1.Apoptosis of the prostate cancer cell line PC-3M induced by E2F decoy DNA
Tao WANG ; Anli JIANG ; Pengju ZHANG ; Tong CHEN ; Meilan HE ; Weiwen CHEN ; Jingti DENG ; Jianye ZHANG
Chinese Journal of Pathophysiology 2000;0(08):-
AIM: To observe the effect of E2F decoy DNA on proliferation and apoptosis of androgen-independent prostate cancer cell line PC-3M.METHODS: E2F decoy DNA,ARE decoy DNA and control decoy DNA were transfected into PC-3M cells with lipofectamine,respectively.Their effects on cell proliferation were detected by MTT assay.The changes of cell morphology were observed by inverted phase contrast microscope.The cell apoptotic rate was determined by flow cytometry(FCM) analysis and chromosome DNA ladder was detected by DNA gel electrophoresis.The expression of c-Myc mRNA and cyclin D1 mRNA was detected by RT-PCR.The protein levels of c-Myc and cyclin D1 were detected by Western blotting.RESULTS: The growth of PC-3M cells was inhibited after transfection.The transfected PC-3M cells displayed typical apoptotic morphological changes.The apoptotic rate was 26.35% and DNA ladder was observed after transfection.The expression of c-Myc and cyclin D1 were inhibited.CONCLUSION: These results indicate that E2F decoy DNA induces apoptosis of androgen-independent prostate cancer cell lines PC-3M and inhibits cell proliferation via inhibiting expression of c-Myc and cyclin D1.
2.Correlation of imbalance of urinary exosome Th1/Th2 with diabetic nephropathy
Aili SUN ; Yihong NI ; Guangju GUAN ; Haoping DENG ; Yuantao LIU ; Shihong CHEN ; Fudun SUN ; Xianghua ZHUANG ; Xiaoyan HU ; Jingti DENG
Chinese Journal of Nephrology 2011;27(9):637-640
Objective To examine the correlation of imbalance of urinary exosome Th1/Th2 with diabetic nephropathy (DN).Methods A total of 120 patients with type 2 diabetes mellitus (DM) and 30 healthy volunteers as control were enrolled in the study.According to urinary albumin/creatinine ratio (UACR),type 2 diabetes mellitus patients were divided into 3 groups:diabetes mellitus without nepbropathy group (DM,n=40,UACR<30 mg/gCr),microalbuminuria group (DN 1,n=50,UACR-30~300 mg/gCr) and clinicoalbuminuria group (DN 2,n=30,UACR>300 mg/gCr).Urine exosome-interferon-gamma (IFN-γ) and exosome-interleukin 4 (IL-4) levels were determined by enzyme-linked immunosorbent assay (ELISA).Multiple stepwise linear regression was used to analyze the correlation of exosome-IFN-γ/IL-4 with glycated hemoglobin (HbA1c),cholesterol (CH),UACR,Scr and BUN.Results Th1/Th2 ratio in DM,DN1,DN2 groups was significantly higher than that in healthy group (0.8089±0.2458,0.8993 ±0.3515,0.8571±0.2470 vs 0.6198±0.1769,all P<0.01).Correlation analysis showed that urinary exosomeIFN-γ/IL-4 ratio was positively correlated with UACR (r=0.213,P=0.015) and BUN (r=0.292,P=0.001).Multiple stepwise linear regression analysis showed that BUN was independent determinants for exosome-IFN-γ/IL-4 (β=0.246,P=0.006).Conclusion The imbalance of urinary exosomeTh1/Th2 is correlated with DN,which may play an important role in the pathogenesis of DN.