目的探讨细胞信号传导分子SMAD2和血清和糖皮质激素诱导的蛋白激酶3(Serum and glucocorticoid-inducible protein kinase 3,SGK3)在非小细胞肺癌(Non small cell lung cancer,NSCLC)中的表达及临床意义。方法选取郑州市第六人民医院和...目的探讨细胞信号传导分子SMAD2和血清和糖皮质激素诱导的蛋白激酶3(Serum and glucocorticoid-inducible protein kinase 3,SGK3)在非小细胞肺癌(Non small cell lung cancer,NSCLC)中的表达及临床意义。方法选取郑州市第六人民医院和安阳市肿瘤医院2010年6月至2013年6月收治的98例NSCLC患者作为研究对象,采用系统回顾性分析法分析所有患者的临床资料,根据其资料结果收集98例NSCLC患者其98份癌组织标本(研究组)及98份癌旁组织标本(对照组)作为研究对象,所有组织标本均行SMAD2与SGK3蛋白的检测,并分析二者的表达与NSCLC患者临床病理特征的关系。结果①NSCLC组织中SMAD2与SGK3蛋白阳性表达率分别为77.55%和74.49%,明显高于癌旁组织中SMAD2与SGK3蛋白阳性表达(17.35%与15.31%)(P<0.05);②SMAD2蛋白表达与NSCLC患者性别、年龄、病理类型、肿瘤大小均无明显关系(P>0.05);SMAD2蛋白表达与NSCLC患者肿瘤分化程度、淋巴结转移存在显著相关性(P<0.05);③SGK3蛋白表达与NSCLC患者性别、年龄、病理类型、肿瘤大小均无明显关系(P>0.05);SGK3蛋白表达与NSCLC患者肿瘤分化程度、淋巴结转移存在显著相关性(P<0.05);④经单因素分析得出:低分化、淋巴结转移、SMAD2蛋白阳性表达、SGK3蛋白阳性表达患者生存时间均显著缩短(P<0.05);⑤经多因素Cox分析得出:低分化、有淋巴结转移、SMAD2蛋白阳性、SGK3蛋白阳性均为影响NSCLC患者预后的独立危险因素(P<0.05)。结论SMAD2与SGK3蛋白在NSCLC组织中表达显著高于癌旁组织,其表达水平与NSCLC患者分化程度、淋巴结转移均有显著相关,可能参与了NSCLC的发生、发展。展开更多
Objective:Myeloma bone disease(MBD)is the most common complication of multiple myeloma(MM).Our previous study showed that the serum levels of C3/C4 in MM patients were significantly positively correlated with the seve...Objective:Myeloma bone disease(MBD)is the most common complication of multiple myeloma(MM).Our previous study showed that the serum levels of C3/C4 in MM patients were significantly positively correlated with the severity of bone disease.However,the mechanism of C3 a/C4 a in osteoclasts MM patients remains unclear.Methods:The formation and function of osteoclasts were analyzed after adding C3 a/C4 a in vitro.RNA-seq analysis was used to screen the potential pathways affecting osteoclasts,and the results were verified by Western blot,q RT-PCR,and pathway inhibitors.Results:The osteoclast area per view induced by 1μg/m L(mean±SD:50.828±12.984%)and 10μg/m L(53.663±12.685%)of C3 a was significantly increased compared to the control group(0μg/m L)(34.635±8.916%)(P<0.001 and P<0.001,respectively).The relative m RNA expressions of genes,OSCAR/TRAP/RANKL/cathepsin K,induced by 1μg/m L(median:5.041,3.726,1.638,and 4.752,respectively)and 10μg/m L(median:5.140,3.702,2.250,and 5.172,respectively)of C3 a was significantly increased compared to the control group(median:3.137,2.004,0.573,and 2.257,respectively)(1μg/m L P=0.001,P=0.003,P<0.001,and P=0.008,respectively;10μg/m L:P<0.001,P=0.019,P<0.001,and P=0.002,respectively).The absorption areas of the osteoclast resorption pits per view induced by 1μg/m