利多卡因通过SHH/GLI信号通路对胶质瘤细胞恶性生物学行为的影响

Effects of lidocaine on the malignant biological behavior of glioma cells through SHH/GLI signaling pathway

目的探讨利多卡因(Lid)调节SHH/GLI信号通路对胶质瘤细胞恶性生物学行为的影响。方法将U87细胞分为U87组、L-Lid组、M-Lid组、H-Lid组、PM组、H-Lid+PM组。U87组未做任何处理,L-Lid组、M-Lid组、H-Lid组分别用5、10、15 mmol/L Lid处理U87细胞,PM组用1μmol/L的SHH/GLI信号通路激活剂PM处理U87细胞,H-Lid+PM组用15 mmol/L Lid和1μmol/L的PM共同处理U87细胞。CCK8检测U87细胞增殖;流式细胞术检测U87细胞凋亡;Transwell检测U87细胞迁移、侵袭能力;Western blot检测SHH/GLI蛋白、凋亡蛋白表达;荷瘤小鼠模型验证Lid对U87细胞移植瘤的影响。结果与U87组相比,L-Lid组、M-Lid组、H-Lid组的OD值、U87细胞迁移、侵袭数量、Bcl-2、SHH、GLI3蛋白水平、小鼠肿瘤体积依次显著下降(P<0.05),凋亡率、Bax蛋白水平依次显著升高(P<0.05);而PM组OD值、U87细胞迁移、侵袭数量、Bcl-2、SHH、GLI3蛋白水平、小鼠肿瘤体积显著升高(P<0.05),凋亡率、Bax蛋白水平显著下降(P<0.05);PM消除了H-Lid对U87细胞恶性生物学行为的抑制作用。Lid处理后移植瘤小鼠存活数量增加,而PM处理后移植瘤小鼠存活数量减少,H-Lid+PM组与L-Lid组数量相同。结论Lid可能通过下调SHH/GLI信号通路抑制U87细胞增殖,促进细胞凋亡,进而抑制胶质瘤的进展。

Objective To investigate the impact of lidocaine(Lid)on malignant biological behavior of glioma cells by regulating SHH/GLI signal pathway.Methods The U87 cells were divided into U87 group,L-Lid group,M-Lid group,H-Lid group,PM group,and H-Lid+PM group.No treatment was performed on U87 cells in L-Lid group,M-Lid group,and H-Lid group.U87 cells were treated with 5,10,and 15 mmol/L Lid,respectively,while PM group was treated with 1 mmol/L LidμU87 cells were treated with mol/L of SHH/GLI signal pathway activator PM,while H-Lid+PM group was treated with 15 mmol/L Lid and 1μThe U87 cells were co treated with mol/L of PM.CCK8 was applied to detect U87 cell proliferation;flow cytometry was applied to detect U87 cell apoptosis;Transwell was applied to detect the migration and invasion abilities of U87 cells;Western blot was applied to detect the expression of SHH/GLI protein and apoptosis protein;the tumor-bearing mouse model was applied to verify the effect of Lid on U87 cell transplanted tumor.Results Compared with U87 group,the OD value,numbers of U87 cell migration and invasion,the protein levels of Bcl-2,SHH and GLI3,and tumor volume of mice in L-Lid group,M-Lid group and H-Lid group decreased obviously in turn(P<0.05),the apoptosis rate and Bax protein level increased obviously in turn(P<0.05);the OD value,numbers of U87 cell migration and invasion,the protein levels of Bcl-2,SHH and GLI3,and tumor volume of mice in PM group were obviously increased(P<0.05),the apoptosis rate and Bax protein level decreased obviously(P<0.05);PM eliminated the inhibitory effect of H-Lid on the malignant biological behavior of U87 cells.The survival number of transplanted tumor mice increased after Lid treatment,while the survival number of transplanted tumor mice decreased after PM treatment,and the number in H-Lid+PM group was the same as that in L-Lid group.Conclusion Lid may inhibit the proliferation of U87 cells,promote cell apoptosis,and then inhibit the progression of glioma by down-regulating SHH/GLI signal pathway.