BACKGROUND Gastric carcinoma(GC)is the third most frequent cause of cancer-related death,highlighting the pressing need for novel clinical treatment options.In this regard,microRNAs(miRNAs)have emerged as a promising ...BACKGROUND Gastric carcinoma(GC)is the third most frequent cause of cancer-related death,highlighting the pressing need for novel clinical treatment options.In this regard,microRNAs(miRNAs)have emerged as a promising therapeutic strategy.Studies have shown that miRNAs can regulate related signaling pathways,acting as tumor suppressors or tumor promoters.AIM To explore the effect of miR-204-3p on GC cells.METHODS We measured the expression levels of miR-204-3p in GC cells using quantitative real-time polymerase chain reaction,followed by the delivery of miR-204-3p overexpression and miR-204-3p knockdown vectors into GC cells.CCK-8 was used to detect the effect of miR-204-3p on the proliferation of GC cells,and the colony formation ability of GC cells was detected by the clonal formation assay.The effects of miR-204-3p on GC cell cycle and apoptosis were detected by flow cytometry.The BABL/c nude mouse subcutaneous tumor model using MKN-45 cells was constructed to verify the effect of miR-204-3p on the tumorigenicity of GC cells.Furthermore,the study investigated the effects of miR-204-3p on various proteins related to the MAPK signaling pathway,necroptosis signaling pathway and apoptosis signaling pathway on GC cells using Western blot techniques.RESULTS Firstly,we found that the expression of miR-204-3p in GC was low.When treated with the lentivirus overexpression vector,miR-204-3p expression significantly increased,but the lentivirus knockout vector had no significant effect on miR-204-3p.In vitro experiments confirmed that miR-204-3p overexpression inhibited GC cell viability,promoted cell apoptosis,blocked the cell cycle,and inhibited colony formation ability.In vivo animal experiments confirmed that miR-204-3p overexpression inhibited subcutaneous tumorigenesis ability in BABL/c nude mice.Simultaneously,our results verified that miR-204-3p overexpression can inhibit GC cell proliferation by inhibiting protein expression levels of KRAS and p-ERK1/2 in the MAPK pathway,as well as inhibiting protein expression levels of p-RIP1 and p-MLK1 in the necroptosis pathway to promote the BCL-2/BAX/Caspase-3 apoptosis pathway.CONCLUSION MiR-204-3p overexpression inhibited GC cell proliferation by inhibiting the MAPK pathway and necroptosis pathway to promote apoptosis of GC cells.Thus,miR-204-3p may represent a new potential therapeutic target for GC.展开更多
目的探讨miR-204-5p对脂多糖(LPS)诱导的肺微血管内皮细胞损伤的影响及其可能作用机制。方法采用LPS诱导大鼠肺微血管内皮细胞(PMVEC)建立细胞损伤模型,实验分组:NC组、LPS组、LPS+miR-con组、LPS+miR-204-5p组、LPS+si-con组、LPS+si-T...目的探讨miR-204-5p对脂多糖(LPS)诱导的肺微血管内皮细胞损伤的影响及其可能作用机制。方法采用LPS诱导大鼠肺微血管内皮细胞(PMVEC)建立细胞损伤模型,实验分组:NC组、LPS组、LPS+miR-con组、LPS+miR-204-5p组、LPS+si-con组、LPS+si-TRIB3组、LPS+miR-204-5p+pcDNA组、LPS+miR-204-5p+pcDNA-TRIB3组;MTT法检测细胞活力;qRT-PCR、Western blot检测miR-204-5p、TRIB3表达;流式细胞术检测细胞凋亡;ELISA检测TNF-α、IL-6水平;双荧光素酶实验检测miR-204-5p与TRIB3的靶向关系。结果与NC组比较,LPS组细胞活力、miR-204-5p表达量降低(1.00±0.10比0.43±0.04),细胞凋亡率、TRIB3 mRNA(1.00±0.09 vs 2.13±0.18)和蛋白(0.40±0.04 vs 0.83±0.08)水平、TNF-α、IL-6水平升高(P<0.05);过表达miR-204-5p或敲低TRIB3可升高细胞活力,降低细胞凋亡率和TNF-α、IL-6水平(P<0.05);TRIB3是miR-204-5p的靶基因,上调TRIB3可减弱过表达miR-204-5p对细胞增殖、凋亡和炎症反应的影响。结论miR-204-5p过表达可靶向负调控TRIB3表达促进细胞增殖,抑制细胞凋亡和炎症反应,从而减轻LPS诱导的肺微血管内皮细胞损伤。展开更多
