[目的]探索miR-217与NRP1在奥沙利铂耐药性胃癌细胞中的作用机制。[方法]将MGC803/L-OHP细胞分为4组:miR NC组、miR-217 mimic组、si NC组与si NRP1组。通过实时荧光定量PCR实验和蛋白免疫印迹实验检测耐药和非耐药细胞中miR-217和NRP1...[目的]探索miR-217与NRP1在奥沙利铂耐药性胃癌细胞中的作用机制。[方法]将MGC803/L-OHP细胞分为4组:miR NC组、miR-217 mimic组、si NC组与si NRP1组。通过实时荧光定量PCR实验和蛋白免疫印迹实验检测耐药和非耐药细胞中miR-217和NRP1的表达水平;通过MTT实验分析MGC803/L-OHP细胞的增殖能力;通过Transwell实验分析MGC803/L-OHP细胞的转移活性;通过流式细胞术分析MGC803/L-OHP细胞的凋亡率;通过荧光素酶报告基因实验分析MGC803/L-OHP细胞中miR-217与NRP1的靶向关系。[结果]上调miR-217或抑制NRP1能够抑制非耐药MGC803细胞生长。与MGC803细胞比较,MGC803/L-OHP细胞中的miR-217表达水平降低(0.79±0.08 vs 0.21±0.02;P<0.05),NRP1表达水平增加(0.41±0.09 vs 0.78±0.12;P<0.05)。与miR NC组比较,miR-217 mimic组的MGC803/L-OHP细胞的增殖活性降低、侵袭能力降低、凋亡率增加(2.37±0.23 vs 25.38±5.12;P<0.05)。与si NC组比较,si NRP1组的MGC803/L-OHP细胞的增殖活性降低、侵袭能力降低、凋亡率增加(4.67±0.83 vs 26.17±7.19;P<0.05)。荧光素酶活性测定显示,和miR NC组比较,在NRP1-WT组中转染miR-217 mimic后荧光素酶活性显著降低(0.87±0.03 vs 0.26±0.02;P<0.05)。[结论]在奥沙利铂耐药性胃癌细胞中,miR-217低表达而NRP1高表达,并且miR-217能够靶向抑制NRP1的表达。通过上调miR-217或抑制NRP1的表达能够减弱胃癌细胞对奥沙利铂的耐药性。展开更多
Group 3 innate lymphoid cells(ILC3s)control tissue homeostasis and orchestrate mucosal inflammation;however,the precise mechanisms governing ILC3 activity are fully understood.Here,we identified the transmembrane prot...Group 3 innate lymphoid cells(ILC3s)control tissue homeostasis and orchestrate mucosal inflammation;however,the precise mechanisms governing ILC3 activity are fully understood.Here,we identified the transmembrane protein neuropilin-1(NRP1)as a positive regulator of interleukin(IL)-17-producing ILC3s in the intestine.NRP1 was markedly upregulated in intestinal mucosal biopsies from patients with inflammatory bowel disease(IBD)compared with healthy controls.Genetic deficiency of NRP1 reduces the frequency of ILC3s in the gut and impairs their production of IL-17A in an NF-κB signaling-dependent and cell-intrinsic manner.The diminished IL-17A production in ILC3s altered the composition of the microbiota and improved the outcome of dextran sodium sulfate(DSS)-induced colitis.Furthermore,pharmacological inhibition of NRP1 with EG00229 alleviated the severity of colitis.These observations demonstrated the critical role of NRP1 in the control of intestinal ILC3s,suggesting that NRP1 is a potential therapeutic target for IBD.展开更多
文摘[目的]探索miR-217与NRP1在奥沙利铂耐药性胃癌细胞中的作用机制。[方法]将MGC803/L-OHP细胞分为4组:miR NC组、miR-217 mimic组、si NC组与si NRP1组。通过实时荧光定量PCR实验和蛋白免疫印迹实验检测耐药和非耐药细胞中miR-217和NRP1的表达水平;通过MTT实验分析MGC803/L-OHP细胞的增殖能力;通过Transwell实验分析MGC803/L-OHP细胞的转移活性;通过流式细胞术分析MGC803/L-OHP细胞的凋亡率;通过荧光素酶报告基因实验分析MGC803/L-OHP细胞中miR-217与NRP1的靶向关系。[结果]上调miR-217或抑制NRP1能够抑制非耐药MGC803细胞生长。与MGC803细胞比较,MGC803/L-OHP细胞中的miR-217表达水平降低(0.79±0.08 vs 0.21±0.02;P<0.05),NRP1表达水平增加(0.41±0.09 vs 0.78±0.12;P<0.05)。与miR NC组比较,miR-217 mimic组的MGC803/L-OHP细胞的增殖活性降低、侵袭能力降低、凋亡率增加(2.37±0.23 vs 25.38±5.12;P<0.05)。与si NC组比较,si NRP1组的MGC803/L-OHP细胞的增殖活性降低、侵袭能力降低、凋亡率增加(4.67±0.83 vs 26.17±7.19;P<0.05)。荧光素酶活性测定显示,和miR NC组比较,在NRP1-WT组中转染miR-217 mimic后荧光素酶活性显著降低(0.87±0.03 vs 0.26±0.02;P<0.05)。[结论]在奥沙利铂耐药性胃癌细胞中,miR-217低表达而NRP1高表达,并且miR-217能够靶向抑制NRP1的表达。通过上调miR-217或抑制NRP1的表达能够减弱胃癌细胞对奥沙利铂的耐药性。
基金upported by the following grants:National Natural Science Foundation of China(No.81925018,82130049,82430055,to J.Z.82321001 to Y.Y.,and 82225015 to Q.L.)+1 种基金supported by the National Key Research and Development Project of China(2021ZD0202400,to Q.L.)the New Cornerstone Science Foundation through the XPLORER PRIZE(to Q.L.).
文摘Group 3 innate lymphoid cells(ILC3s)control tissue homeostasis and orchestrate mucosal inflammation;however,the precise mechanisms governing ILC3 activity are fully understood.Here,we identified the transmembrane protein neuropilin-1(NRP1)as a positive regulator of interleukin(IL)-17-producing ILC3s in the intestine.NRP1 was markedly upregulated in intestinal mucosal biopsies from patients with inflammatory bowel disease(IBD)compared with healthy controls.Genetic deficiency of NRP1 reduces the frequency of ILC3s in the gut and impairs their production of IL-17A in an NF-κB signaling-dependent and cell-intrinsic manner.The diminished IL-17A production in ILC3s altered the composition of the microbiota and improved the outcome of dextran sodium sulfate(DSS)-induced colitis.Furthermore,pharmacological inhibition of NRP1 with EG00229 alleviated the severity of colitis.These observations demonstrated the critical role of NRP1 in the control of intestinal ILC3s,suggesting that NRP1 is a potential therapeutic target for IBD.
