效地抑制磷酸化的JNK从胞浆移位入核 ,从而抑制LMP1上调的AP 1活性 .同时 ,JIP导入鼻咽癌细胞中 ,MTT法发现JIP能够明显抑制鼻咽癌细胞的生长 .进一步发现转染JIP后细胞的集落形成率与对照组相比大约降低了 5 3 6 % ,也抑制了细胞 .提示JIP可明显抑制细胞的增殖作用 .进一步采用流式细胞术分析 ,结果发现JIP引起细胞G1/S期细胞阻滞 ,说明JIP是抑制细胞增殖的重要调节子 .进一步采用流式细胞术定量发现 ,转染JIP后细胞的 2 4h凋亡百分率由 1 2 5 %上升到 8 2 5 % ,上升约 6 6倍 ,4 8h由 1 0 4 %上升到 31 4 5 % ,上升约 30倍 .采用激光共聚焦显微镜发现 ,转染JIP后细胞核发生显著变化 ,核质由均匀状态固缩成高凝集状态 ,形成了典型的胞膜体 .提示JIP可有效地促进细胞凋亡 .结果表明 ,JIP可通过抑制活化的JNK?
Epstein-Barr virus coding knoboma protein latent membrane protein (LMP1) -mediated activation of protein (AP1) signal transduction pathway plays an important role in cell proliferation, differentiation, transformation and apoptosis.More and more evidence shows that, The upstream kinase JNK in AP 1 signal transduction pathway plays an important role in the development of nasopharyngeal carcinoma.The recently cloned JNK interacting protein (JIP 1) is a cytoplasmic ankyrin that can inhibit JNK nuclear translocation To explore the mechanism of JIP in regulating AP1 signaling pathway by LMP1, indirect immunofluorescence assay and reporter gene method were used to find out that JIP inhibited the phosphorylation of JNK from the cytoplasm to the nucleus, thereby inhibiting the upregulation of LMP1 AP 1 activity.At the same time, JIP transfected nasopharyngeal carcinoma cells, MTT assay found that JIP can significantly inhibit the growth of nasopharyngeal carcinoma cells.Furthermore, we found that the rate of colony-forming of transfected JIP cells decreased by 53.6% compared with the control group , Also inhibited the cells.It was suggested that JIP can significantly inhibit the proliferation of cells.Further use of flow cytometry analysis, the results showed that JIP caused cell arrest in G1 / S phase, indicating JIP is an important regulator of cell proliferation inhibition. The results of flow cytometry showed that the percentage of apoptotic cells at 24 h after JIP transfection increased from 125% to 82.5%, increased by about 6 6 times and increased from 104% to 31 4 5 at 48 h %, An increase of about 30 times.Using laser confocal microscopy found that after transfection JIP significant changes in the nucleus, the nucleus from the uniform state condensed into a high agglutination state, forming a typical membrane body.It is suggested JIP can effectively promote the cells Apoptosis. The results show that JIP can inhibit the activation of JNK?