Objective： In this study, we examine the effects of recombinant adenovirus-p53 （rAd-p53） on the pancreatic carcinoma cell line SW1990. Specifically, we determine if expression of rAd-p53 sensitizes these cells to r...Objective： In this study, we examine the effects of recombinant adenovirus-p53 （rAd-p53） on the pancreatic carcinoma cell line SW1990. Specifically, we determine if expression of rAd-p53 sensitizes these cells to radiation. Methods： Following transfection of SW1990 cells with rAd-p53, we measured expression of P53, P21 and Bax by immunocytochemistry. Both transfected and control cell lines were irradiated with a range of doses, and the survival fractions （SF） were calculated. Dose survival cttrves were constructed and modeled for comparison. Results： Transfection of SW1990 cells with rAd-p53 resulted in increased expression of P53, P21 and Bax in a time-dependent manner. At 96 h after transfection, 89.92% of cells expressed P53, 56.8% expressed P21, and 76.50% expressed Bax. The SF following radiation was lower in the rAd-p53 transfected cells compared to the control cells, suggesting that rAd-p53 sensitizes SW1990 cells to radiation （Do for the experimental and control groups was 2.199 and 2.462, respectively）. Conclusions： Use of the adenoviral vector is an effective means of transfecting SW1990 cells with wild-type P53, and this sensitizes the cell line to irradiation. This work suggests that combining rAd-p53 with radiation therapy in pancreatic cancer may be therapeutically beneficial.展开更多
Objective： We previously found that hUTP14a binds P53 and promotes P53 degradation. However, if hUTP14a is a downstream gene of P53 remains to be determined. This study aimed to identify the promoter of h UTP14a and ...Objective： We previously found that hUTP14a binds P53 and promotes P53 degradation. However, if hUTP14a is a downstream gene of P53 remains to be determined. This study aimed to identify the promoter of h UTP14a and investigate if h UTP14a is regulated by P53. Methods： The hUTPI4a promoter region was cloned into pGL3-Basic-luciferase reporter plasmid to get pGL3-hUTP14a-luc. The reporter plasmid was transfected into 293T cells and luciferase activity was evaluated by the Dual-Luciferase Reporter Assay System. Putative transcription factors were identified through searching Matlnspector Professional and Algorismica i Genetica databases. Either pGL3-hUTPI4aluc or p21 promoter reporter plasmid was co-transfected with increasing dose of p53 plasmid, and luciferase activity was evaluated. A series of deletion constructs of pGL3-hUTP14a-luc were constructed and minimal promoter region of hUTP14a was determined. Differences of the lnciferase activities between different groups were assessed by statistical analysis. Results： The hUTP14a gene promoter reporter construct was correctly cloned and was demonstrated to possess promoter activity. The transcription of hUTP14a was not regulated by P53. The minimal promoter region of h UTP14a gene is located between -203 to -100 of the transcription initiation site. Conclusion： Unlike other P53-interacting proteins such as MDM2, Pirh2 and Cop I which promote P53 degradation and whose transcriptions are regulated by P53, does not hUTP14a transcription form a regulation feedback loop with P 53.展开更多
To evaluate the effects of adenovirus (Ad) - mediated transfer of p5 3and p16 on hum an bladder cancer cells EJ,EJwere transfected with Ad- p5 3and Ad- p16 . Cell growth,m orphologi- cal change,cell cycle,apoptosis ...To evaluate the effects of adenovirus (Ad) - mediated transfer of p5 3and p16 on hum an bladder cancer cells EJ,EJwere transfected with Ad- p5 3and Ad- p16 . Cell growth,m orphologi- cal change,cell cycle,apoptosis were measured using MTT assay,flow cytom etry,cloning form a- tion,im munocytochemical assays.Ad- p16 or Ad- p5 3alone could inhibit the proliferating activity of EJcells in vitro.Ad- p5 3could induce apoptosis of partial EJcells.G1arrest was observed72 h after infection with Ad- p16 ,but apoptosis was not obvious.The transfer of Ad- p16 and Ad- p5 3 could significantly inhibit the growth of EJcells,decrease the cloning formation rate and induce apoptosis of large num ber of EJcells. The occurrence time of subcutaneous tumor was delayed and the tum or volume in 4 weeks was dim inished by using Ad- p5 3com bined with Ad- p16 and the dif- ference was significant com pared with using Ad- p5 3or Ad- p16 alone.It was suggested that the transfer of wild- type p5 3and p16 could significantly inhibit the growth of human bladder cancer in vitro and in vivo.展开更多
目的 探讨人细胞色素P450家族成员3A5(cytochrome P450 family member 3A5, CYP3A5)编码基因变异对糖皮质激素的代谢影响以及与垂体前叶功能减退的关系。方法 回顾性选取2021年3月青岛大学附属医院肾病科的一例病理类型为膜性肾病合并Ig...目的 探讨人细胞色素P450家族成员3A5(cytochrome P450 family member 3A5, CYP3A5)编码基因变异对糖皮质激素的代谢影响以及与垂体前叶功能减退的关系。方法 回顾性选取2021年3月青岛大学附属医院肾病科的一例病理类型为膜性肾病合并IgA肾病的肾病综合征患者的临床资料,并对该病进行文献复习。结果 该患者给予常规剂量他克莫司治疗后,检测其他克莫司血药浓度异常增高,发现患者他克莫司代谢酶CYP3A5编码基因存在纯合突变,导致他克莫司蓄积。同时发现该患者垂体前叶功能减退,考虑患者垂体前叶功能减退与糖皮质激素蓄积并反馈抑制垂体前叶功能有关。经过调整糖皮质激素及他克莫司用量,监测患者甲功三项,指标均逐渐好转。结论 CYP3A5基因多态性引起的CYP3A5代谢酶慢代谢型变异,会导致其代谢药物的蓄积,尤其是糖皮质激素,进而引起包括垂体前叶功能减退在内的一系列临床不良结局。展开更多
基金supported by Fujian Province Natural Science Foundation(No.2012J05139)Beijing Municipal Science & Technology Commission(No.Z111107058811021)
文摘Objective： In this study, we examine the effects of recombinant adenovirus-p53 （rAd-p53） on the pancreatic carcinoma cell line SW1990. Specifically, we determine if expression of rAd-p53 sensitizes these cells to radiation. Methods： Following transfection of SW1990 cells with rAd-p53, we measured expression of P53, P21 and Bax by immunocytochemistry. Both transfected and control cell lines were irradiated with a range of doses, and the survival fractions （SF） were calculated. Dose survival cttrves were constructed and modeled for comparison. Results： Transfection of SW1990 cells with rAd-p53 resulted in increased expression of P53, P21 and Bax in a time-dependent manner. At 96 h after transfection, 89.92% of cells expressed P53, 56.8% expressed P21, and 76.50% expressed Bax. The SF following radiation was lower in the rAd-p53 transfected cells compared to the control cells, suggesting that rAd-p53 sensitizes SW1990 cells to radiation （Do for the experimental and control groups was 2.199 and 2.462, respectively）. Conclusions： Use of the adenoviral vector is an effective means of transfecting SW1990 cells with wild-type P53, and this sensitizes the cell line to irradiation. This work suggests that combining rAd-p53 with radiation therapy in pancreatic cancer may be therapeutically beneficial.
