布比卡因通过调控miR-381-3p/HERC4表达抑制肺癌细胞增殖、迁移和侵袭  

Bupivacaine inhibits proliferation,migration and invasion of lung cancer cells by regulating miR-381-3p/HERC4 expression

作　　者：鹿素青[1] 秦志祥[1] 牛香兰[2] Lu Suqing;Qin Zhixiang;Niu Xianglan(Department of Anesthesiology,Heping Hospital Affiliated to Changzhi Medical College,Changzhi,Shanxi 046000,China;Department of Histology and Embryology,Changzhi Medical College,Changzhi,Shanxi 046000,China)

机构地区：[1]长治医学院附属和平医院麻醉科,山西长治046000 [2]长治医学院组织胚胎学教研室,山西长治046000

出　　处：《中华生物医学工程杂志》2020年第1期31-36,共6页Chinese Journal of Biomedical Engineering

摘　　要：目的探讨布比卡因对肺癌细胞增殖、迁移和侵袭的影响及其机制。方法分别使用浓度为100(BUP组)、300、600、1000μmol/L的布比卡因处理H1299细胞,未使用布比卡因处理的设为对照组。分别将miR-NC、miR-381-3p、si-NC、si-HERC4、anti-miR-NC、anti-miR-381-3p转染至H1299细胞中,其中转染anti-miR-NC和anti-miR-381-3p的细胞再使用100μmol/L布比卡因进行处理,分别记为miR-NC组、miR-381-3p组、si-NC组、si-HERC4组、BUP+anti-miR-NC组、BUP+anti-miR-381-3p组。qRT-PCR检测miR-381-3p和HERC4 mRNA的表达水平,免疫印迹法检测细胞周期蛋白D1(CyclinD1)、基质金属蛋白酶2(MMP-2)、MMP-9、p21等蛋白表达,MTT法检测细胞增殖抑制率,Transwell实验检测细胞迁移和侵袭,双荧光素酶报告基因检测实验检测miR-381-3p对HERC4的靶向调控作用。结果与对照组比较,随着布比卡因浓度增加,细胞增殖抑制率显著升高,细胞迁移和侵袭数量显著降低;且miR-381-3p表达水平显著升高,HERC4 mRNA和蛋白表达水平显著降低(均P<0.05)。双荧光素酶报告基因检测实验和免疫印迹显示,miR-381-3p靶向负调控HERC4的表达。过表达miR-381-3p或抑制HERC4表达均可抑制肺癌细胞的增殖、迁移和侵袭,而下调miR-381-3p表达可逆转布比卡因对肺癌细胞增殖、迁移和侵袭的抑制作用。结论布比卡因可抑制肺癌细胞的增殖、迁移和侵袭,其机制可能与调控miR-381-3p/HERC4的表达有关。Objective To investigate the effect of bupivacaine on the proliferation,migration and invasion of lung cancer cells and its mechanism.Methods H1299 cells were treated with bupivacaine at doses of 100(BUP group),300,600 and 1000μmol/L.Untreated H1299 cells were assigned to the control group.miR-NC,miR-381-3p,si-NC,si-HERC4,anti-miR-NC,and anti-miR-381-3p were respectively transfected into H1299 cells.Specifically,the cells transfected with anti-miR-NC and anti-miR-381-3p were further treated with 100μmol/L bupivacaine.Thereby,miR-NC,miR-381-3p,si-NC,si-HERC4,BUP+anti-miR-NC,and BUP+anti-miR-381-3p groups were set.The expression of miR-381-3p and HERC4 mRNA were detected by qRT-PCR.Western blotting was used to detect the protein expression of Cyclin D1,matrix metalloproteinase 2(MMP-2),MMP-9 and p21;MTT assay was used to measure the inhibition rate of cell proliferation;Transwell assay was used to detect the cell invasion and migration;Dual luciferase reporter gene assay was used to examine the targeted regulation of HERC4 by miR-381-3p.Results Compared with the control group,the inhibition rate of lung cancer cells increased significantly,and the number of migrating and invasive cells decreased significantly,the expression level of miR-381-3p significantly increased,and the expression levels of HERC4 mRNA and protein decreased,along with higher bupivacaine concentration(all P<0.05).Dual luciferase reporter assay and Western blotting showed that miR-381-3p negatively target-regulated HERC4 expression.Overexpression of miR-381-3p or suppression of HERC4 expression may inhibit proliferation,migration and invasion of lung cancer cells.Down-regulated miR-381-3p expression reversed the inhibitory effect of bupivacaine on proliferation,migration and invasion of lung cancer cells.Conclusion Bupivacaine may inhibit the proliferation,migration and invasion of lung cancer cells,and its mechanism may be related to the regulation of miR-381-3p/HERC4 expression.

关 键 词：布比卡因 微RNA-381-3p HERC4 肺癌 增殖 迁移 侵袭 

分 类 号：R734[医药卫生—肿瘤] R614[医药卫生—临床医学]