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小干扰RNA介导的RIP1基因敲低增强人口腔鳞状细胞癌对奥沙利铂的敏感性

[Small interfering RNA-mediated RIP1 knockdown enhances L-OHP sensitivity of human oral squamous carcinoma cells].

作者信息

Xu Jincheng, Huang Yingying, Li Yang, Pu Longjian, Xia Fei, Jiang Chenchen, Liu Hao, Jiang Zhiwen

机构信息

Department of Stomatology, First Affiliated Hospital of Bengbu Medical College, Bengbu, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2013 Jul;33(7):1004-7.

Abstract

OBJECTIVE

To investigate the effect of small interfering RNA-mediated receptor-interacting protein kinase 1 (RIP1) knockdown on the sensitivity of human oral squamous carcinoma cells to to oxaliplatin (L-OHP)-induced apoptosis and explore a new target for clinical treatment of oral squamous carcinoma.

METHODS

The viability of human oral squamous carcinoma cell line KB exposed to different concentrations (0, 0.25, 0.5, 1, 2, 4 µmol/L) of L-OHP were detected by MTT assay. PI/Annexin V staining was used to observe cell apoptosis in naive KB cells, cell and transfected with pSH1Si-RIP1 or with the empty plasmid. Western blotting was used to detect RIP1 expression in KB cells exposed to L-OHP and in cells transfected with pSH1Si-RIP1.

RESULTS

Exposure to L-OHP (1µmol/L) for 24, 48, 72 h resulted in KB cell survival rates of 67.66%, 55.17%, and 41.34%, respectively, but the cell apoptosis rate was only 9.6% following a 24-h exposure. KB cells transfected with pSH1Si-RIP1 showed an apoptotic rate of 9.4%, which increased to 29.1% following L-OHP exposure. RIP1 expression was first up-regulated and then down-regulated in KB cells treated with L-OHP, and was significantly reduced after cell transfection with pSH1Si-RIP1.

CONCLUSION

Suppression of RIP1 expression increases the apoptotic rate of human oral squamous carcinoma cells, suggesting the potential of RIP1 as a new candidate target for clinical treatment of oral squamous carcinoma.

摘要

目的

研究小干扰RNA介导的受体相互作用蛋白激酶1(RIP1)基因沉默对人口腔鳞状细胞癌细胞对奥沙利铂(L-OHP)诱导凋亡敏感性的影响,探索口腔鳞状细胞癌临床治疗新靶点。

方法

采用MTT法检测人舌鳞状细胞癌细胞系KB在不同浓度(0、0.25、0.5、1、2、4μmol/L)奥沙利铂作用下的细胞活力。采用PI/Annexin V染色观察未处理的KB细胞、转染pSH1Si-RIP1或空质粒的细胞凋亡情况。采用蛋白质免疫印迹法检测奥沙利铂作用下的KB细胞及转染pSH1Si-RIP1的细胞中RIP1的表达。

结果

奥沙利铂(1μmol/L)作用24、48、72小时后,KB细胞存活率分别为67.66%、55.17%和41.34%,但24小时作用后细胞凋亡率仅为9.6%。转染pSH1Si-RIP1的KB细胞凋亡率为9.4%,奥沙利铂作用后升至29.1%。奥沙利铂处理的KB细胞中RIP1表达先上调后下调,转染pSH1Si-RIP1后RIP1表达显著降低。

结论

抑制RIP1表达可增加人口腔鳞状细胞癌细胞凋亡率,提示RIP1有望成为口腔鳞状细胞癌临床治疗新靶点。

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