文献类型：期刊文献

中文题名：茶黄素对人宫颈癌细胞株HeLa顺铂敏感性的影响及其机制

英文题名：Effects of theaflavins on the sensitivity of human cervical cancer HeLa cells to cisplatin

作者：刘丽萍 吴宏文 王胜

第一作者：刘丽萍

机构：[1]贵州理工学院食品药品制造工程学院;[2]南昌大学第一附属医院;[3]贵州医科大学附属医院

第一机构：贵州理工学院食品药品制造工程学院

年份：2018

卷号：58

期号：19

起止页码：5-8

中文期刊名：山东医药

外文期刊名：Shandong Medical Journal

收录：CSTPCD

基金：贵州省科技计划项目(黔科合SY字[2015]345号)

语种：中文

中文关键词：茶黄素;宫颈癌;顺铂;药物疗法;细胞凋亡;核因子κB

外文关键词：theaflavins;cervical carcinoma;cisplatin;drug therapy;apoptosis;nuclear factor Kb

摘要：目的探讨茶黄素对人宫颈癌细胞株HeLa顺铂敏感性的影响,并探讨其相关机制。方法 (1)取对数生长期HeLa细胞接种于96孔板,设观察1~9组和对照1~9组,每组3个复孔。对照1~9组细胞分别加入0、5、10、15、20、25、30、35、40、50μg/mL顺铂,继续培养24 h。观察1~9组先加入10μg/mL的茶黄素,再分别加入0、5、10、15、20、25、30、35、40、50μg/mL顺铂,继续培养24 h。采用MTT法检测各组OD490,计算各组细胞存活率,然后计算顺铂对观察组和对照组HeLa细胞的半数抑制浓度(IC50)。(2)取对数生长期HeLa细胞接种于96孔板。设对照组、顺铂组、茶黄素组和联合组,每组3个复孔。对照组细胞不加药,顺铂组加入10μg/mL的顺铂,茶黄素组加入10μg/mL的茶黄素,联合组加入10μg/mL的顺铂和10μg/mL茶黄素,继续培养24 h后,采用Western blotting法检测凋亡相关蛋白Cleaved Caspase 3、Cleaved Caspase 9、PARP和NF-κB信号通路相关蛋白p-IκBα、p-NF-κB和pAkt。结果 (1)对照1~9组HeLa细胞存活率分别为102.54%±4.21%、98.32%±2.46%、96.11%±3.64%、90.36%±3.91%、86.29%±3.17%、81.46%±3.61%、78.92%±4.18%、70.63%±4.25%、63.28%±4.21%,顺铂对HeLa细胞的IC50为25μg/mL。观察1~9组HeLa细胞存活率分别为102.54%±4.21%、80.21%±3.82%、78.92%±4.24%、77.93%±3.95%、68.26%±3.24%、50.36%±4.29%、45.32%±4.74%、40.25%±3.21%、35.37%±3.16%,顺铂对HeLa细胞的IC50为10μg/mL。(2)顺铂组、茶黄素组、联合组细胞Cleaved Caspase 3、Cleaved Caspase 9、PARP相对表达量均高于对照组(P均<0.05),联合组细胞Cleaved Caspase 3、Cleaved Caspase 9、PARP相对表达量均高于顺铂组、茶黄素组(P均<0.05)。顺铂组、对照组细胞p-IκBα、p-NF-κB和p-Akt相对表达量相比P>0.05,茶黄素组细胞p-IκBα、p-NF-κB和p-Akt相对表达量高于对照组(P均<0.05),联合组细胞pIκBα、p-NF-κB和p-Akt相对表达量高于顺铂组和茶黄素组(P均<0.05)。结论茶黄素能够增强宫颈癌细胞对顺铂的敏感性,其机制可能与增加凋亡相关蛋白Cleaved Caspase 3、Cleaved Caspase 9、PARP表达及抑制NF-κB信号通路相关蛋白p-IκBα、p-NF-κB和p-Akt表达有关。
Objective To investigate the effects of theaflavins on the sensitivity of human cervical cancer HeLa cells to cisplatin and to explore the possible mechanism. Methods（1） HeLa cells in the logarithmic phase were inoculated on96-well plates,and then were divided into the observation groups 1-9 and the control groups 1-9,with three holes. The control groups 1-9 were treated with 0,5,10,15,20,25,30,35,40 and 50 μg/mL cisplatin,respectively,for 24 h.The observation groups 1-9 were treated with 10 μg/mL theaflavins,and then were treated with 0,5,10,15,20,25,30,35,40 and 50 μg/mL cisplatin,respectively,for 24 h. MTT assay was applied to detect the OD490,the half maximal inhibitory concentration（ IC（50）） was calculated in the observation group and the control groups.（2） HeLa cells in the logarithmic phase were inoculated on 96-well plates,and then were divided into the control group,the cisplatin group,the theaflavins group,and the combination group,with three holes in each. The control group was not treated with drug,the cisplatin group was treated with 10 μg/mL cisplatin,the theaflavins group was treated with 10 μg/mL theaflavins,and the combination group was treated with 10 μg/mL cisplatin and 10 μg/mL theaflavins,for 24 h. The apoptosis-related proteins Cleaved Caspase-3,Cleaved Caspase-9,PARP,and the NF-κB signal pathway-related proteins p-IκBα,p-NF-κB,and p-Akt were detected by Western blotting. Results（1） The cell survival rates in the control groups 1-9 were 102. 54% ± 4. 21%,98. 32% ± 2. 46%,96. 11% ± 3. 64%,90. 36% ± 3. 91%,86. 29% ± 3. 17%,81. 46% ± 3. 61%,78. 92% ± 4. 18%,70. 63% ± 4. 25%,and 63. 28% ± 4. 21%,respectively,and the IC50 value of HeLa cells to cisplatin was 25 μg/mL; the cell survival rates in the observation groups 1-9 were 102. 54% ± 4. 21%,80. 21% ± 3. 82%,78. 92% ± 4. 24%,77. 93% ± 3. 95%,68. 26% ± 3. 24%,50. 36% ± 4. 29%,45. 32% ± 4. 74%,40. 25% ± 3. 21%,and 35. 37% ±3. 16%,respectively,and the IC50 was 10 μg/mL.（2） The relative expression levels of Cleaved Caspase-3,Cleaved Caspase-9,and PARP in the cisplatin group,the theaflavins group,and the combination group were much higher than those in the control group（ all P〈0. 05）. The relative expression levels of Cleaved Caspase-3,Cleaved Caspase-9,and PARP were much higher than those in the cisplatin group and the theaflavins group（ all P〈0. 05）. No significant difference was found in the relative expression of p-IκBα,p-NF-κB,and p-Akt between the cisplatin group and the control group（ all P〈0. 05）. The relative expression levels of p-IκBα,p-NF-κB,and p-Akt in the combination group were higher than those in the cisplatin group and the theaflavins group（ all P〈0. 05）. Conclusion Theaflavins can enhance the sensitivity of cervical cancer cells to cisplatin by up-regulating the apoptosis-related proteins Cleaved Caspase-3,Cleaved Caspase-9,and PARP,and inhibiting the NF-κB signal pathway-related proteins p-IκBα,p-NF-κB,and p-Akt.