OBJECTIVE To explore whether MDM2 transfection can alter the MDM2-p53 autoregulatory feedback loop so as to change the sensitivity of ovarian cancer cell lines to cisplatin. METHODS The ovarian cancer cell line A2780 ...OBJECTIVE To explore whether MDM2 transfection can alter the MDM2-p53 autoregulatory feedback loop so as to change the sensitivity of ovarian cancer cell lines to cisplatin. METHODS The ovarian cancer cell line A2780 expressing wild-type P53 and the ovarian cancer cell line SKOV-3 with the p53 null type were stably transfected with pCMV-MDM2 or pCMV as a control. The blocked expression of P53 was determined by Western blots. Cytotoxicity was assessed using the MTT assay and the trypan blue exclusion assay. Flow cytometry was used to detect changes in the cell cycle and removal of platinum -DNA adducts was measured by atomic absorption spec-troscopy. RESULTS (1) Parental A2780 and A2780-V cells (IC50= 15.14±1.39 μmol) have similar cisplatin sensitivities, whereas sensitivity to cisplatin in A2780-M cells (IC50=7.98±1.32 μmol) was 2 to 3 fold greater (P=0.001). The trypan blue exclusion assay demonstrated that cisplatin killed a higher percentage of A2780-M cells compared to A2780-V cells. There was no significant change following MDM2 transfection in SKOV-3 cells. (2) After cisplatin treatment, A2780-M cells showed a pronounced S-phase arrest, however, A2780 cells with the intact wild-type P53, arrested primarily at the G2/M transition. (3) Platinum uptake was similar for all of the A2780 cell lines after ciaplatin treatment, but the removal of plat-inum-DNA adducts was reduced in the A2780-M cells compared with A2780-V cells. CONCLUSION MDM2 increases cisplatin cytotoxicity in ovarian cancer cells by blocking the expression of p53 through the MDM2-p53 autoregulatory feedback loop.展开更多
OBJECTIVE To investigate apoptosis induced by Bax in hepatocellular carcinoma cells and to examine the results of 2 different routes for in vivo gene delivery. METHODS The anti-hepatocellular carcinoma activity of the...OBJECTIVE To investigate apoptosis induced by Bax in hepatocellular carcinoma cells and to examine the results of 2 different routes for in vivo gene delivery. METHODS The anti-hepatocellular carcinoma activity of the Bax gene transferred to the human hepatocellular carcinoma QGY7703 cell line was examined. In addition the Bax gene was transferred in vivo in mice via the caudal vein or hepatic artery to investigate the differences in target organ and non-target organ transfection. RESULTS 1）The Bax gene mediated by a binary adenoviral vector system induced apoptosis in the human hepatic carcinoma QFY7703 cell line. The cell apoptotic rate in the experimental group （Bax） was 50.2±6.9% but only 32.1 ± 9.7% in the Ad/CMV-p53 group, showing that the Bax-apoptotic rate was significantly higher than the control group. 2） LacZ expression was higher in the target organ （liver） when given through the hepatic artery than through the tail vein. In contrast, LacZ expression in the nontarget organs was higher if given through the tail vein compared to the hepatic artery. CONCLUSION Superselective hepatic artery delivery with Bax gene therapy is safe, specific, effective and has low toxicity. This study provided the basis for Bax-gene therapy via the hepatic artery in vivo.展开更多
The aim of this study was to evaluate the performance of an assay using dried plasma spot(DPS)and dried blood spot(DBS)samples for the serological detection of anti-hepatitis C virus(HCV)antibodies.Between January and...The aim of this study was to evaluate the performance of an assay using dried plasma spot(DPS)and dried blood spot(DBS)samples for the serological detection of anti-hepatitis C virus(HCV)antibodies.Between January and July 2019,plasma,DPS and DBS specimens were collected from individuals at high-risk for HCV infection.Samples were tested for anti-HCV by ELISA,and the performance of DPS and DBS specimens was examined using results from the plasma testing,as the standard.Blood samples were collected from 329 persons,including 129 men who have sex with men and 200 intravenous drug users.Results from the plasma testing indicated that 118 samples(59.0%)were HCV positive.Data from the DPS sample testing showed sensitivity as 99.2%(95%confidence interval[CI]:0.95-1.00)and specificity as 100%(95%CI:0.98-1.00)for HCV detection,with Kappa of 99.3%(95%CI:0.98-1.00)while in DBS sample testing the sensitivity as 98.3%(95%CI:0.93-1.00)and specificity as 100%(95%CI:0.98-1.00),with Kappa of 98.7%(95%CI:0.97-1.00),respectively.Spearman’s correlation coefficients for the comparisons between plasma and DPS specimen,plasma and DBS specimens,DPS and DBS specimens were 0.857,0.750,and 0.739,respectively.Compared with the results in plasma,1 sample was not detected using the DPS specimens,and 2 samples were failed for the positive detection,using the DBS specimens.Both DPS and DBS samples were promising alternatives to plasma,for the detection of anti-HCV antibodies.展开更多
