[目的]探讨linc01376在鼻咽癌组织与细胞内的表达特征,分析其可能的调控机制。[方法]采用实时荧光定量(qT-PCR)方法检测鼻咽癌组织和细胞中linc01376表达水平;采用CCK-8法、克隆平板形成实验、流式细胞学检测、划痕实验、Transwell侵袭...[目的]探讨linc01376在鼻咽癌组织与细胞内的表达特征,分析其可能的调控机制。[方法]采用实时荧光定量(qT-PCR)方法检测鼻咽癌组织和细胞中linc01376表达水平;采用CCK-8法、克隆平板形成实验、流式细胞学检测、划痕实验、Transwell侵袭实验等检测细胞增殖、迁移和侵袭能力变化;通过Western blot实验测定分子蛋白水平,探索linc01376的下游调控机制。[结果]qT-PCR结果表明,linc01376在鼻咽癌组织中表达明显高于正常鼻咽组织(20.230±1.815 vs 1.281±0.454,P<0.01)。linc01376表达越高,患者分期越晚(χ2=7.670,P<0.05),肿瘤负荷越大(χ2=6.312,P<0.05)。细胞功能学实验显示与对照组相比,沉默linc01376明显抑制鼻咽癌细胞的增殖(5-8F:2.248±0.055 vs 1.790±0.041,t=6.675,P<0.01;CNE2:1.502±0.046 vs 1.012±0.068,t=5.994,P<0.01)、迁移和侵袭能力(5-8F:182.00±8.60 vs 92.67±6.24,t=11.89,P<0.01;CNE2:166.67±8.18 vs 90.67±3.68,t=11.98,P<0.01)。机制研究发现linc01376可通过影响miR-4757调控下游IGF1的表达,最终激活AKT/p-AKT信号通路发挥促进肿瘤进程作用。[结论]linc01376在鼻咽癌进展中发挥重要调控作用,可能成为鼻咽癌靶向治疗的新靶点。展开更多
The mechanisms of Gardeniae Fructus (GF) for anti-hyperglycemic action were demonstrated in streptozotocin (STZ)-diabetic mice. Six hours after single intraperitoneal administration of GF (300 mg/kg) or H2O into 3 hou...The mechanisms of Gardeniae Fructus (GF) for anti-hyperglycemic action were demonstrated in streptozotocin (STZ)-diabetic mice. Six hours after single intraperitoneal administration of GF (300 mg/kg) or H2O into 3 hour-fasted STZ-diabetic mice, glucose and insulin tolerances were assessed by intraperitoneal glucose (1.5 g/kg) tolerance test (IPGTT) and intraperitoneal insulin (0.65 U/kg) tolerance test (IPITT), respectively. Effects of GF on insulin signaling pathways in soleus muscle such as glucose uptake, expression of glucose transporter 4 (GLUT4) in the plasma membrane and phosphorylation of Akt (P-Akt) in cytosolic fraction were examined in STZ-diabetic mice. In IPGTT test, GF significantly accelerated clearance of exogenous glucose and its glucose-lowering action was greater than H2O-treated controlin STZ-diabetic mice. GF also promoted an exogenous glucose-increased insulin level in STZ-diabetic mice. In IPITT test, GF decreased glucose level to the greater extent than H2O-treated control in STZ-diabetic mice. Furthermore, GF significantly decreased high HOMA-IR in STZ-diabetic mice from 21.6 ± 2.4 to 12.4 ± 1.9 (mg/dl × μU/ml). These results implied that GF improved insulin resistance in STZ-diabetic mice. GF increased glucose uptake of soleus muscle 1.5 times greater than H2O-treated control in STZ-diabetic mice. GF enlarged insulin (10 nmol/ml)-increased glucose uptake to 1.8 time-greater. Correspondingly, GF increased