摘要
目的探讨利多卡因对肝癌细胞凋亡、迁移和侵袭的影响及其机制。方法使用不同浓度利多卡因作用于肝癌细胞HepG2,正常培养的HepG2细胞为对照组。流式细胞术检测细胞凋亡,蛋白质印迹法(Western blot)检测B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、E-钙黏蛋白(E-cadherin)、基质金属蛋白酶-2(MMP-2)蛋白表达,Transwell小室法检测细胞迁移和侵袭,qPCR检测细胞中miR-15a-5p表达。在细胞中转染miR-15a-5p、anti-miR-15a-5p及各自阴性对照的转染并使用0.1 mmol/L利多卡因处理细胞,观察其对HepG2细胞凋亡、迁移、侵袭的影响。结果与对照组相比,0.01、0.1和1 mmol/L利多卡因增加细胞凋亡率和Bax蛋白表达量(P<0.05),降低Bcl-2蛋白水平(P<0.05)。0.1 mmol/L利多卡因明显降低迁移细胞数、侵袭细胞数和MMP-2蛋白表达量(P<0.05),升高E-cadherin蛋白水平和miR-15a-5p表达量(P<0.05)。过表达miR-15a-5p增加细胞凋亡率、Bax和E-cadherin蛋白表达量(P<0.05),减少迁移细胞数、侵袭细胞数、Bcl-2和MMP-2蛋白表达量(P<0.05)。结论利多卡因通过调控miR-15a-5p表达抑制肝癌细胞迁移和侵袭,并诱导细胞凋亡。
Objective To explore the effects of lidocaine on apoptosis,migration and invasion of liver cancer cells and its mechanism.Methods Liver cancer cells HepG2 were treated with different concentrations of lidocaine.Cell apoptosis was determined by flow cytometry.The expressions of B cell lymphoma/leukemia-2(Bcl-2),Bcl-2 related X protein(Bax),E-cadherin and matrix metalloproteinase-2(MMP-2)were analyzed by Western blot.Transwell chambers method was used to measure cell migration,and invasion,and qPCR was applied to test the expression of miR-15a-5p in cells.The cells were transfected with miR-15a-5p,or anti-miR-15a-5p and treated with 0.1mmol/L lidocaine to investigate their effects on apoptosis,migration and invasion of HepG2 cells.Results Compared with the control group,0.01,0.1 and 1 mmol/L lidocaine greatly increased the apoptosis rate and the expression level of Bax protein(P<0.05),and significantly reduced the level of Bcl-2 protein(P<0.05).0.1 mmol/L lidocaine obviously decreased the number of migrating cells,the number of invading cells and the expression of MMP-2 protein(P<0.05),while evidently improved the expression of E-cadherin protein and miR-15a-5p(P<0.05).The overexpression of miR-15a-5p markedly raised the apoptosis rate,the expression levels of Bax and E-cadherin proteins(P<0.05),and dramatically declined migrating cells,invading cells,the expression levels of Bcl-2 and MMP-2 proteins(P<0.05).Inhibition of miR-15a-5p reversed lidocaine's roles in promoting apoptosis of HepG2 cells,Bax and E-cadherin protein expression,and reversed its roles in inhibiting cell migration,invasion,Bcl-2 and MMP-2 protein expression.Conclusion Lidocaine inhibits the migration and invasion of liver cancer cells and induces apoptosis by regulating the expression of miR-15a-5p.
作者
赵铤
赵全丰
丁汉琳
ZHAO Ting;ZHAO Quanfeng;DING Hanlin(Department of Anesthesiology, General Hospital of Jianghan Oilfield, Qianjiang 433124, Hubei, China;Department of Anesthesiology, Xiangyang Central Hospital, The Affiliated Hospital of Hubei Academy of Arts and Sciences, Xiangyang 441021, Hubei, China)
出处
《西部医学》
2020年第7期980-985,989,共7页
Medical Journal of West China
基金
湖北省卫计委基金项目(WJ2015MB184)。
