Guo Yuan-Yuan, Cui Jian, Hou Chun-Mei, Wang Qing-Yang, Wang Jing, Ma Yuan-Fang, Zhang Ji-Yan
Department of Molecular Immunology, Academy of Military Medical Sciences, Beijing 100850, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2011 Feb;19(1):100-4.
This study was purposed to explore the effect of hyperactivation of c-Jun NH(2)-terminal protein kinase (JNK) on the proliferation of B lymphoma cells. The human B lymphoma cell lines Daudi and Raji were chosen as research objects. The expression of JNK protein was determined by Western blot. The subcellular localization of JNK protein was detected by immunofluorescence. The cell cycle was analyzed by flow cytometry. The suppressive effect of JNK inhibitor SP600125 on the proliferation of Daudi and Raji cells was assayed by ATPLite method. The results demonstrated that hyperactivation of JNK has been found in Daudi and Raji cells. Immunofluorescence confirmed the aberrant subcellular localization of JNK protein in Daudi and Raji cells. Cell cycle assay revealed that Daudi and Raji cells underwent G(2)-M arrest in the presence of SP600125. Furthermore, Daudi and Raji cells showed significant increase in sub-G(1) population, an indicator of apoptotic cells, with the treatment of JNK inhibitors. These data suggested that JNK inhibitors suppressed the growth of B lymphoma cells via cell cycle arrest and apoptosis. Daudi and Raji cells treated with different concentrations of JNK selective inhibitor SP600125 showed dose-dependent reduction in the growth of Daudi and Raji cells. It is concluded that hyperactivation of JNK enhance the proliferation of Daudi and Raji cells. The aberrant subcellular localization of JNK protein may facilitate the nuclear accumulation of basal JNK activity, which made JNK to be a potential target to treat human B lymphoma.
本研究旨在探讨c-Jun氨基末端蛋白激酶（JNK）过度激活对B淋巴瘤细胞增殖的影响。选取人B淋巴瘤细胞系Daudi和Raji作为研究对象。采用蛋白质免疫印迹法检测JNK蛋白的表达。通过免疫荧光检测JNK蛋白的亚细胞定位。采用流式细胞术分析细胞周期。采用ATPLite法检测JNK抑制剂SP600125对Daudi和Raji细胞增殖的抑制作用。结果表明，在Daudi和Raji细胞中发现了JNK的过度激活。免疫荧光证实了Daudi和Raji细胞中JNK蛋白的异常亚细胞定位。细胞周期分析显示，在存在SP600125的情况下，Daudi和Raji细胞发生G(2)-M期阻滞。此外，用JNK抑制剂处理后，Daudi和Raji细胞中凋亡细胞的指标亚G(1)期细胞群显著增加。这些数据表明，JNK抑制剂通过细胞周期阻滞和凋亡抑制B淋巴瘤细胞的生长。用不同浓度的JNK选择性抑制剂SP600125处理Daudi和Raji细胞后，Daudi和Raji细胞的生长呈剂量依赖性降低。结论是，JNK的过度激活增强了Daudi和Raji细胞的增殖。JNK蛋白的异常亚细胞定位可能促进基础JNK活性的核积累，这使得JNK成为治疗人类B淋巴瘤的潜在靶点。