expression of GLUT4 in the plasma membrane of soleus muscle to 1.4 time-greater, and P-Akt in the cytosolic fraction of soleus muscle to 1.9 time-greater than those in H2O-treated control. In conclusion, the improvement of GF on insulin resistance is associated with the repair of insulin signaling via P-Akt, GLUT4 and glucose uptake pathway in soleus muscle of STZ-diabetic mice.展开更多
目的研究血管紧张素(1-7)[Ang(1-7)]对糖脂毒性(GLT)孵育的小鼠胰岛素瘤细胞系MIN6β细胞自噬及磷酸化Akt(p-Akt)表达的影响。方法将细胞分为四组:(1)对照组(CON)(2)高糖高脂组(HGHF)(3)Ang(1-7)干预组(4)A779干预组,细胞接种于含15%胎...目的研究血管紧张素(1-7)[Ang(1-7)]对糖脂毒性(GLT)孵育的小鼠胰岛素瘤细胞系MIN6β细胞自噬及磷酸化Akt(p-Akt)表达的影响。方法将细胞分为四组:(1)对照组(CON)(2)高糖高脂组(HGHF)(3)Ang(1-7)干预组(4)A779干预组,细胞接种于含15%胎牛血清及1%链-青霉素溶液的高糖DMEM培养基,置于饱和湿度、37℃、5%CO2培养箱中培养,按上述分组处理后干预12 h。Western-Blot检测自噬相关蛋白Beclin 1和p62以及p-Akt的表达,DAPI染色荧光显微镜下观察细胞凋亡,Cell Counting Kit-8(CCK-8)检测MIN6细胞的凋亡率。结果与CON相比,HGHF组p62及Beclin1蛋白的表达水平均明显增加(p62:1.79±0.57 vs. 1.0,P=0.024;Beclin1:1.53±0.06 vs. 1.0,P=0.000),p-Akt的表达显著降低(0.38±0.06 vs. 1.0,P=0.000),凋亡率明显升高(48.37±8.99 vs. 4.80±0.30,P=0.000);与HGHF相比,Ang(1-7)干预组p62及Beclin1的表达均下降(p62:0.85±0.12 vs. 1.79±0.57,P=0.011;Beclin1:0.90±0.16 vs. 1.53±0.06,P=0.000),p-Akt的表达升高(0.66±0.07 vs. 0.38±0.06,P=0.001),凋亡率下降(20.63±6.35 vs. 48.37±8.99,P=0.001);与Ang(1-7)干预组相比,A779干预组p62及Beclin的表达均增加(p62:1.55±0.38 vs. 0.85±0.12,P=0.002;Beclin1:1.28±0.09 vs. 0.90±0.16,P=0.039),p-Akt的表达降低(0.41±0.10 vs. 0.66±0.07,P=0.003),凋亡率明显升高(33.43±6.15 vs. 20.63±6.35,P=0.038)。结论 GLT通过减少p-Akt的表达诱导胰岛β细胞自噬和凋亡,Ang(1-7)可介导Akt活化抑制GLT诱导的β细胞自噬,减少GLT诱导的β细胞凋亡。展开更多
文摘[目的]探讨linc01376在鼻咽癌组织与细胞内的表达特征,分析其可能的调控机制。[方法]采用实时荧光定量(qT-PCR)方法检测鼻咽癌组织和细胞中linc01376表达水平;采用CCK-8法、克隆平板形成实验、流式细胞学检测、划痕实验、Transwell侵袭实验等检测细胞增殖、迁移和侵袭能力变化;通过Western blot实验测定分子蛋白水平,探索linc01376的下游调控机制。[结果]qT-PCR结果表明,linc01376在鼻咽癌组织中表达明显高于正常鼻咽组织(20.230±1.815 vs 1.281±0.454,P<0.01)。linc01376表达越高,患者分期越晚(χ2=7.670,P<0.05),肿瘤负荷越大(χ2=6.312,P<0.05)。细胞功能学实验显示与对照组相比,沉默linc01376明显抑制鼻咽癌细胞的增殖(5-8F:2.248±0.055 vs 1.790±0.041,t=6.675,P<0.01;CNE2:1.502±0.046 vs 1.012±0.068,t=5.994,P<0.01)、迁移和侵袭能力(5-8F:182.00±8.60 vs 92.67±6.24,t=11.89,P<0.01;CNE2:166.67±8.18 vs 90.67±3.68,t=11.98,P<0.01)。机制研究发现linc01376可通过影响miR-4757调控下游IGF1的表达,最终激活AKT/p-AKT信号通路发挥促进肿瘤进程作用。[结论]linc01376在鼻咽癌进展中发挥重要调控作用,可能成为鼻咽癌靶向治疗的新靶点。
