摘要
目的探讨猕猴桃根提取物抗肝细胞癌(HCC)的作用及相关机制。方法选取HCC细胞系HepG2和HUH7,根据猕猴桃根浓度分为0μg/ml(对照组)、50μg/ml、100μg/ml、200μg/ml组。采用噻唑蓝(MTT)和克隆实验检测细胞增殖;流式细胞术检测细胞周期和细胞凋亡;划痕实验和Transwell实验研究细胞的侵袭和转移;Real-time PCR和Western blot检测相关基因和蛋白表达。结果猕猴桃根对HCC增殖的抑制作用存在剂量和时间依赖性。克隆实验发现,与对照组比较,不同浓度猕猴桃根抑制HepG2和HUH7细胞增殖,且随浓度的增加,抑制作用逐渐增强,差异有统计学意义(P<0.05)。流式细胞术检测结果发现,与对照组比较,50μg/ml、100μg/ml猕猴桃根促进细胞由G1期进入S期,抑制DNA复制,促进HepG2和HUH7细胞凋亡,差异均有统计学意义(P<0.05)。划痕实验结果发现,与对照组比较,猕猴桃根显著促进HepG2和HUH7细胞迁移,具有剂量依懒性(P<0.05)。猕猴桃根促进Akt基因和蛋白表达,抑制p-Akt表达,抑制Akt磷酸化比例,促进PTEN基因和蛋白表达,差异有统计学意义(P<0.05)。结论猕猴桃根通过p-Akt/PTEN调控HCC细胞增殖、周期停滞及细胞凋亡。
Objective To investigate the hepatocellular carcinoma(HCC)effect of kiwifruit root extract and its related mechanism.Methods The HCC cell lines HepG2 and Huh7 were selected and divided into 0 μg/ml(control group),50 μg/ml,100 μg/ml and 200 μg/ml groups according to kiwifruit root concentration.MTT and clone experiments were used to detect cell proliferation,flow cytometry was used to detect cell cycle and apoptosis,scratch experiment and Transwell experiment were used to study cell invasion and metastasis,Real-time PCR and Western blot were used to detect the expression of related genes and proteins.Results The inhibition of kiwifruit root on HCC proliferation was dose-dependent and time-dependent.The cloning experiment found that kiwifruit root inhibited the proliferation of HepG2 and HUH7 cells at different concentrations compared with the control group,and the inhibitory effect gradually increased with the increase of concentration,the difference was statistically significant(P<0.05).The results of flow cytometry showed that,compared with the control group,50 μg/ml,100 μg/ml kiwifruit root promoted cells from G1 phase to S phase,inhibited DNA replication,and promoted apoptosis of HepG2 and HUH7 cells(P<0.05).The results of the scratch experiment showed that,compared with the control group,kiwifruit root significantly promoted the migration of HepG2 and HUH7 cells,with dose-dependent properties(P<0.05).Kiwifruit root promoted the expression of Akt gene and protein,inhibited the expression of p-Akt,inhibited the phosphorylation ratio of Akt,and promoted the expression of PTEN gene and protein(P<0.05).Conclusion Kiwifruit root regulates the proliferation,cycle stagnation and apoptosis of HCC cells through p-Akt/PTEN.
作者
徐莹
任雪
佟昌慈
武忠宝
柳云恩
阎英
XU Ying;REN Xue;TONG Chang-ci;WU Zhong-bao;LIU Yun-en;YAN Ying(Department of Radiotherapy,General Hospital of Northern Theater Command,Shenyang 110016,China;Laboratory of Rescue Center of Severe Wound and Trauma PLA,Severe Trauma and Organ Protection Laboratory of Liaoning Province,Shenyang 110016,China)
出处
《临床军医杂志》
CAS
2020年第2期128-132,共5页
Clinical Journal of Medical Officers
基金
辽宁省科学技术计划项目(20170540981)。
作者简介
第一作者:徐莹(1982-),女,辽宁沈阳人,副主任医师,博士;通信作者:阎英,E-mail:yanyingdoctor@sina.com;通信作者:柳云恩,E-mail:148964918qq.com。
