Objective:To investigate the effects of exosomes from human T lymphocyte line(H9)treated with HIV envelope protein gp120 on macrophage polarization.Methods:The viability of gp120-treated H9 T lymphocytes was as-sessed...Objective:To investigate the effects of exosomes from human T lymphocyte line(H9)treated with HIV envelope protein gp120 on macrophage polarization.Methods:The viability of gp120-treated H9 T lymphocytes was as-sessed using the CCK8 assay.Inflammatory cytokine levels in the supernatant of H9 cells were determined by ELISA.Exosome characteristics were identified through electron microscopy and Western blot experiments.PHK67 staining was employed to observe the uptake of exosomes by macrophages.Finally,macrophage polarization markers were detected using ELISA and immuno-fluorescence to analyze the impact of gp120-treated T cell exosomes on macrophage polarization.Results:HIV gp120 protein inhibited the proliferation of human T lymphocytes H9 and promoted the release of inflammatory cytokines.Exosomes from H9 T lymphocytes were successfully isolated,displaying a cup-shaped membranous structure under electron microscopy and overexpressing marker proteins CD9,CD63,and CD81.PHK67 staining results indicated that exosomes from H9 cells could be internalized by macrophages.Exosomes from gp120-treated H9 cells promoted the polarization of macrophages towards the M2 pheno-type.Conclusion:Exosomes secreted by human T lymphocytes H9 treated with HIV envelope protein gp120 can promote M2 polarization of macrophages,suggesting a potential novel mechanism of gp120 in immune modulation.展开更多
Objective:To investigate the effect of mir-3168 on the malignant transformation and cisplatin resistance of AGS and AGS/DDP gastric cancer cells,and to verify its target gene.Methods:The expression of mir-3168 in AGS ...Objective:To investigate the effect of mir-3168 on the malignant transformation and cisplatin resistance of AGS and AGS/DDP gastric cancer cells,and to verify its target gene.Methods:The expression of mir-3168 in AGS and AGS/DDP gastric cancer cells was detected by qPCR,and mir-3168 mimic,inhibitor and negative control were synthesized.They were transfected into AGS and AGS/DDP gastric cancer cells,respectively.The expression of mir-3168 and TP53 mRNA was detected by qPCR.Cell viability was detected by CCK8 under gradient cisplatin treatment and non treatment,apoptosis was detected by flow cytometry,cell invasion was detected by Transwell,and TP53 protein expression was detected by western blot,The database predicted the binding sites of mir-3168 and TP53.According to the binding sites,the double luciferase experiment was used to verify the binding of mir-3168 and TP53.Results:Compared with cisplatin sensitive gastric cancer cell AGS,mir-3168 was significantly overexpressed in cisplatin resistant gastric cancer cell AGS/DDP;mir-3168 mimic promotes cisplatin resistance,proliferation and invasion of AGS and AGS/DDP gastric cancer cells,and inhibits apoptosis of AGS and AGS/DDP gastric cancer cells;mir-3168 inhibitor inhibits cisplatin resistance,proliferation and invasion of AGS and AGS/DDP gastric cancer cells,and promotes apoptosis of AGS and AGS/DDP gastric cancer cells;mir-3168 mimic inhibits the expression of TP53 mRNA and protein,and mir-3168 inhibitor promotes the expression of TP53 mRNA and protein;Targetscan database predicted that there was a binding point between mir-3168 and TP53,and the double luciferase experiment suggested that mir-3168 was bound to TP53 through the predicted binding site.Conclusion:mir-3168 may promote the malignant transformation of AGS and AGS/DDP gastric cancer cells and cisplatin resistance by targeting TP53.展开更多
基金National Natural Science Foundation Project(No.81960303)Baise City Scientific Research and Technology Development Plan Project(No.Encyclopedia 20213301,Encyclopedia 20213242,Encyclopedia 20194701)+3 种基金Guangxi Health Commission self funded research project(No.Z20190202,Z20211114)Guangxi University Young and Middle School Teachers'Basic Research Ability Enhancement Project(No.2021KY0538)2022 Traditional Chinese Medicine Self funded Research Project(No.GXZYL20220304)Open Project of Guangxi Key Laboratory of Molecular Pathology of Hepatobiliary Diseases(No.GXZDSYS-009)。
