Bojana B. Ilic, Jadranka A. Antic, Jovana Z. Bankovic, Ivana T. Milicevic, Gordana S. Rodic, Dusan S. Ilic, Cane D. Tulic, Vera N. Todorovic and Svetozar S. Damjanovic
Background: Sporadic clear-cell renal cell carcinoma (ccRCC) is associated with mutations in the VHL gene, upregulated mammalian target of rapamycin (mTOR) activity and glycolytic metabolism. Here, we analyze the effect of VHL mutational status on the expression level of mTOR, eIF4E-BP1, AMPK, REDD1, and PDK3 proteins. Methods: Total proteins were isolated from 21 tumorous samples with biallelic inactivation, 10 with monoallelic inactivation and 6 tumors with a wild-type VHL (wtVHL) gene obtained from patients who underwent total nephrectomy. The expressions of target proteins were assessed using Western blot. results: Expressions of mTOR, eIF4EBP1 and AMPK were VHL independent. Tumors with monoallelic inactivation of VHL underexpressed REDD1 in comparison to wtVHL tumors (P = 0.042), tumors with biallelic VHL inactivation (P < 0.005) and control tissue (P = 0.004). Additionally, REDD1 expression was higher in tumors with VHL biallelic inactivation than in control tissue (P = 0.008). Only in wt tumor samples PDK3 was overexpressed in comparison to tumors with biallelic inactivation of VHL gene (P = 0.012) and controls (P = 0.016). In wtVHL ccRCC, multivariate linear regression analysis revealed that 97.4% of variability in PDK3 expression can be explained by variations in AMPK amount. Conclusion: Expressions of mTOR, eIF4EBP1 and AMPK were VHL independent. We have shown for the first time VHL dependent expression of PDK3 and we provide additional evidence that VHL mutational status affects REDD1 expression in sporadic ccRCC.
Yong Han, Bingyi Shi, Ming Cai, Xiaoguang Xu, Liang Xu, Qiang Wang, Wenqiang Zhou and Li Xiao
CD20 Expression in the Transplanted Kidney of Patients with Graft Loss and Transient Allograft Dysfunction
This study aimed to explore the relationship between the infiltration of CD20+ B cells and the survival time of a renal allograft and to investigate the role of infiltrated B cells in the rejection of the renal allograft. A total of 40 patients with renal allograft loss due to refractory rejection and 20 patients with transient renal allograft dysfunction were recruited. Renal biopsy was done and CD20 expression was detected by immunohistochemistry. In addition, the survival time of the renal allograft was also obtained. The relationships between the CD20 expression and the survival time of the renal allograft and graft loss due to rejection were analyzed. The associations of gender, age and clinicopathogical types with the CD20 expression were also investigated. The proportion of patients positive for CD20 in the transplanted kidney was higher in patients receiving nephrectomy of the allograft due to rejection than in those with transient allograft dysfunction. The diffuse infiltration of CD20+ B cells was considered as positive staining. In 40 samples from patients with graft loss, 19 had diffuse infiltration of CD20+ B cells (47.5%). In 19 patients positive for CD20, hyperacute rejection was found in 1 patient, acute rejection in 5 and chronic rejection in 13. Statistical analysis showed the CD20 expression was not associated with the age and gender of donors and recipients, regimen for immunosuppressive treatment, cold/warm ischemia time and secondary transplantation. CD20+ B cell infiltration predicts a poor prognosis of patients with kidney transplantation and is one of the risk factors of graft loss.
). Indeed, in patients with chronic renal failure, ADMA is considered as a future marker of mortality and cardiovascular disease ( 49 ). SDMA was confirmed as an endogenous marker of renal failure that correlated directly with parameters of renal failure ( 53 , 54 ). After total nephrectomy in a rat model, the circulating level of SDMA increased rapidly in parallel with blood urea nitrogen and creatinine ( 55 ). SDMA can indirectly affect NO synthesis by two mechanisms that depend on interfering with the intracellular uptake of arginine: either by inhibiting renal