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目的:研究胆固醇酯转运蛋白(cholesteryl ester transfer protein,CETP)高表达对宫颈癌HeLa细胞放射敏感性的影响。方法:采用脂质体介导法将构建插入全长CETP基因的pcDNA3载体转染人宫颈癌HeLa细胞,获得稳定高表达CETP的HeLa细胞,同时转染pcDNA3空质粒为对照;克隆形成实验检测细胞的存活率;FCM法检测细胞的凋亡率;蛋白质印迹法检测凋亡相关蛋白Bax、Bcl-2及核因子-κB(nuclear factor-kappa B,NF-κB)的表达。结果:X-射线照射后,CETP高表达的HeLa-CETP细胞的放射增敏比SERD0和SERDq分别为1.42和2.09,而未转染细胞与转染空质粒的细胞照射后,细胞存活率差异无统计学意义。FCM结果显示,在X-射线照射后,HeLa-CETP细胞的凋亡率明显高于未转染细胞与转染空质粒的细胞(P<0.05)。蛋白质印迹法检测结果显示,提高CETP的表达可以降低抗细胞凋亡相关蛋白Bcl-2和NF-кB的表达,而增加促细胞凋亡Bax蛋白的表达。结论:导入外源性CETP基因,提高HeLa细胞中CETP表达水平可明显增加HeLa细胞的放射敏感性,其作用机制可能与增加辐射诱导细胞凋亡和改变凋亡相关蛋白的表达有关。
Objective: To investigate the effect of cholesterol ester transfer protein (CETP) overexpression on the radiosensitivity of cervical cancer HeLa cells. Methods: The pcDNA3 vector with the full-length CETP gene was inserted into HeLa cells by liposome-mediated method to obtain stable and highly expressed CETP HeLa cells transfected with pcDNA3 empty plasmid as control; clone formation assay The cell viability was detected by FCM. The expression of Bax, Bcl-2 and NF-κB were detected by Western blotting. Results: After X-ray irradiation, the CETP highly expressed HeLa-CETP cells had higher radiosensitivities than SERD0 and SERDq of 1.42 and 2.09, respectively. However, the cell viability of untransfected and transfected cells was no difference Statistical significance. FCM results showed that the apoptosis rate of HeLa-CETP cells was significantly higher than that of untransfected cells and empty plasmid transfected cells after X-ray irradiation (P <0.05). Western blotting results showed that increasing CETP expression could decrease the expression of anti-apoptosis related proteins Bcl-2 and NF-кB, and increase the expression of pro-apoptotic Bax protein. CONCLUSION: The introduction of exogenous CETP gene and the increase of CETP expression in HeLa cells can significantly increase the radiosensitivity of HeLa cells. The mechanism may be related to increasing radiation-induced apoptosis and altering the expression of apoptosis related proteins.