Expression Of Vasohibin-1 In Prostate Cancer And Preliminary Study On Its Biological Effects

Objectives: To discover the expression of vasohibin-1(VASH-1)in prostate cancer tissues,and explore the correlation between VASH-1,CD34,vascular endothelial growth factor-A(VEGF-A)and hypoxia inducible factor-1?(HIF-1?).Preliminary find the effect of VASH-1 on angiogenesis.Then study the expression of VASH-1,VEGF-A and HIF-1? in prostate cancer cell lines LNCa P,C4-2,PC3 and DU145,and to investigate the effect of VASH-1 overexpression on the proliferation and invasion of prostate cancer cells.Finally confirm that whether VASH-1 can be used as a new indicator to evaluate and predict the progression of prostate cancer,and whether it has anti-tumor effects.Methods:(1)60 cases of prostate cancer were stained by immunohistochemical staining,calculate the average optical density(AOD)of VASH-1,VEGF-A,HIF-1? by Image Plus 6.0.And the staining of CD34 was evaluated by microvessel density(MVD).The Spearman rank test was used to analyze the correlation between those factors.(2)Prostate cancer cell lines LNCa P,C4-2,PC3 and DU145 were cultured in vitro.Extract the cellular protein and RNA.The expression of VASH-1,VEGF-A and HIF-1? in the four cell lines were detected by Western Blot,PCR and Real-time PCR respectively.Two prostate cancer cell lines with low expression level of VASH-1 were then transfected with the recombinant plasmid p Receiver-M61-VASH-1.The effect of overexpression of VASH-1 on the proliferation and invasion of prostate cancer cells was investigated by MTT assay and Transwell invasion assay.Results:(1)VASH-1 mainly expressed in the cytoplasm of vascular endothelial cells and the prostate cancer cells,and its expression in the cytoplasm of prostate cancer cells was significantly higher than that in the vascular endothelial cell cytoplasm;CD34 is expressed in the cytoplasm of vascular endothelial cells;VEGF-A is mainly expressed in the cytoplasm and nucleus of vascular endothelial cells and cancer cells;HIF-1? is mainly expressed in the nucleus of cancer cells.The expression of the four factors were both significant correlated with the Gleason Score,p T stage and the distant metastasis(P<0.05),but there was no significant correlation with age and PSA level(P>0.05).The expression of VASH-1,CD34,VEGF-A and HIF-1? had asignificant positive correlation(P<0.05).(2)The expression of VASH-1,VEGF-A and HIF-1? in PC3 and DU145 cell lines were higher than that in LNCa P and C4-2 cell lines by Western Blot,PCR and Real-time PCR.After transfection of LNCa P and C4-2 with p Receiver-M61-VASH-1,the expression of VASH-1 in overexpression group was significantly higher than that in normal cell group and control group by Western Blot(P<0.05).Then,MTT assay showed that the optical density(OD)of overexpression group was significantly lower than that of normal cell group and control group at 48 hours and 72 hours(P<0.05);Transwell invasion assay showed that the number of cells transfected with VASH-1 was higher than that of normal cell group and control group,and the difference were statistically significant(P<0.05).Conclusion:(1)In prostate cancer,VASH-1 expression not only in vascular endothelial cells,but also in a large number of cancer cells,and with the VASH-1 expression levels increased,suggesting increased degree of malignancy of prostate cancer.The expression of VASH-1 was highly consistent with the expression of CD34,VEGF-A and HIF-1? in prostate cancer tissues.It was proved that VASH-1 had a certain effect on angiogenesis.(2)The recombinant plasmid p Receiver-M61-VASH-1 can promote the expression of VASH-1 in LNCa P and C4-2 cell lines.(3)Overexpression of VASH-1 can effectively inhibit the proliferation and invasion of LNCa P and C4-2.(4)In summary,VASH-1 may play an important role in the progression of prostate cancer angiogenesis.It is expected to be a new indicator to evaluate and predict the progression of prostate cancer,and has a certain anti-tumor effect.