目的探讨电压依赖性阴离子通道1(VDAC1)对肺腺癌(LUAD)细胞增殖、迁移和侵袭的调控机制。方法采用生物信息学与实验验证相结合的研究方法,于2023年2月至2024年8月在安徽理工大学医学院中心实验室进行生物信息学分析和细胞学实验验证,并...目的探讨电压依赖性阴离子通道1(VDAC1)对肺腺癌(LUAD)细胞增殖、迁移和侵袭的调控机制。方法采用生物信息学与实验验证相结合的研究方法,于2023年2月至2024年8月在安徽理工大学医学院中心实验室进行生物信息学分析和细胞学实验验证,并利用免疫组化进行临床组织学标本验证,回顾性分析南京医科大学附属淮安第一人民医院5例肺腺癌和癌旁样本。TCGA网络数据库分析了VDAC1在LUAD中的表达模式、预后价值和功能富集情况。通过慢病毒转染建立VDAC1敲低的A549和VDAC1过表达的H1650细胞模型。通过免疫组化法检测了LUAD组织和癌旁组织标本中VDAC1蛋白的表达差异。通过CCK8实验、划痕愈合实验和Transwell实验探讨VDAC1对LUAD增殖、迁移和侵袭能力的影响。通过Western blot检测上皮-间充质转化(EMT)标志物蛋白、细胞周期蛋白依赖性激酶和PI3K/AKT/mTOR信号通路分子的活化水平。结果生物信息学分析揭示VDAC1在LUAD细胞中高表达(P<0.0001),是LUAD的独立危险因素(P<0.0001)。功能富集分析显示PI3K/AKT/mTOR、G2M检查点、P53信号通路被显著富集(P<0.001)。与癌旁对照组织相比,VDAC1蛋白在肺腺癌组织中的表达水平更高。过表达VDAC1能够促进H1650细胞增殖(P<0.0001)、迁移和侵袭(P<0.01),敲低VDAC1能够抑制A549细胞增殖(P<0.0001)、迁移和侵袭(P<0.05)。Western Blot实验显示,与对照组比较,过表达VDAC1的H1650细胞中波形蛋白(1.10±0.11 vs 2.39±0.15,P<0.001)、N-钙黏蛋白(0.94±0.12 vs 2.72±0.06,P<0.001)、CDK1(0.93±0.04 vs 1.53±0.03,P<0.0001)、CDK2(1.04±0.13 vs 2.29±0.06,P<0.001)、CDK4(0.90±0.03 vs 2.00±0.11,P<0.01)、p-PI3K(1.08±0.13 vs 1.85±0.12,P<0.01)、p-AKT(1.03±0.11 vs 1.69±0.06,P<0.001)等表达增高,E-钙黏蛋白表达水平下降(2.18±0.14 vs 0.997±0.11,P<0.001);而敲低VDAC1的A549细胞中波形蛋白(1.70±0.26 vs 0.97±0.09,P<0.05)、N-钙黏蛋白(1.98±0.25 vs 1.03±0.06,P<0.05)、CDK1(1.13±0.03 vs 0.95±0.02,P<0.01)、CDK2(2.29±0.12 vs 0.92±0.10,P<0.001)、CDK4(1.71±0.096 vs 1.12±0.11,P<0.01)、p-PI3K(1.67±0.09 vs 0.97±0.03,P<0.001)、p-AKT(1.53±0.04 vs 1.02±0.03,P<0.0001)等表达下降,E-钙黏蛋白表达水平升高(1.04±0.04 vs 1.85±0.26,P<0.05)。结论VDAC1可能通过PI3K/AKT/mTOR激活EMT和细胞周期蛋白依赖性激酶,进而促进LUAD细胞的增殖、迁移和侵袭。展开更多
Objective:To observe the effects of acupuncture-moxibustion combined with rehabilitation training on limb function and surface electromyography(sEMG)in patients with hemiplegia after stroke(HAS).Methods:Eighty-four HA...Objective:To observe the effects of acupuncture-moxibustion combined with rehabilitation training on limb function and surface electromyography(sEMG)in patients with hemiplegia after stroke(HAS).Methods:Eighty-four HAS patients were divided into a control group and a combined group using the random number table method,with 42 cases in each group.The control group received dynamic-static balance rehabilitation training,and the combined group additionally received Fu Zheng Bu Tu(strengthening the healthy Qi and invigorating the spleen)acupuncture-moxibustion therapy.After 4 weeks of treatment,their clinical efficacy was compared,and they were also observed for changes in limb motor function,muscle spasticity level,balance ability,sEMG indices,and hemorheological parameters,as well as adverse reactions.Results:After treatment,the combined group had a higher total effective rate than the control group(P<0.05).After the intervention,both groups presented significant increases in the upper-and lower-limb Fugl-Meyer assessment(FMA)score,Berg balance scale(BBS)score,maximum voluntary contraction(MVC),root mean square(RMS),and integrated electromyography(iEMG);the combined group had superior improvements compared to the control group(P<0.05).After treatment,the modified Ashworth scale(MAS)scores of the elbow,wrist,knee,and ankle joints,and the levels of plasma viscosity(PV),whole blood viscosity(WBV),and hematocrit(HCT)dropped in both groups and were lower in the combined group than in the control group(P<0.05).Neither group showed significant adverse reactions during the treatment period.Conclusion:Fu Zheng Bu Tu acupuncture-moxibustion plus dynamic-static balance rehabilitation training can effectively alleviate muscle spasticity,improve balance ability,enhance sEMG signal levels,modulate hemodynamics,and promote the recovery of limb motor function in HAS patients.展开更多
