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利多卡因调控miR 15a 5p对肝癌细胞生物学功能的影响及其机制
赵铤,赵全丰,丁汉琳
0
(湖北江汉油田总医院麻醉科;襄阳市中心医院·湖北文理学院附属医院麻醉科)
摘要:
【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〖JP2〗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〖JP〗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。
关键词:  利多卡因;miR 15a 5p;肝癌;肝癌细胞HepG2  凋亡;迁移;侵袭
DOI:
基金项目:湖北省卫计委基金项目(WJ2015MB184)
Effect of lidocaine on miR 15a 5p on apoptosis, migration and invasion of liver cancer cells and its mechanism
ZHAO Ting,ZHAO Quanfeng,DING Hanlin
(Department of Anesthesiology, General Hospital of Jianghan Oilfield;Department of Anesthesiology, Xiangyang Central Hospital, The Affiliated Hospital of Hubei Academy of Arts and Sciences)
Abstract:
【Abstract】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.
Key words:  Lidocaine  miR 15a 5p  Liver cancer  HepG2  Apoptosis  Migrate  Invasion

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