6-姜烯酚对HPV阳性及阴性宫颈癌细胞侵袭及迁移的作用及其机制  

Effect and mechanism of 6-gingerol on invasion and migration of HPV-positive and negative cervical cancer cells

作　　者：赵行宇 张漠 朱志华 侯建成 刘勃 张巍 Zhao Xing-Yu;Zhang Mo;Zhu Zhi-Hua;Hou Jian-Cheng;Liu Bo;Zhang Wei(Basic College of Jilin Medical University,Jilin City,Jilin Province 132013,China)

机构地区：[1]吉林医药学院基础医学院,吉林省吉林市132013

出　　处：《解放军医学杂志》2020年第7期691-696,共6页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(21102055);吉林省科技厅科技发展项目(20190701062GH);吉林省教育厅“十三五”科学技术研究项目(JJKH20191062KJ)。

摘　　要：目的观察6-姜烯酚对人乳头瘤病毒(HPV)阳性及阴性人宫颈癌细胞侵袭及迁移的抑制作用,并探讨其作用机制。方法培养HPV阳性人宫颈癌细胞(HeLa)及HPV阴性人宫颈癌细胞(C33A),分别加入0、5、10、20、50μmol/L的6-姜烯酚,并以未处理组作为对照,继续培养24 h。确定6-姜烯酚的最佳浓度为10μmol/L。后续实验以10μmol/L的6-姜烯酚分别处理24、48及72 h,采用MTS法检测细胞增殖情况,以10μmol/L 6-姜烯酚用于细胞划痕实验检测6-姜烯酚对细胞迁移能力的影响,采用Transwell小室法检测6-姜烯酚对细胞侵袭能力的影响,Western blotting检测细胞中基质金属蛋白酶(MMP)-2、MMP-9、E-钙黏蛋白(E-cadherin)及N-钙黏蛋白(N-cadherin)的表达水平。结果MTS检测结果表明,HeLa及C33A细胞的活力均随6-姜烯酚浓度及作用时间的增加而逐渐降低,在各浓度及作用时间点,HeLa细胞的活力降低较C33A细胞更明显;Transwell侵袭实验结果显示,6-姜烯酚对HeLa细胞作用24 h及48 h,对C33A细胞作用48 h后,细胞的侵袭能力明显降低;划痕实验结果显示,6-姜烯酚对HeLa细胞作用24 h及48 h后,对C33A细胞作用48 h后,细胞的伤口愈合率明显降低。Western blotting检测结果显示,10μmol/L 6-姜烯酚处理HeLa细胞24 h后,细胞中E-cadherin的表达增加,而N-cadherin、MMP-2及MMP-9的表达下降,差异有统计学意义(P<0.05);10μmol/L 6-姜烯酚处理C33A细胞24 h后,细胞中E-cadherin的表达明显增加,而N-cadherin、MMP-2及MMP-9的表达与未处理组比较差异无统计学意义(P>0.05)。结论6-姜烯酚能抑制HPV阳性宫颈癌细胞的增殖并抑制细胞侵袭,其机制可能与6-姜烯酚影响上皮间充质转化相关蛋白的表达有关。Objective To observe the inhibitory effect of 6-gingerol on the invasion and migration of human papilloma virus(HPV)-positive and HPV-negative cervical cancer cells,and explore the possible mechanism.Methods Human HPV-positive cervical cancer cells(HeLa)and HPV-negative cells(C33A)were cultured,and added with 6-gingerol at different concentration of 0,5,10,20,50μmol/L,the untreated cells play as control,then cultured for 24 h.10μmol/L 6-gingerol was determined as the best concentration,then all the cells were treated with 10μmol/L 6-gingerol for 24 h,48 h and 72 h respectively.The cell proliferation was detected by MTS method,and cell scratch test was performed to detect the effect of 6-gingerol on cell migration ability.The effect of 6-gingerol on cellular invasion was detected by Transwell chamber.Western blotting was used to detect the expression of matrix metalloproteinase MMP-2,MMP-9,E-cadherin and N-cadherin.Results MTS assay showed that the activity of HeLa and C33A cells decreased with the increase of 6-gingerol concentration and action time,while the activity of HeLa cells decreased more significantly than that of C33A cells at different concentrations or time points.Transwell invasion chamber test showed that the HeLa cells treated with 6-gingerol for 24 h and 48 h,and C33A cells treated with 6-gingerol for 48 h,the cellular invasion ability decreased significantly.The scratch test revealed that the wound healing rate decreased significantly of HeLa cells 24 h and 48 h after 6-carbenol action and of C33A cells 48 h after 6-carbenol action.Western blotting results showed that,treating with 10μmol/L of 6-gingerol for 24 h,the expressions of E-cadherin increased and of N-cadherin,MM P-2 and MMP-9 declined in HeLa cells with statistical differences(P<0.05);while in C33A cells,the expressions of E-cadherin increased markedly,and of N-cadherin,MM P-2 and MMP-9 showed no significant differences(P>0.05)compared to that of unprocessed group.Conclusions 6-gingerol can inhibit the proliferation of HPV-positive

关 键 词：6-姜烯酚 宫颈癌 人乳头瘤病毒 肿瘤浸润 细胞运动 

分 类 号：R737.33[医药卫生—肿瘤]