1.Quantitative detection of DD3 mRNA and PSA mRNA expressions in prostate cancer tissues by realtime fluorescent quantitative reverse transcription polymerase chain reaction
Xiaolu MAO ; Zhihua TAO ; Wei XU ; Xiaodong CHEN ; Zhanguo CHEN ; Zhiliang WENG ; Yuanping HV ; Xiuling WU ; Xiaohua ZHANG ; Hui XLE ; Ouchen WANG ; Qitong SONG ; Chengdi LI ; Kaiyuan YU
Chinese Journal of Urology 2001;0(09):-
Objective To study the expressions of DD3 mRNA and PSA mRNA in the prostate tissues and its diagnostic value in prostate cancer (PCa). Methods DD3 mRNA and PSA mRNA were detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction ( FQ-RT-PCR) based on Taqman technique in the tissues of 21 cases of PCa and 39 cases of BPH. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic efficacy of DD3 mRNA, PSA mRNA and DD3 mRNA/PSA mRNA. Results The expressions of DD3 mRNA and PSA mRNA, and DD3 mRNA/ PSA mRNA were significantly higher in PCa tissues than those in BPH tissues ( P 0.05 for all). The AUC-ROC of DD3 mRNA,PSA mRNA and DD3 mRNA/PSA mRNA were 0. 937 (95% CI,0. 879 -0. 995) , 0.755(95% CI,0.629 -0.880) and 0.839 (95%CI,0.738 -0.940),respectively. The sensitivity for DD3 mRNA,PSA mRNA and DD3 mRNA/PSA mRNA was 90. 5% ,81. 0% and 81. 0% , respectively, and the specificity was 85.0% ,62.0% and 66.7% at cutoff value of 1.4?105 copies/mg tissue,3.0?107 copies/ mg tissue and 5. 0?10-3,respectively. The sensitivity and specificity of simultaneous detection for DD3 mRNA and PSA mRNA were 100% and 85.0%. Conclusions Both DD3 mRNA and PSA mRNA expressions were significantly higher in PCa tissues than those in BPH tissues; and the quantitative detection of DD3 mRNA is more helpful for the diagnosis. The simultaneous detection of DD3 mRNA and PSA mRNA can improve the sensitivity in the diagnosis of PCa.
2.Fibroblast Growth Factor 21 Attenuates Diabetes-Induced Renal Fibrosis by Negatively Regulating TGF-β-p53-Smad2/3-Mediated Epithelial-to-Mesenchymal Transition via Activation of AKT
Sundong LIN ; Lechu YU ; Yongqing NI ; Lulu HE ; Xiaolu WENG ; Xuemian LU ; Chi ZHANG
Diabetes & Metabolism Journal 2020;44(1):158-172
BACKGROUND:
Epithelial-to-mesenchymal transition (EMT) is required for renal fibrosis, which is a characteristic of diabetic nephropathy (DN). Our previous study demonstrated that fibroblast growth factor 21 (FGF21) prevented DN associated with the suppressing renal connective tissue growth factor expression, a key marker of renal fibrosis. Therefore, the effects of FGF21 on renal fibrosis in a DN mouse model and the underlying mechanisms were investigated in this study.
METHODS:
Type 1 diabetes mellitus was induced in C57BL/6J mice by intraperitoneal injections of multiple low doses of streptozotocin. Then, diabetic and non-diabetic mice were treated with or without FGF21 in the presence of pifithrin-α (p53 inhibitor) or 10-[4′-(N,N-Diethylamino)butyl]-2-chlorophenoxazine hydrochloride (10-DEBC) hydrochloride (Akt inhibitor) for 4 months.
RESULTS:
DN was diagnosed by renal dysfunction, hypertrophy, tubulointerstitial lesions, and glomerulosclerosis associated with severe fibrosis, all of which were prevented by FGF21. FGF21 also suppressed the diabetes-induced renal EMT in DN mice by negatively regulating transforming growth factor beta (TGF-β)-induced nuclear translocation of Smad2/3, which is required for the transcription of multiple fibrotic genes. The mechanistic studies showed that FGF21 attenuated nuclear translocation of Smad2/3 by inhibiting renal activity of its conjugated protein p53, which carries Smad2/3 into the nucleus. Moreover pifithrin-α inhibited the FGF21-induced preventive effects on the renal EMT and subsequent renal fibrosis in DN mice. In addition, 10-DEBC also blocked FGF21-induced inhibition of renal p53 activity by phosphorylation of mouse double minute-2 homolog (MDM2).
CONCLUSION
FGF21 prevents renal fibrosis via negative regulation of the TGF-β/Smad2/3-mediated EMT process by activation of the Akt/MDM2/p53 signaling pathway.