文摘目的探讨电压依赖性阴离子通道1(VDAC1)对肺腺癌(LUAD)细胞增殖、迁移和侵袭的调控机制。方法采用生物信息学与实验验证相结合的研究方法,于2023年2月至2024年8月在安徽理工大学医学院中心实验室进行生物信息学分析和细胞学实验验证,并利用免疫组化进行临床组织学标本验证,回顾性分析南京医科大学附属淮安第一人民医院5例肺腺癌和癌旁样本。TCGA网络数据库分析了VDAC1在LUAD中的表达模式、预后价值和功能富集情况。通过慢病毒转染建立VDAC1敲低的A549和VDAC1过表达的H1650细胞模型。通过免疫组化法检测了LUAD组织和癌旁组织标本中VDAC1蛋白的表达差异。通过CCK8实验、划痕愈合实验和Transwell实验探讨VDAC1对LUAD增殖、迁移和侵袭能力的影响。通过Western blot检测上皮-间充质转化(EMT)标志物蛋白、细胞周期蛋白依赖性激酶和PI3K/AKT/mTOR信号通路分子的活化水平。结果生物信息学分析揭示VDAC1在LUAD细胞中高表达(P<0.0001),是LUAD的独立危险因素(P<0.0001)。功能富集分析显示PI3K/AKT/mTOR、G2M检查点、P53信号通路被显著富集(P<0.001)。与癌旁对照组织相比,VDAC1蛋白在肺腺癌组织中的表达水平更高。过表达VDAC1能够促进H1650细胞增殖(P<0.0001)、迁移和侵袭(P<0.01),敲低VDAC1能够抑制A549细胞增殖(P<0.0001)、迁移和侵袭(P<0.05)。Western Blot实验显示,与对照组比较,过表达VDAC1的H1650细胞中波形蛋白(1.10±0.11 vs 2.39±0.15,P<0.001)、N-钙黏蛋白(0.94±0.12 vs 2.72±0.06,P<0.001)、CDK1(0.93±0.04 vs 1.53±0.03,P<0.0001)、CDK2(1.04±0.13 vs 2.29±0.06,P<0.001)、CDK4(0.90±0.03 vs 2.00±0.11,P<0.01)、p-PI3K(1.08±0.13 vs 1.85±0.12,P<0.01)、p-AKT(1.03±0.11 vs 1.69±0.06,P<0.001)等表达增高,E-钙黏蛋白表达水平下降(2.18±0.14 vs 0.997±0.11,P<0.001);而敲低VDAC1的A549细胞中波形蛋白(1.70±0.26 vs 0.97±0.09,P<0.05)、N-钙黏蛋白(1.98±0.25 vs 1.03±0.06,P<0.05)、CDK1(1.13±0.03 vs 0.95±0.02,P<0.01)、CDK2(2.29±0.12 vs 0.92±0.10,P<0.001)、CDK4(1.71±0.096 vs 1.12±0.11,P<0.01)、p-PI3K(1.67±0.09 vs 0.97±0.03,P<0.001)、p-AKT(1.53±0.04 vs 1.02±0.03,P<0.0001)等表达下降,E-钙黏蛋白表达水平升高(1.04±0.04 vs 1.85±0.26,P<0.05)。结论VDAC1可能通过PI3K/AKT/mTOR激活EMT和细胞周期蛋白依赖性激酶,进而促进LUAD细胞的增殖、迁移和侵袭。
基金supported by the Health and Medical Science Research Project of Shaanxi Province(陕西省卫生健康科研项目,No.2021H0217).
文摘Objective:To observe the effects of acupuncture-moxibustion combined with rehabilitation training on limb function and surface electromyography(sEMG)in patients with hemiplegia after stroke(HAS).Methods:Eighty-four HAS patients were divided into a control group and a combined group using the random number table method,with 42 cases in each group.The control group received dynamic-static balance rehabilitation training,and the combined group additionally received Fu Zheng Bu Tu(strengthening the healthy Qi and invigorating the spleen)acupuncture-moxibustion therapy.After 4 weeks of treatment,their clinical efficacy was compared,and they were also observed for changes in limb motor function,muscle spasticity level,balance ability,sEMG indices,and hemorheological parameters,as well as adverse reactions.Results:After treatment,the combined group had a higher total effective rate than the control group(P<0.05).After the intervention,both groups presented significant increases in the upper-and lower-limb Fugl-Meyer assessment(FMA)score,Berg balance scale(BBS)score,maximum voluntary contraction(MVC),root mean square(RMS),and integrated electromyography(iEMG);the combined group had superior improvements compared to the control group(P<0.05).After treatment,the modified Ashworth scale(MAS)scores of the elbow,wrist,knee,and ankle joints,and the levels of plasma viscosity(PV),whole blood viscosity(WBV),and hematocrit(HCT)dropped in both groups and were lower in the combined group than in the control group(P<0.05).Neither group showed significant adverse reactions during the treatment period.Conclusion:Fu Zheng Bu Tu acupuncture-moxibustion plus dynamic-static balance rehabilitation training can effectively alleviate muscle spasticity,improve balance ability,enhance sEMG signal levels,modulate hemodynamics,and promote the recovery of limb motor function in HAS patients.