文摘BACKGROUND Gastric carcinoma(GC)is the third most frequent cause of cancer-related death,highlighting the pressing need for novel clinical treatment options.In this regard,microRNAs(miRNAs)have emerged as a promising therapeutic strategy.Studies have shown that miRNAs can regulate related signaling pathways,acting as tumor suppressors or tumor promoters.AIM To explore the effect of miR-204-3p on GC cells.METHODS We measured the expression levels of miR-204-3p in GC cells using quantitative real-time polymerase chain reaction,followed by the delivery of miR-204-3p overexpression and miR-204-3p knockdown vectors into GC cells.CCK-8 was used to detect the effect of miR-204-3p on the proliferation of GC cells,and the colony formation ability of GC cells was detected by the clonal formation assay.The effects of miR-204-3p on GC cell cycle and apoptosis were detected by flow cytometry.The BABL/c nude mouse subcutaneous tumor model using MKN-45 cells was constructed to verify the effect of miR-204-3p on the tumorigenicity of GC cells.Furthermore,the study investigated the effects of miR-204-3p on various proteins related to the MAPK signaling pathway,necroptosis signaling pathway and apoptosis signaling pathway on GC cells using Western blot techniques.RESULTS Firstly,we found that the expression of miR-204-3p in GC was low.When treated with the lentivirus overexpression vector,miR-204-3p expression significantly increased,but the lentivirus knockout vector had no significant effect on miR-204-3p.In vitro experiments confirmed that miR-204-3p overexpression inhibited GC cell viability,promoted cell apoptosis,blocked the cell cycle,and inhibited colony formation ability.In vivo animal experiments confirmed that miR-204-3p overexpression inhibited subcutaneous tumorigenesis ability in BABL/c nude mice.Simultaneously,our results verified that miR-204-3p overexpression can inhibit GC cell proliferation by inhibiting protein expression levels of KRAS and p-ERK1/2 in the MAPK pathway,as well as inhibiting protein expression levels of p-RIP1 and p-MLK1 in the necroptosis pathway to promote the BCL-2/BAX/Caspase-3 apoptosis pathway.CONCLUSION MiR-204-3p overexpression inhibited GC cell proliferation by inhibiting the MAPK pathway and necroptosis pathway to promote apoptosis of GC cells.Thus,miR-204-3p may represent a new potential therapeutic target for GC.
文摘目的探讨miR-204-5p对脂多糖(LPS)诱导的肺微血管内皮细胞损伤的影响及其可能作用机制。方法采用LPS诱导大鼠肺微血管内皮细胞(PMVEC)建立细胞损伤模型,实验分组:NC组、LPS组、LPS+miR-con组、LPS+miR-204-5p组、LPS+si-con组、LPS+si-TRIB3组、LPS+miR-204-5p+pcDNA组、LPS+miR-204-5p+pcDNA-TRIB3组;MTT法检测细胞活力;qRT-PCR、Western blot检测miR-204-5p、TRIB3表达;流式细胞术检测细胞凋亡;ELISA检测TNF-α、IL-6水平;双荧光素酶实验检测miR-204-5p与TRIB3的靶向关系。结果与NC组比较,LPS组细胞活力、miR-204-5p表达量降低(1.00±0.10比0.43±0.04),细胞凋亡率、TRIB3 mRNA(1.00±0.09 vs 2.13±0.18)和蛋白(0.40±0.04 vs 0.83±0.08)水平、TNF-α、IL-6水平升高(P<0.05);过表达miR-204-5p或敲低TRIB3可升高细胞活力,降低细胞凋亡率和TNF-α、IL-6水平(P<0.05);TRIB3是miR-204-5p的靶基因,上调TRIB3可减弱过表达miR-204-5p对细胞增殖、凋亡和炎症反应的影响。结论miR-204-5p过表达可靶向负调控TRIB3表达促进细胞增殖,抑制细胞凋亡和炎症反应,从而减轻LPS诱导的肺微血管内皮细胞损伤。
