利多卡因通过LncRNA H19/miR-671-5p分子轴调控胃癌细胞增殖、迁移及侵袭

Lidocaine regulates proliferation,migration and invasion of gastric cancer cells through molecular axis of LncRNA H19/miR-671-5p

目的:探讨利多卡因对胃癌细胞增殖、迁移及侵袭的影响及其对LncRNA H19/miR-671-5p的调控机制。方法:体外培养胃癌细胞AGS,使用不同浓度的利多卡因处理细胞;MTT检测细胞增殖;Transwell小室实验检测细胞迁移及侵袭;qRT-PCR检测H19及miR-671-5p的表达量;双荧光素酶报告实验验证H19与miR-671-5p的靶向关系;将pcDNA-H19及其阴性对照pcDNA转染至AGS细胞,使用利多卡因处理细胞,采用上述方法检测细胞增殖、迁移及侵袭;Western blot检测增殖标记蛋白细胞增殖核抗原-67(Ki-67)、增殖细胞核抗原(PCNA)、上皮型钙黏蛋白(E-cadherin)、神经型钙黏蛋白(N-cadherin)的表达量。结果:与Con组相比,利多卡因不同剂量可明显降低细胞活力和Ki-67、PCNA、N-cadherin蛋白水平及H19的表达量(P<0.05),并可减少迁移及侵袭细胞数(P<0.05),提高E-cadherin蛋白水平及miR-671-5p的表达量(P<0.05);双荧光素酶报告实验证实H19可靶向结合miR-671-5p;H19过表达可明显逆转利多卡因对细胞增殖、迁移及侵袭的作用。结论:利多卡因可能通过抑制H19表达及促进miR-671-5p表达从而抑制胃癌细胞的增殖、迁移及侵袭。

Objective:To explore the effect of lidocaine on proliferation,migration and invasion of gastric cancer cells and its regulatory mechanism on LncRNA H19/miR-671-5p.Methods:Gastric cancer cells AGS were cultured in vitro,and cells were treated with different concentrations of lidocaine.MTT was used to detect cell proliferation.Transwell cell test was used to detect cell migration and invasion.qRT-PCR was used to detect expression levels of H19 and miR-671-5p.Dual luciferase report experiment was used to verified targeting relationship of H19 and miR-671-5p.pcDNA-H19 and its negative control pcDNA were transfected into AGS cells,the cells were treated with lidocaine,and cell proliferation,migration and invasion were detected by the above method.Western blot was used to detect expressions of Ki-67,PCNA,E-cadherin and N-cadherin.Results:Compared with Con group,different doses of lidocaine could significantly reduce cell viability,protein levels of Ki-67,PCNA,N-cadherin and expression level of H19(P<0.05),and reduce the number of migration and invasion cells(P<0.05),increase protein level of E-cadherin and expression of miR-671-5p(P<0.05).Dual luciferase report experiment confirmed that H19 could target miR-671-5p.H19 overexpression could obviously reverse the effects of lidocaine on cell proliferation,migration and invasion.Conclusion:Lidocaine may inhibit proliferation,migration and invasion of gastric cancer cells by inhibiting expression of H19 and promoting expression of miR-671-5p.