Rab26对小细胞肺癌H446细胞增殖和迁移的影响

Effects of Rab26 on the proliferation and migration of small cell lung cancer cells H446

目的探讨Rab26对小细胞肺癌H446细胞增殖和迁移的影响。方法设立Rab26质粒过表达组及Rab26 siRNA组,培养H446细胞,分别将含有Rab26过表达质粒和Rab26 siRNA瞬时转染H446细胞,同时设立空白对照组。48 h后,流式细胞仪检测转染效率,同时检测每组细胞中Rab26在mRNA及蛋白水平表达情况; MTT检测每组细胞的增殖情况,划痕试验观察每组细胞迁移的速度。结果Rab26过表达质粒组转染效率为(76.8±4.3)%,Rab26 siRNA组转染效率为(79.5±3.57)%,均为有效转染;转染48 h后,发现Rab26质粒过表达组中Rab26在mRNA和蛋白水平均较空白对照组表达水平显著上调(P<0.05),siRNA组结果显示Rab26 mRNA及蛋白表达水平显著下调(P<0.05); MTT检测结果显示转染24 h和48 h后,空白对照组细胞活力分别为(56.4±3.23)%、(100±4.31)%; Rab26质粒过表达组细胞活力分别为(42.5±3.59)%、(62. 3±2. 97)%; Rab26 siRNA组细胞活力分别为(75±3. 86)%、(123.4±5.66)%,即Rab26质粒过表达组细胞活力较空白组显著降低(P<0.05),而Rab26 siRNA组细胞活力较空白组显著升高(P<0.05),Rab26可抑制H446细胞的增殖;划痕实验结果显示转染24 h和48 h后,空白对照组细胞迁移率分别为(0.53±0.03)μm/min、(0.32±0.04)μm/min; Rab26质粒过表达组细胞迁移率分别为(0.21±0.04)μm/min、(0.22±0.04)μm/min; Rab26 siRNA组细胞迁移率分别为(0.61±0.02)μm/min、(0.42±0.03)μm/min,即Rab26质粒过表达组细胞迁移率较空白组显著较少(P<0.05),而Rab26 siRNA组细胞迁移率较空白组显著增加(P<0.05),Rab26可抑制H446细胞的迁移。结论 Rab26可抑制小细胞肺癌H446细胞的增殖和迁移,故调控Rab26的表达有望成为小细胞肺癌治疗的新靶点。

Objective To explore the effect of Rab26 on the Proliferation and migration of human in small cell lung cancer cells(H446).Methods The H446 cells were transfected with over-expression vector of Rab26,or Rab26 siRNA.Blank group was used as a negative control,respectively.After transfection 48 h,the protein level and mRNA of Rab26 were determined by RT-PCR and Western blot.Proliferation of the H446 cells was assayed by flow cytometry and the migration rate was detected by the scratch test.Results The transfection efficiency of Rab26 overexpression plasmid group was(76.8±4.3)%and Rab26 siRNA group was(79.5±3.57)%.After transfection 48 hours,the expression of Rab26 in overexpression group was significantly higher than control group in mRNA and protein level(P<0.05),however,the Rab siRNA group showed that the expression of Rab26 was significantly lower than control group(P<0.05).MTT assay showed that the cell viability of blank control group was(56.4±3.23)%,(100±4.31)%,Rab26 overexpression group was(42.5±3.59)%,(62.3±2.97)%and Rab26 siRNA group was(75±3.86)%,(123.31)%in 24 and 48 hours,respectively.The cell viability of Rab26 overexpression group was significantly lower than that of blank group(P<0.05),while Rab26 siRNA group was significantly higher than that of blank group(P<0.05).Scratch test showed that the cell migration rate of blank control group was(0.53±0.03)μm/min and(0.32±0.04)μm/min,Rab26 plasmid overexpression group was(0.21±0.04)μm/min and(0.22±0.04)μm/min,the Rab26 siRNA group was(0.61±0.02)μm/min,(0.42±0.03)um/min after 24 and 48 hours.The cell migration rates of Rab26 plasmid overexpression group was significantly lower than the blank group,but Rab26 siRNA group was significantly higher than the blank group(P<0.05).Conclusions Rab26 is associated with the cellular proliferation and migration in H446 cell.Rab26 may be a novel therapy target for the treatments of small cell lung cancer.