真核载体介导STAT3基因沉默诱导CNE2细胞凋亡的研究  

Inhibition of STAT3 gene expression by eukaryotic vector inducing apoptosis in CNE2 cell line

作　　者：刘伟[1] 王涛[1] 陈叶珊[1] 姚杰[1] 胡建莉[1] 

机构地区：[1]华中科技大学附属协和医院肿瘤中心,湖北武汉430022

出　　处：《中华肿瘤防治杂志》2009年第16期1217-1220,共4页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的:研究真核载体介导的siRNA对人鼻咽癌CNE2细胞凋亡的影响。方法:真核质粒介导的针对STAT3基因siRNA转染人鼻咽癌细胞CNE2;RT-PCR、蛋白质印迹法、激光共聚焦显微镜、流式细胞术分别检测STAT3mRNA表达、STAT3、CyclinD1、Bcl-2和Bax蛋白表达以及细胞凋亡形态学改变、细胞周期和细胞凋亡。结果:转染72h后,与未转染组和阴性对照组相比,STAT3基因沉默组CNE2细胞中STAT3mRNA(P=0.000)和蛋白表达显著下降(P=0.000);Cyc-linD1和Bcl-2蛋白表达明显减少(P=0.000),Bax蛋白表达显著增多(P=0.000);Hoechst33258荧光染色可见核浓缩等典型细胞凋亡特征;细胞周期G1期比例明显增多〔分别为(80.09±2.37)%、(57.98±4.19)%和(61.85±2.31)%〕,P=0.000;凋亡率显著升高〔分别为(16.4±3.78)%、(3.20±1.74)%和(3.42±0.57)%〕,P=0.000。结论:STAT3基因沉默能有效地抑制鼻咽癌CNE2细胞中的STAT3表达,并通过调节CyclinD1、Bcl-2和Bax蛋白表达使鼻咽癌细胞阻滞于G1期,最终引致细胞凋亡。OBJECTIVE： To investigate the influence of STAT3-specific RNA interference on apoptosis of human nasopharyngeal carcinoma cell line CNE2. METHODS： pGenesil- STAT3-siRNA was synthesized and transfected into CNE2 cells by liposome. The expression of STAT3 gene in CNE2 cells was detected by RT-PCR, and the expressions of STAT3, Cyclin D1, Bcl-2 and bax proteins were detected by Western-blot. The cell apoptosis and cell cycle were detected by Hoechst 33258 and flow cytometry （FCM） in CNE2 cells. RESULTS： After 72 h of transfection with siRNA, the mRNA （P= 0. 000） and protein levels （P= 0. 000） of STAT3 in CNE2 cells were significantly lower than those of the untreated and scrambled group, accompanied by a decrease of the expressions of Cyclin D1 and Bcl-2 proteins （P= 0. 000） and an augmentation of Bax protein expression （P = 0. 000）. Marked morphological changes of apoptosis were ob- served by Hoechst 33258 staining after transfected. The cell cycle progression was blocked at G1 stage （（80.09±2.37） %, （57.98± 4.19） %, （61.85± 2.31 ） %, P = 0. 0003, and the apoptosis rates increased [（16.4±3.78）%, （3.20± 1.74）%, （3.42±0.57）%, P= 0. 000 ] in the siRNA treated group. CONCLUSION： STAT3-siRNA can effectively inhibit the expression of STAT3, induce cell apoptosis and inhibit cell proliferation by the regulation of the expressions of Bax, Cyclin D1 and Bcl-2 proteins in CNE2 cells.

关 键 词：鼻咽肿瘤 STAT3 RNA干扰 细胞凋亡 

分 类 号：R739.63[医药卫生—肿瘤]