背景与目的:研究发现核糖体蛋白L8(ribosomal protein L8,RPL8)在黑色素瘤中表达能激活黑色素瘤患者外周血单个核细胞增殖并产生白细胞介素2,提示RPL8可能参与抗肿瘤免疫应答,有望成为抗肿瘤治疗的靶点。本研究通过RPL8蛋白负载树突状细...背景与目的:研究发现核糖体蛋白L8(ribosomal protein L8,RPL8)在黑色素瘤中表达能激活黑色素瘤患者外周血单个核细胞增殖并产生白细胞介素2,提示RPL8可能参与抗肿瘤免疫应答,有望成为抗肿瘤治疗的靶点。本研究通过RPL8蛋白负载树突状细胞(dentritic cell,DC),探讨负载RPL8蛋白的DC对黑色素瘤的免疫效应。方法:原核表达RPL8蛋白,纯化后致敏小鼠骨髓来源DC,流式细胞仪检测DC表面标志,MTT法检测细胞毒性T淋巴细胞对小鼠黑色素瘤细胞的杀伤作用;负载RPL8蛋白DC免疫治疗小鼠后,观察肿瘤体积变化及小鼠生存时间。结果:纯化蛋白经蛋白质印迹法(Western blot)分析见约28×103大小的特异性条带;DC经RPL8及细菌脂多糖(Lipoplysaccharide,LPS)诱导成熟后细胞表面CD11c、CD80、MHC-Ⅰ类、MHC-Ⅱ类分子表达增高,能激活T淋巴细胞,对B16细胞有抑制作用,RPL8-DC组的抑制率在效靶比为30∶1时高达70%,较PBS组和DC组明显增高;负载RPL8蛋白DC免疫治疗小鼠后,肿瘤体积缩小,小鼠的生存期明显延长。结论:负载RPL8的DC对黑色素瘤有生长抑制作用。展开更多
Ribosomal protein L8 is a component of the 60S subunit of the ribosome and is involved in protein synthesis but its role in Drosophila development is not well understood.We depleted L8 through RNA interference (RNAi) ...Ribosomal protein L8 is a component of the 60S subunit of the ribosome and is involved in protein synthesis but its role in Drosophila development is not well understood.We depleted L8 through RNA interference (RNAi) to examine its effects on fly development both in vivo and in vitro.The results demonstrated that L8 RNAi caused embryonic or first-larval lethality,delay of larval development,defects in eye and wing morphology,and dramatically reduced the number of S2 cells.This indicated that L8 plays a crucial role in Drosophila development.Acridine orange staining of the wing discs showed that apoptosis occurred when L8 was depleted,indicating that depletion of L8 is tightly connected to apoptosis.RT-PCR analyses of the transcription level of genes that are known to be key factors in apoptosis (p53,hid,reaper,dark,Dcp-1) and cell cycle regulation (cdc45,MCM3,cyclin B,incenp) in L8-deficient S2 cells,were consistent with their role in apoptosis induction and cell cycle arrest.These results indicate that depletion of L8 strongly impairs Drosophila development,and that this depletion is associated with cell proliferation arrest and apoptosis,in which p53 may play a central role.展开更多
文摘背景与目的:研究发现核糖体蛋白L8(ribosomal protein L8,RPL8)在黑色素瘤中表达能激活黑色素瘤患者外周血单个核细胞增殖并产生白细胞介素2,提示RPL8可能参与抗肿瘤免疫应答,有望成为抗肿瘤治疗的靶点。本研究通过RPL8蛋白负载树突状细胞(dentritic cell,DC),探讨负载RPL8蛋白的DC对黑色素瘤的免疫效应。方法:原核表达RPL8蛋白,纯化后致敏小鼠骨髓来源DC,流式细胞仪检测DC表面标志,MTT法检测细胞毒性T淋巴细胞对小鼠黑色素瘤细胞的杀伤作用;负载RPL8蛋白DC免疫治疗小鼠后,观察肿瘤体积变化及小鼠生存时间。结果:纯化蛋白经蛋白质印迹法(Western blot)分析见约28×103大小的特异性条带;DC经RPL8及细菌脂多糖(Lipoplysaccharide,LPS)诱导成熟后细胞表面CD11c、CD80、MHC-Ⅰ类、MHC-Ⅱ类分子表达增高,能激活T淋巴细胞,对B16细胞有抑制作用,RPL8-DC组的抑制率在效靶比为30∶1时高达70%,较PBS组和DC组明显增高;负载RPL8蛋白DC免疫治疗小鼠后,肿瘤体积缩小,小鼠的生存期明显延长。结论:负载RPL8的DC对黑色素瘤有生长抑制作用。
基金supported by the National Natural Science Foundation of China (Grant No. 30571332)
文摘Ribosomal protein L8 is a component of the 60S subunit of the ribosome and is involved in protein synthesis but its role in Drosophila development is not well understood.We depleted L8 through RNA interference (RNAi) to examine its effects on fly development both in vivo and in vitro.The results demonstrated that L8 RNAi caused embryonic or first-larval lethality,delay of larval development,defects in eye and wing morphology,and dramatically reduced the number of S2 cells.This indicated that L8 plays a crucial role in Drosophila development.Acridine orange staining of the wing discs showed that apoptosis occurred when L8 was depleted,indicating that depletion of L8 is tightly connected to apoptosis.RT-PCR analyses of the transcription level of genes that are known to be key factors in apoptosis (p53,hid,reaper,dark,Dcp-1) and cell cycle regulation (cdc45,MCM3,cyclin B,incenp) in L8-deficient S2 cells,were consistent with their role in apoptosis induction and cell cycle arrest.These results indicate that depletion of L8 strongly impairs Drosophila development,and that this depletion is associated with cell proliferation arrest and apoptosis,in which p53 may play a central role.