文摘OBJECTIVE To explore whether MDM2 transfection can alter the MDM2-p53 autoregulatory feedback loop so as to change the sensitivity of ovarian cancer cell lines to cisplatin. METHODS The ovarian cancer cell line A2780 expressing wild-type P53 and the ovarian cancer cell line SKOV-3 with the p53 null type were stably transfected with pCMV-MDM2 or pCMV as a control. The blocked expression of P53 was determined by Western blots. Cytotoxicity was assessed using the MTT assay and the trypan blue exclusion assay. Flow cytometry was used to detect changes in the cell cycle and removal of platinum -DNA adducts was measured by atomic absorption spec-troscopy. RESULTS (1) Parental A2780 and A2780-V cells (IC50= 15.14±1.39 μmol) have similar cisplatin sensitivities, whereas sensitivity to cisplatin in A2780-M cells (IC50=7.98±1.32 μmol) was 2 to 3 fold greater (P=0.001). The trypan blue exclusion assay demonstrated that cisplatin killed a higher percentage of A2780-M cells compared to A2780-V cells. There was no significant change following MDM2 transfection in SKOV-3 cells. (2) After cisplatin treatment, A2780-M cells showed a pronounced S-phase arrest, however, A2780 cells with the intact wild-type P53, arrested primarily at the G2/M transition. (3) Platinum uptake was similar for all of the A2780 cell lines after ciaplatin treatment, but the removal of plat-inum-DNA adducts was reduced in the A2780-M cells compared with A2780-V cells. CONCLUSION MDM2 increases cisplatin cytotoxicity in ovarian cancer cells by blocking the expression of p53 through the MDM2-p53 autoregulatory feedback loop.
文摘OBJECTIVE To investigate apoptosis induced by Bax in hepatocellular carcinoma cells and to examine the results of 2 different routes for in vivo gene delivery. METHODS The anti-hepatocellular carcinoma activity of the Bax gene transferred to the human hepatocellular carcinoma QGY7703 cell line was examined. In addition the Bax gene was transferred in vivo in mice via the caudal vein or hepatic artery to investigate the differences in target organ and non-target organ transfection. RESULTS 1）The Bax gene mediated by a binary adenoviral vector system induced apoptosis in the human hepatic carcinoma QFY7703 cell line. The cell apoptotic rate in the experimental group （Bax） was 50.2±6.9% but only 32.1 ± 9.7% in the Ad/CMV-p53 group, showing that the Bax-apoptotic rate was significantly higher than the control group. 2） LacZ expression was higher in the target organ （liver） when given through the hepatic artery than through the tail vein. In contrast, LacZ expression in the nontarget organs was higher if given through the tail vein compared to the hepatic artery. CONCLUSION Superselective hepatic artery delivery with Bax gene therapy is safe, specific, effective and has low toxicity. This study provided the basis for Bax-gene therapy via the hepatic artery in vivo.
基金supported by the National Science and Technology Major Project of China in the 13th Five-Year(2017ZX10201101-002-003)
文摘The aim of this study was to evaluate the performance of an assay using dried plasma spot(DPS)and dried blood spot(DBS)samples for the serological detection of anti-hepatitis C virus(HCV)antibodies.Between January and July 2019,plasma,DPS and DBS specimens were collected from individuals at high-risk for HCV infection.Samples were tested for anti-HCV by ELISA,and the performance of DPS and DBS specimens was examined using results from the plasma testing,as the standard.Blood samples were collected from 329 persons,including 129 men who have sex with men and 200 intravenous drug users.Results from the plasma testing indicated that 118 samples(59.0%)were HCV positive.Data from the DPS sample testing showed sensitivity as 99.2%(95%confidence interval[CI]:0.95-1.00)and specificity as 100%(95%CI:0.98-1.00)for HCV detection,with Kappa of 99.3%(95%CI:0.98-1.00)while in DBS sample testing the sensitivity as 98.3%(95%CI:0.93-1.00)and specificity as 100%(95%CI:0.98-1.00),with Kappa of 98.7%(95%CI:0.97-1.00),respectively.Spearman’s correlation coefficients for the comparisons between plasma and DPS specimen,plasma and DBS specimens,DPS and DBS specimens were 0.857,0.750,and 0.739,respectively.Compared with the results in plasma,1 sample was not detected using the DPS specimens,and 2 samples were failed for the positive detection,using the DBS specimens.Both DPS and DBS samples were promising alternatives to plasma,for the detection of anti-HCV antibodies.
