氯普鲁卡因通过调控circRNA-ZKSCAN1表达抑制肝癌Huh-7细胞体外生长和转移的研究  

Chloroprocaine inhibits the growth and metastasis of liver cancer Huh-7 cells in vitro by regulating the expression of circRNA-ZKSCAN1

作　　者：曾聿理 雷发容 肖慧 邱德亮 谢静 吴寻[2] Zeng Yuli;Lei Farong;Hui Xiao;Qiu Deliang;Xie Jing;Wu Xun(Department of Anesthesiology,the First People's Hospital of Longquanyi District,Chengdu 610100,China;Department of General Surgery,the First People's Hospital of Longquanyi District,Chengdu 610100,China)

机构地区：[1]成都市龙泉驿区第一人民医院麻醉科,610100 [2]成都市龙泉驿区第一人民医院普外科,610100

出　　处：《中华细胞与干细胞杂志（电子版）》2024年第4期220-228,共9页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：四川省卫生和计划生育委员会科研课题(16PJ091)。

摘　　要：目的探讨氯普鲁卡因(CP)对人肝癌细胞Huh-7生物学行为的影响及其对circRNA-ZKSCAN1的调控作用。方法同剂量CP处理Huh-7细胞,后续实验处理:pcDNA、pcDNA-circRNA-ZKSCAN1分别转染至Huh-7细胞,si-NC、si-circRNA-ZKSCAN1分别转染至Huh-7细胞后用CP(10 mmol/L)处理;qRT-PCR法检测circRNA-ZKSCAN1表达量;CCK-8、平板克隆形成实验、划痕实验、Transwell小室分别检测细胞增殖、克隆形成、迁移及侵袭能力;Western blot检测各组细胞E-钙黏蛋白(E-cadherin)和N-钙黏蛋白(N-cadherin)蛋白的表达情况。两组间比较使用t检验,多组间比较使用单因素方差分析,进一步两两比较采用SNK-q检验。结果与对照组相比,CP低、中、高剂量干预后,细胞增殖抑制率[(24.20±2.21)%、(48.25±4.42)%、(65.19±5.49)%比(0.00±0.00)%]、E-cadherin蛋白水平、circRNA-ZKSCAN1表达量[1.51±0.13、2.11±0.22、2.92±0.27比1.00±0.00]升高(P<0.05),细胞克隆形成数、侵袭细胞数[(99.96±7.03)、(79.87±4.96)、(58.64±3.99)比(124.01±10.98)个]减少(P<0.05),划痕愈合率[(53.71±4.19)%、(40.61±4.58)%、(24.13±2.22)%比(67.15±5.17)%]、N-cadherin蛋白水平降低(P<0.05),且呈剂量依赖性;与转染pcDNA相比,转染pcDNA-circRNA-ZKSCAN1后细胞增殖抑制率[(57.19±5.27)%比(7.13±0.54)%]、E-cadherin蛋白水平升高(P<0.05),细胞克隆形成数、划痕愈合率[(31.11±3.01)%比(68.07±4.76)%]、侵袭数[(66.33±5.84)比(126.12±11.74)个]以及N-cadherin蛋白水平降低(P<0.05);转染si-circRNA-ZKSCAN1可减低CP对Huh-7细胞生物学行为的作用。结论CP可通过上调circRNA-ZKSCAN1表达而抑制肝癌Huh-7细胞生长、转移。Objective To investigate the effect of chloroprocaine(CP)on the biological behavior of human hepatoma cell Huh-7 and its regulation on circZKSCAN1.Methods Huh-7 cells were treated with different doses of CP,and subsequent experimental treatments:pcDNA and pcDNA-circZKSCAN1 were transfected into Huh-7 cells,respectively.si-NC and si-circZKSCAN1 were transfected into Huh-7 cells and treated with CP(10 mmol/L).QRT-PCR detected the expression of circZKSCAN1.CCK-8,plate clone formation assay,scratch assay,and Transwell chamber were used to detect cell proliferation,clone formation,migration and invasion abilities,respectively.Western blot was used to detect the expression of E-cadherin and N-cadherin.T test was used for comparison between the two groups,One-way analysis of variance was used for statistical comparison among multiple groups,and the SNK-q method was used for further pairwise statistical comparison.Results After the treatment of CP,the inhibition rate of cell proliferation[(24.20±2.21)%,(48.25±4.42)%,(65.19±5.49)%vs(0.00±0.00)%],the protein level of E-cadherin,and the expression of circZKSCAN1[1.51±0.13,2.11±0.22,2.92±0.27 vs 1.00±0.00]were increased(P<0.05),while the number of cell clones and invasive cells[(99.96±7.03),(79.87±4.96),(58.64±3.99)vs(124.01±10.98)pcs]was decreased(P<0.05),and the wound healing rate[(53.71±4.19)%,(40.61±4.58)%,(24.13±2.22)%vs(67.15±5.17)%],the protein level of N-cadherin were decreased(P<0.05),and they were dose-dependent.After transfection of pcDNA-circZKSCAN1,the inhibition rate of cell proliferation[(57.19±5.27)%vs(7.13±0.54)%]and the protein level of E-cadherin were higher than those of transfected pcDNA(P<0.05),while the number of clones and invasive cells[(31.11±3.01)%vs(68.07±4.76)%]was lower than those of transfected pcDNA(P<0.05),and the wound healing rate[(66.33±5.84)vs(126.12±11.74)pcs],the protein level of N-cadherin was lower than that of transfected pcDNA(P<0.05).Transfection of si-circZKSCAN1 could reduce the effect of CP on the biologica

关 键 词：氯普鲁卡因 circRNA-ZKSCAN1 肝癌 细胞增殖 迁移 侵袭 

分 类 号：R735.7[医药卫生—肿瘤]