基金supported by grants from Beijing Municipal Natural Science Foundation (Grant No. 7122032)the National Natural Science Foundation of China (Grant No. 81071672)+1 种基金the National Basic Research Program of China (973 program) (Grant No. 2010CB529303)National High Technology Research and Development Program of China (863 Program, Grant No. 2008AA02Z131)
文摘Objective： We previously found that hUTP14a binds P53 and promotes P53 degradation. However, if hUTP14a is a downstream gene of P53 remains to be determined. This study aimed to identify the promoter of h UTP14a and investigate if h UTP14a is regulated by P53. Methods： The hUTPI4a promoter region was cloned into pGL3-Basic-luciferase reporter plasmid to get pGL3-hUTP14a-luc. The reporter plasmid was transfected into 293T cells and luciferase activity was evaluated by the Dual-Luciferase Reporter Assay System. Putative transcription factors were identified through searching Matlnspector Professional and Algorismica i Genetica databases. Either pGL3-hUTPI4aluc or p21 promoter reporter plasmid was co-transfected with increasing dose of p53 plasmid, and luciferase activity was evaluated. A series of deletion constructs of pGL3-hUTP14a-luc were constructed and minimal promoter region of hUTP14a was determined. Differences of the lnciferase activities between different groups were assessed by statistical analysis. Results： The hUTP14a gene promoter reporter construct was correctly cloned and was demonstrated to possess promoter activity. The transcription of hUTP14a was not regulated by P53. The minimal promoter region of h UTP14a gene is located between -203 to -100 of the transcription initiation site. Conclusion： Unlike other P53-interacting proteins such as MDM2, Pirh2 and Cop I which promote P53 degradation and whose transcriptions are regulated by P53, does not hUTP14a transcription form a regulation feedback loop with P 53.
基金Thisprojectwassupported by a grant from 86 3Project ofChina(No.Z2 0 - 0 1- 0 2 )
文摘To evaluate the effects of adenovirus (Ad) - mediated transfer of p5 3and p16 on hum an bladder cancer cells EJ,EJwere transfected with Ad- p5 3and Ad- p16 . Cell growth,m orphologi- cal change,cell cycle,apoptosis were measured using MTT assay,flow cytom etry,cloning form a- tion,im munocytochemical assays.Ad- p16 or Ad- p5 3alone could inhibit the proliferating activity of EJcells in vitro.Ad- p5 3could induce apoptosis of partial EJcells.G1arrest was observed72 h after infection with Ad- p16 ,but apoptosis was not obvious.The transfer of Ad- p16 and Ad- p5 3 could significantly inhibit the growth of EJcells,decrease the cloning formation rate and induce apoptosis of large num ber of EJcells. The occurrence time of subcutaneous tumor was delayed and the tum or volume in 4 weeks was dim inished by using Ad- p5 3com bined with Ad- p16 and the dif- ference was significant com pared with using Ad- p5 3or Ad- p16 alone.It was suggested that the transfer of wild- type p5 3and p16 could significantly inhibit the growth of human bladder cancer in vitro and in vivo.
文摘目的 探讨人细胞色素P450家族成员3A5(cytochrome P450 family member 3A5, CYP3A5)编码基因变异对糖皮质激素的代谢影响以及与垂体前叶功能减退的关系。方法 回顾性选取2021年3月青岛大学附属医院肾病科的一例病理类型为膜性肾病合并IgA肾病的肾病综合征患者的临床资料,并对该病进行文献复习。结果 该患者给予常规剂量他克莫司治疗后,检测其他克莫司血药浓度异常增高,发现患者他克莫司代谢酶CYP3A5编码基因存在纯合突变,导致他克莫司蓄积。同时发现该患者垂体前叶功能减退,考虑患者垂体前叶功能减退与糖皮质激素蓄积并反馈抑制垂体前叶功能有关。经过调整糖皮质激素及他克莫司用量,监测患者甲功三项,指标均逐渐好转。结论 CYP3A5基因多态性引起的CYP3A5代谢酶慢代谢型变异,会导致其代谢药物的蓄积,尤其是糖皮质激素,进而引起包括垂体前叶功能减退在内的一系列临床不良结局。
