干扰hTERT增强光动力疗法诱导宫颈癌细胞凋亡的体外实验  

The exoteric experiment of hTERT interference enhancing the effects of photodynamic therapy in cervical cancer cells by accelerating the apoptosis

作　　者：王颉[1] 黄守国[1] 宗丽菊 张友忠[3] Wang Jie;Huang Shouguo;Zong Liju(Department of Obstetrics and Gynecology,Central South University Xiangya School of Medicine Affiliated Haikou Hospital,Haikou 570208;Department of Obstetrics and Gynecology,Chinese Academy of Medical Sciences & Peking Union Medical College,Beijing 100730)

机构地区：[1]中南大学湘雅医学院附属海口医院妇产科,海口570208 [2]中国医学科学院北京协和医院妇产科,北京100730 [3]山东大学齐鲁医院,济南250012

出　　处：《现代妇产科进展》2021年第11期815-818,共4页Progress in Obstetrics and Gynecology

摘　　要：目的观察干扰hTERT对TMPyP4光动力疗法诱导宫颈癌细胞发生凋亡的作用,并探索其机制。方法筛选hTERT的有效干扰片段,并将其转染至宫颈癌Caski细胞和接受TMPyP4-PDT的Caski细胞。AnnexinV-PE/7-AAD双染试剂盒检测细胞凋亡率,荧光实时定量PCR法、Western blot分别检测Bcl-2、Bax、caspase-3、SP-1基因mRNA和蛋白水平的表达。结果成功筛选sh-4作为有效干扰片段用于后续实验,接受TMPyP4-PDT的Caski细胞稳定转染此片段后,细胞凋亡率明显升高,Bcl-2、SP-1基因在mRNA水平和蛋白水平表达均减少,Bax、caspase-3基因在mRNA水平和蛋白水平表达均增加。结论干扰hTERT基因可以增强光动力疗法(PDT)的效果,可能机制是通过降低靶基因Bcl-2、SP-1表达和增加Bax、caspase-3表达,进而加速细胞凋亡。Objective:To investigate the effects of hTERT interference to the effects of apoptosis induced by meso-5,10,15,20-Tetrakis-(Nmethyl-4-pyridyl)(TMPyP4)-Photodynamic therapy(PDT)in cervical cancer cells and elucidate the mechanisms involved.Methods:The most available hTERT sequence was picked out and transfected into normal Caski cells and those treated by TMPyP4-PDT.The apoptosis was examined by AnnexinV-PE/7-AAD double dye kit.The expressions of mRNA and proteins of Bcl-2,Bax,caspase-3,SP-1 were detected by real time PCR and the Western blot.Results:hTERT gene was successfully silenced by sh-4,which was used in the following experiments.The Caski cells treated by TMPyP4-PDT,were transfected with hTERT-sh4,the expressions of Bcl-2,SP-1 mRNA and protein decreased significantly(P<0.05),meanwhile,the expressions of Bax,caspase-3 mRNA and protein increased.The apoptosis rate increased accordingly.Conclusion:The hTERT interference can enhance the effect of photodynamic therapy in cervical cancer cells by accelerating the apoptosis of cervical cancer cells.

关 键 词：CASKI细胞 sh-hTERT 光动力治疗 凋亡 

分 类 号：R737.33[医药卫生—肿瘤]