Yang Yang, Dong Hong-Juan, Gao Guang-Xun, Wang Yi-Wei, Gu Hong-Tao, Shu Mi-Mi, Zhu Hua-Feng, Chen Xie-Qun
Department of Hematology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, Shaanxi Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2010 Oct;18(5):1177-80.
This study was purposed to investigate the effect of xbp-1 gene silencing on bortezomib-induced apoptosis in multiple myeloma cell line NCI-H929 (H929). After xbp-1 gene expression was interfered by small hairpin RNA, the cell apoptosis was assayed by flow cytometry with Annexin V-FITC/PI staining, and the expression level of XBP-1 protein was detected by Western blot. The results showed that XBP-1 protein level of H929 cells was inhibited effectively by the PLL3.7 lentiviral vector mediated expression xbp-1 shRNA. The apoptosis rate was significantly higher in xbp-1 shRNA-expressing cells than in untreated control group [(10.13±0.61)% vs (2.5±0.2)%, p<0.05]. After treatment with bortezomib, the apoptosis rate of XBP-1 protein functionally deficient H929 cells was significantly higher than those in vector control group [(45.07±1)% vs (19.53±0.8)%, p<0.05]. It is concluded that xbp-1 gene silencing can significantly enhance the pro-apoptotic activity of bortezomib in multiple myeloma cells.
本研究旨在探讨xbp - 1基因沉默对硼替佐米诱导多发性骨髓瘤细胞系NCI - H929（H929）凋亡的影响。在小发夹RNA干扰xbp - 1基因表达后，采用Annexin V - FITC/PI染色通过流式细胞术检测细胞凋亡，并通过蛋白质免疫印迹法检测XBP - 1蛋白的表达水平。结果显示，PLL3.7慢病毒载体介导的xbp - 1短发夹RNA有效抑制了H929细胞的XBP - 1蛋白水平。表达xbp - 1短发夹RNA的细胞凋亡率显著高于未处理的对照组[（10.13±0.61）%对（2.5±0.2）%，p<0.05]。用硼替佐米处理后，XBP - 1蛋白功能缺陷的H929细胞凋亡率显著高于载体对照组[（45.07±1）%对（19.53±0.8）%，p<0.05]。结论是，xbp - 1基因沉默可显著增强硼替佐米在多发性骨髓瘤细胞中的促凋亡活性。
