目的探讨RAD51抑制剂RI-1对MSH2缺陷结直肠癌细胞系的杀伤作用及可能的作用机制。方法 Western blot法筛选MSH2差异表达的结直肠癌细胞系,MTT法检测不同结直肠癌细胞系对RI-1(10、20、30、40或50μmol/L)的敏感性;构建针对MSH2基因的重...目的探讨RAD51抑制剂RI-1对MSH2缺陷结直肠癌细胞系的杀伤作用及可能的作用机制。方法 Western blot法筛选MSH2差异表达的结直肠癌细胞系,MTT法检测不同结直肠癌细胞系对RI-1(10、20、30、40或50μmol/L)的敏感性;构建针对MSH2基因的重组shRNA慢病毒表达载体并感染HT29细胞。RI-1(40μmol/L)处理细胞,流式细胞术检测细胞凋亡的变化;单细胞凝胶电泳实验分析细胞内DNA损伤情况;细胞免疫荧光法检测γ-H2AX foci的形成。结果与对照组相比MSH2缺陷的HCT8细胞有明显细胞凋亡现象(P<0.01);HCT8细胞和HT29 Shmsh2细胞尾部DNA含量、尾长和尾距均较对照组明显增加(P<0.05);HCT8细胞和HT29 Shmsh2细胞内γ-H2AX foci的形成数量较对照组明显增加(P<0.01)。结论 RAD51抑制剂RI-1可能通过增加细胞内DNA损伤参与RI-1对MSH2缺陷肿瘤的杀伤作用。展开更多
The ribosomal proteins are crucial for the maintenance of ribosomal translational efficiency and fidelity.In the study,we characterized the ribosomal protein S30(RPS30)gene from Arachis hypogaea that has been isolated...The ribosomal proteins are crucial for the maintenance of ribosomal translational efficiency and fidelity.In the study,we characterized the ribosomal protein S30(RPS30)gene from Arachis hypogaea that has been isolated through Genefishing analysis during defense responses to Ralstonia solanacearum.The cDNA of RPS 30 contained a 189 base pair(bp)open-reading frame encoding 62 amino acids.The genomic DNA consists of 272 bp containing two exons and one 83 bp intron.The RPS 30 mRNA transcript was mainly expressed in roots and leaves.The expression level of the RPS 30 mRNA transcripts was up-regulated sharply 6 h after bacterial challenge and was 12 times greater than that of the control group.The phylogenetic analysis for genes encoding proteins showed that RPS30 were conserved within dicotyledonous and monocotyledonous plants.d S extremely exceeded d N in all branches of the tree(d N/d S<1.0),indicating that functional constraint have acted on RPS 30 throughout evolution.展开更多
基金Natural Science Funds for Young Scholar of Shandong Academy of Agricultural Sciences,China(No.2015YQN13)Natural Science Foundation of Shandong Province,China(ZR2015YL064)+2 种基金Qingdao Science and Technology Plan Basic Research Project,China(No.12-1-4-11-(1)-jch)China Agricultural Research System(No.CARS-13)Agricultural Scientific and Technological Innovation Project of Shandong Academy of Agricultural Sciences,China(No.CXGC2018E21)
文摘The ribosomal proteins are crucial for the maintenance of ribosomal translational efficiency and fidelity.In the study,we characterized the ribosomal protein S30(RPS30)gene from Arachis hypogaea that has been isolated through Genefishing analysis during defense responses to Ralstonia solanacearum.The cDNA of RPS 30 contained a 189 base pair(bp)open-reading frame encoding 62 amino acids.The genomic DNA consists of 272 bp containing two exons and one 83 bp intron.The RPS 30 mRNA transcript was mainly expressed in roots and leaves.The expression level of the RPS 30 mRNA transcripts was up-regulated sharply 6 h after bacterial challenge and was 12 times greater than that of the control group.The phylogenetic analysis for genes encoding proteins showed that RPS30 were conserved within dicotyledonous and monocotyledonous plants.d S extremely exceeded d N in all branches of the tree(d N/d S<1.0),indicating that functional constraint have acted on RPS 30 throughout evolution.