文摘The mechanisms of Gardeniae Fructus (GF) for anti-hyperglycemic action were demonstrated in streptozotocin (STZ)-diabetic mice. Six hours after single intraperitoneal administration of GF (300 mg/kg) or H2O into 3 hour-fasted STZ-diabetic mice, glucose and insulin tolerances were assessed by intraperitoneal glucose (1.5 g/kg) tolerance test (IPGTT) and intraperitoneal insulin (0.65 U/kg) tolerance test (IPITT), respectively. Effects of GF on insulin signaling pathways in soleus muscle such as glucose uptake, expression of glucose transporter 4 (GLUT4) in the plasma membrane and phosphorylation of Akt (P-Akt) in cytosolic fraction were examined in STZ-diabetic mice. In IPGTT test, GF significantly accelerated clearance of exogenous glucose and its glucose-lowering action was greater than H2O-treated controlin STZ-diabetic mice. GF also promoted an exogenous glucose-increased insulin level in STZ-diabetic mice. In IPITT test, GF decreased glucose level to the greater extent than H2O-treated control in STZ-diabetic mice. Furthermore, GF significantly decreased high HOMA-IR in STZ-diabetic mice from 21.6 ± 2.4 to 12.4 ± 1.9 (mg/dl × μU/ml). These results implied that GF improved insulin resistance in STZ-diabetic mice. GF increased glucose uptake of soleus muscle 1.5 times greater than H2O-treated control in STZ-diabetic mice. GF enlarged insulin (10 nmol/ml)-increased glucose uptake to 1.8 time-greater. Correspondingly, GF increased expression of GLUT4 in the plasma membrane of soleus muscle to 1.4 time-greater, and P-Akt in the cytosolic fraction of soleus muscle to 1.9 time-greater than those in H2O-treated control. In conclusion, the improvement of GF on insulin resistance is associated with the repair of insulin signaling via P-Akt, GLUT4 and glucose uptake pathway in soleus muscle of STZ-diabetic mice.
文摘目的研究血管紧张素(1-7)[Ang(1-7)]对糖脂毒性(GLT)孵育的小鼠胰岛素瘤细胞系MIN6β细胞自噬及磷酸化Akt(p-Akt)表达的影响。方法将细胞分为四组:(1)对照组(CON)(2)高糖高脂组(HGHF)(3)Ang(1-7)干预组(4)A779干预组,细胞接种于含15%胎牛血清及1%链-青霉素溶液的高糖DMEM培养基,置于饱和湿度、37℃、5%CO2培养箱中培养,按上述分组处理后干预12 h。Western-Blot检测自噬相关蛋白Beclin 1和p62以及p-Akt的表达,DAPI染色荧光显微镜下观察细胞凋亡,Cell Counting Kit-8(CCK-8)检测MIN6细胞的凋亡率。结果与CON相比,HGHF组p62及Beclin1蛋白的表达水平均明显增加(p62:1.79±0.57 vs. 1.0,P=0.024;Beclin1:1.53±0.06 vs. 1.0,P=0.000),p-Akt的表达显著降低(0.38±0.06 vs. 1.0,P=0.000),凋亡率明显升高(48.37±8.99 vs. 4.80±0.30,P=0.000);与HGHF相比,Ang(1-7)干预组p62及Beclin1的表达均下降(p62:0.85±0.12 vs. 1.79±0.57,P=0.011;Beclin1:0.90±0.16 vs. 1.53±0.06,P=0.000),p-Akt的表达升高(0.66±0.07 vs. 0.38±0.06,P=0.001),凋亡率下降(20.63±6.35 vs. 48.37±8.99,P=0.001);与Ang(1-7)干预组相比,A779干预组p62及Beclin的表达均增加(p62:1.55±0.38 vs. 0.85±0.12,P=0.002;Beclin1:1.28±0.09 vs. 0.90±0.16,P=0.039),p-Akt的表达降低(0.41±0.10 vs. 0.66±0.07,P=0.003),凋亡率明显升高(33.43±6.15 vs. 20.63±6.35,P=0.038)。结论 GLT通过减少p-Akt的表达诱导胰岛β细胞自噬和凋亡,Ang(1-7)可介导Akt活化抑制GLT诱导的β细胞自噬,减少GLT诱导的β细胞凋亡。
