组蛋白去乙酰化酶抑制剂MS-275提高宫颈癌SiHa细胞放射敏感性实验研究

【目的】探讨组蛋白去乙酰化酶抑制剂MS-275对人宫颈癌SiHa细胞放射敏感性的影响及其可能的机制。【方法】应用MS-275体外作用于人宫颈癌SiHa细胞,定量PCR方法检测NF-kB基因和PUMA基因mRNA的表达,Western blotting法检测细胞乙酰化组蛋白H4的表达,应用6MV直线加速器,分别以0、2、4、6、8 Gy不同剂量的X线照射细胞,流式细胞仪检测SiHa细胞凋亡率,克隆形成实验检测SiHa细胞的放射敏感性。【结果】人宫颈癌SiHa细胞经MS-275体外作用后,乙酰化组蛋白H4表达增加,并随MS-275剂量的增加而增加,同时细胞PUMA基因mRNA表达随MS-275剂量升高而增加,NF-κB基因mRNA表达随MS-275剂量增加而降低。与对照组相比,MS-275各组细胞凋亡率增高,并随MS-275剂量的增加而升高,SF2值降低。【结论】MS-275体外能提高人宫颈癌SiHa细胞对放射线损伤的敏感性;增加乙酰化组蛋白H4表达,下调NF-κB基因表达和上调PUMA基因表达是其可能的作用机制。

Study on the effect of histone deacetylase inhibitor MS-275 in the enhance of radiosensitivity in human cervical carcinoma cell line SiHa

[Objective]To study the effect of histone deaeetylase inhibitor MS-275 on the radiosensitivity of human cervical carcinoma SiHa cells and its possible mechanism. [Methods]SiHa cells were treated with MS-275 in vitro. Western blotting was used to detect aeetyl level of histone H4, and real time PCR was used to detect mRNA of NF- κB and PUMA genes. Furlhermore, the SiHa cells were irradiated by different dose of X-ray （0, 2, 4, 6, and 8 Gy） by 6 MY linearity accelerator. Flow cytometry was used to detect eell apoptasis rates and colony formation test was used to explore cell radiosensitivity. [ Results ] After treatment with MS-275, the acetyl level of histone H4 increased in SiHa cells in a concentration dependent manner. Meanwhile, the expressions of NF- κ B gone mRNA decreased and PUMA gone mRNA increased in SiHa eells in a concentration dependent manner. After irradiated by X-ray, the apoptotic rate increased in a concentration dependent manner. The SF2 value of MS-275 group was depressed. [ Conclusion ] MS-275 could enhance radiosensitivity in SiHa cells. Increasing acetyl level of histone, down-regulate NF- κB and up-regulate PUMA gone expressions may be its mechanism.