文摘Objective:To investigate the effects of exosomes from human T lymphocyte line(H9)treated with HIV envelope protein gp120 on macrophage polarization.Methods:The viability of gp120-treated H9 T lymphocytes was as-sessed using the CCK8 assay.Inflammatory cytokine levels in the supernatant of H9 cells were determined by ELISA.Exosome characteristics were identified through electron microscopy and Western blot experiments.PHK67 staining was employed to observe the uptake of exosomes by macrophages.Finally,macrophage polarization markers were detected using ELISA and immuno-fluorescence to analyze the impact of gp120-treated T cell exosomes on macrophage polarization.Results:HIV gp120 protein inhibited the proliferation of human T lymphocytes H9 and promoted the release of inflammatory cytokines.Exosomes from H9 T lymphocytes were successfully isolated,displaying a cup-shaped membranous structure under electron microscopy and overexpressing marker proteins CD9,CD63,and CD81.PHK67 staining results indicated that exosomes from H9 cells could be internalized by macrophages.Exosomes from gp120-treated H9 cells promoted the polarization of macrophages towards the M2 pheno-type.Conclusion:Exosomes secreted by human T lymphocytes H9 treated with HIV envelope protein gp120 can promote M2 polarization of macrophages,suggesting a potential novel mechanism of gp120 in immune modulation.
基金This study was supported by National Natural Science Foundation of China(81960303)Youjiang Medical College for Nationalities Affiliated Hospital(R202011710)+6 种基金Youjiang Medical College for Nationalities Affiliated Hospital,Youjiang Key Talents Research Project(Y20212603)Guangxi Key Laboratory of Molecular Pathology of Hepatobiliary Diseases,Affiliated Hospital of Youjiang Medical College for Nationalities(GxZDSYs-009)Scientific Research and Technology Development Program of Baise City(Baike 20213301)Scientific Research and Technology Development Program of Baise City(Baike 20213242)Self-funded research project of Health Commission of Guangxi Zhuang Autonomous Region(20190953)Self-funded research Project of Administration of Traditional Chinese Medicine of Guangxi Zhuang Autonomous Region(GXZYL20220304)Guangxi University Young and Middle-aged Teachers Basic Research Ability Improvement Project(2021KY0538)。
文摘Objective:To investigate the effect of mir-3168 on the malignant transformation and cisplatin resistance of AGS and AGS/DDP gastric cancer cells,and to verify its target gene.Methods:The expression of mir-3168 in AGS and AGS/DDP gastric cancer cells was detected by qPCR,and mir-3168 mimic,inhibitor and negative control were synthesized.They were transfected into AGS and AGS/DDP gastric cancer cells,respectively.The expression of mir-3168 and TP53 mRNA was detected by qPCR.Cell viability was detected by CCK8 under gradient cisplatin treatment and non treatment,apoptosis was detected by flow cytometry,cell invasion was detected by Transwell,and TP53 protein expression was detected by western blot,The database predicted the binding sites of mir-3168 and TP53.According to the binding sites,the double luciferase experiment was used to verify the binding of mir-3168 and TP53.Results:Compared with cisplatin sensitive gastric cancer cell AGS,mir-3168 was significantly overexpressed in cisplatin resistant gastric cancer cell AGS/DDP;mir-3168 mimic promotes cisplatin resistance,proliferation and invasion of AGS and AGS/DDP gastric cancer cells,and inhibits apoptosis of AGS and AGS/DDP gastric cancer cells;mir-3168 inhibitor inhibits cisplatin resistance,proliferation and invasion of AGS and AGS/DDP gastric cancer cells,and promotes apoptosis of AGS and AGS/DDP gastric cancer cells;mir-3168 mimic inhibits the expression of TP53 mRNA and protein,and mir-3168 inhibitor promotes the expression of TP53 mRNA and protein;Targetscan database predicted that there was a binding point between mir-3168 and TP53,and the double luciferase experiment suggested that mir-3168 was bound to TP53 through the predicted binding site.Conclusion:mir-3168 may promote the malignant transformation of AGS and AGS/DDP gastric cancer cells and cisplatin resistance by targeting TP53.
