沉默FANCD2对人头颈鳞癌SIHN-005A细胞化疗敏感度的影响及其机制

发布时间：2018-11-14 10:17

【摘要】：目的研究FANCD2 sh RNA干扰对人头颈鳞癌(head and neck squamous cell carcinoma,HNSCC)细胞SIHN-005A化疗敏感度的影响,并初步探讨其可能机制。方法应用前期实验获得的FANCD2基因沉默效应稳定的人HNSCC SIHN-005A细胞,以嘌呤霉素持续筛选;Western blot检测沉默效应;CCK-8法检测顺铂作用后细胞增殖抑制率;Hoechst法检测细胞凋亡;Western blot检测Cyclin D1、Bax/Bcl-2、HIF-1α蛋白的表达。结果实验组细胞FANCD2蛋白表达明显下降,FANCD2蛋白沉默效率达60.5%。实验组细胞增殖抑制率明显高于阴性对照组和空白对照组,且具有浓度及时间依赖性,实验组细胞对CDDP的IC50值明显低于阴性对照组和空白对照组。FANCD2基因沉默后实验组细胞凋亡率明显增高。加用CDDP后,Cyclin D1、Bcl-2、HIF-1α蛋白表达水平较加用CDDP前明显下调,Bax蛋白明显增强。结论沉默FANCD2可增加SIHN-005A细胞对化疗药物CDDP的敏感度,其机制可能与FANCD2 sh RNA干扰引起Bax/bcl-2、Cyclin D1、HIF-1α的表达变化有关。
[Abstract]:Objective to study the effect of FANCD2 sh RNA interference on chemotherapeutic sensitivity of human head and neck squamous cell carcinoma (head and neck squamous cell carcinoma,HNSCC) cell line SIHN-005A and its possible mechanism. Methods Human HNSCC SIHN-005A cells with stable effect of FANCD2 gene silencing were used to detect the silencing effect by purine mycin continuous screening; Western blot, the inhibition rate of cell proliferation by CCK-8 assay and apoptosis by Hoechst assay. The expression of HIF-1 伪 protein was detected by Western blot. Results the expression of FANCD2 protein decreased significantly in the experimental group, and the silencing efficiency of FANCD2 protein reached 60.5%. The inhibition rate of cell proliferation in the experimental group was significantly higher than that in the negative control group and the blank control group, and was in a concentration and time dependent manner. The IC50 value of CDDP in the experimental group was significantly lower than that in the negative control group and the blank control group. The apoptosis rate of the experimental group was increased after FANCD2 gene silencing. After adding CDDP, the expression of Bcl-2HIF-1 伪 protein in Cyclin D1 was significantly down-regulated and Bax protein was significantly increased compared with that before CDDP. Conclusion silencing FANCD2 can increase the sensitivity of SIHN-005A cells to the chemotherapeutic drug CDDP, and the mechanism may be related to the change of Bax/bcl-2,Cyclin D1HIF-1 伪 expression induced by FANCD2 sh RNA interference.
【作者单位】：西南医科大学附属医院耳鼻咽喉头颈外科;成都市第一人民医院耳鼻咽喉科;
【基金】：四川省教育厅重点项目(12ZA076) 四川省卫生厅科研项目(120352) 泸州医学院附属医院留学归国人员科研项目([2013]60-2) 四川省科技厅-泸州市科技局-泸州医学院联合科研项目(14JC0182)
【分类号】：R739.91

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