ABSTRACT
Objective To investigate the roles and mechanisms of siRNA targeted lipocalin 2 (Lcn 2) gene silencing on hypoxia-induced cell apoptosis in retinal ganglion cells (RGC-5).Methods RGC-5 cells were subjected to hypoxic condition for various time duration (0 h,4 h,8 h,12 h,24 h,48 h),and quantitative real-time PCR and Western blot were used to evaluate the expression of Lcn 2 in mRNA and protein levels.Cells were divided into 4 groups:control group,hypoxia group,siNC + hypoxia group,in which cells were transfected with negative control for siRNA,and then subjected to hypoxic condition for 24 h,and siLcn 2 + hypoxia group,in which cells were transfected with Lcn 2 siRNA,and then subjected to hypoxic condition for 24 h).Next,cell apoptotic rate and Caspase-3 activity were detected using ELISA.The fluorescence intensity of reactive oxygen species (ROS) was assayed using DCFH-DA kit,and mitochondrial membrane potential assay kit was used to evaluate the mitochondrial membrane potential.Finally,the expression levels of cleaved-Caspase-3 (c-Caspase-3),cleaved-PARP (c-PARP),Bax,Bcl-2 and cytochrome C (Cyto C) were detected using Western blot.Results In the hypoxia group,Lcn 2 mRNA level at 4 h,8 h,12 h,24 h and 48 h was higher than that at 0 h (all P ＜0.05).Meanwhile,Lcn 2 protein level at 12 h,24 h and 48 h was also higher than that at 0 h (all P ＜ 0.05).Cell apoptotic rate in the hypoxia group (138.33％ ± 13.76％) was significantly higher than that in the control group (99.66％ ± 2.86％) (P ＜ 0.05),and siLcn 2 + hypoxia group (105.02％ ± 8.60％) was lower than siNC + hypoxia group (142.33％ ± 6.54％) (P ＜ 0.05).In addition,compared with the control group,the relative activity of Caspase-3 and the relative expression of c-Caspase-3 and c-PARP in the hypoxia group were all upregulated (all P ＜ 0.05);whereas the relative activity of Caspase-3 and the relative expression of c-Caspase-3 and c-PARP in the siLcn2 + hypoxia group were downregulated compared with the siNC + hypoxia group (all P ＜ 0.05).Moreover,the fluorescence intensity of ROS in the hypoxia group (4.26 0.63) was more enhanced than that in the control group (1.03 ± 0.11) (P ＜ 0.05),and the siLcn2 + hypoxia group (3.10 ± 0.24) was lower than siNC + hypoxia group (4.73 ± 0.26) (P ＜ 0.05).Furthermore,mitochondrial membrane potential,Cyto C expression and the relative ratio of Bax to Bcl-2 in the siLcn2 + hypoxia group was lower than those in the siNC + hypoxia group.Conclusion Lcn 2 Lcn2 gene silencing can inhibit cell apoptosis and ROS production induced by hypoxia,which may be achieved by suppressing mitochondrial apoptosis signaling pathway.
ABSTRACT
Objective To investigate the correlation between serum vascular endothelial growth factor(VEGF)and the fac-tors of depression in patients with primary liver cancer(HCC).Methods The retrospective analysis was about 43 cases of primary liver cancer patients of Shaanxi Provincial People's Hospital,from September 2015 to September 2016.The self-rat-ing depression scale was used to evaluate the patients with depression and non-depression group.By ELISA kits,test VEGF levels,analysis of depression,age,gender and serum VEGF concentration.Results The level of VEGF in patients with de-pression was 86.21±19.52 pg/ml,which was significantly higher than that in patients without depression(48.31±9.16 pg/ml)by t test(t=-2.968,P=0.005).The levels of serum VEGF in patients with different gender groups were meas-ured by t test(t=0.079,P=0.938).And the serum levels of VEGF were measured in patients with different age groups by multiple sets of variance analysis(F=0.618,P=0.655).For age and gender,there was no significant difference in serum VEGF level between HCC.Conclusion By HCC,it was related between depression and the level of serum VEGF,and it suggests that serum level of VEGF may become the prognosis of primary liver cancer detection index for these patients.Giv-ing psychological interventions earlier,may improve the prognosis of cancer patients with depression.