L(mean±SD:51.464±11.983%)and 10μg/m L(50.219±12.067%)of C3 a was also significantly increased(33.845±8.331%)(P<0.001 and P<0.001,respectively)compared to the control.There was no difference between the C4 a and control groups.RNA-seq analysis showed that C3 a promoted the proliferation of osteoclasts using the phosphoinositide 3-kinase(PI3 K)signaling pathway.The relative expressions of PIK3 CA/phosphoinositide dependent kinase-1(PDK1)/serum and glucocorticoid inducible protein kinases(SGK3)genes and PI3 K/PDK1/p-SGK3 protein in the C3 a group were significantly higher than in the control group.The activation role of C3 a in osteoclasts of MM patients was reduced by the SGK inhibitor(EMD638683).Conclusions:C3 a activated osteoclasts by regulating the PI3 K/PDK1/SGK3 pathways in MM patients,which was reduced using a SGK inhibitor.Overall,our results identified potential therapeutic targets and strategies for MBD patients。展开更多
目的血清和糖皮质激素诱导的蛋白激酶3(serum and glucocorticoid-inducible protein kinase 3,SGK3)参与细胞增殖、分化和物质转运等生命活动,在某些肿瘤形成中担任重要角色。本研究设计观察SGK3过表达对乳腺癌细胞株MDA-MB-231细胞增...目的血清和糖皮质激素诱导的蛋白激酶3(serum and glucocorticoid-inducible protein kinase 3,SGK3)参与细胞增殖、分化和物质转运等生命活动,在某些肿瘤形成中担任重要角色。本研究设计观察SGK3过表达对乳腺癌细胞株MDA-MB-231细胞增殖和存活的影响,并初步探讨其作用的分子机制。方法免疫组化方法检测乳腺癌组织中SGK3蛋白的表达;通过GenEscort TMⅠ介导用重组质粒pEGFPN1-SGK3瞬时转染乳腺癌MDA-MB-231细胞,荧光显微镜和蛋白质印迹法鉴定转染细胞中融合蛋白SGK3-GFP的表达;细胞增殖实验观察转染细胞的生长增殖情况;流式细胞术分析转染细胞的细胞周期时相变化;蛋白质印迹方法检测细胞增殖与存活相关基因的表达。结果 SGK3在乳腺癌组织中的表达量明显高于癌旁组织和正常乳腺组织(F=29.672,P<0.001),与二者组间比较,均P<0.01。荧光显微镜下可见转染细胞因外源基因的表达而发出绿色荧光,且蛋白质印迹法检测可见融合蛋白SGK3-GFP的表达;过表达SGK3的MDA-MB-231细胞,其细胞生长速度加快,细胞倍增时间缩短(F=19.557,P=0.001),与亲本细胞及转染空载体细胞组间比较均P<0.01。与亲本细胞和转染空载体组比较,SGK3过表达使细胞凋亡比例明显下降(F=12.156,P=0.008),与前两者组间比较P值分别为0.019和0.030;细胞内源性Bad的表达量降低(F=11.152,P=0.010),与前两者组间比较P值分别为0.008和0.005;而Bcl-xL(F=8.810,P=0.016,与前两者组间比较P值分别为0.014和0.009)以及p-GSK3β(F=42.253,P<0.001,与前两者组间比较P值分别为<0.001和0.009)的表达量均升高。结论乳腺癌组织高表达SGK3,SGK3过表达可促进MDA-MB-231细胞的增殖与存活,影响细胞增殖与存活相关基因的表达可能是其发挥作用的主要分子机制。展开更多
目的通过浸润性乳腺癌中血清和糖皮质激素调节蛋白激酶3(serum and glucocorticoid-regulated protein kinase 3,SGK3)的表达情况进行分析,初步探讨SGK3与浸润性乳腺癌临床病理特征的相关性。方法课题组于2015年10月—2016年1月,利用组...目的通过浸润性乳腺癌中血清和糖皮质激素调节蛋白激酶3(serum and glucocorticoid-regulated protein kinase 3,SGK3)的表达情况进行分析,初步探讨SGK3与浸润性乳腺癌临床病理特征的相关性。方法课题组于2015年10月—2016年1月,利用组织芯片,检测SGK3在癌旁乳腺组织和浸润性乳腺癌组织中的表达,并结合临床参数探讨SGK3与浸润性乳腺癌临床病理特征的关系,计量资料比较采用t检验,P<0.05为差异有统计学意义。结果免疫组织化学(组化)结果显示,SGK3在浸润性乳腺癌中的表达高于癌旁正常乳腺组织[(338.157±133.349)、(199.959±52.917)],对比差异有统计学意义(P<0.05),且在浸润性乳腺癌组织中,雌激素受体(estrogen receptor,ER)阳性组织中SGK3的表达较ER阴性组织增高[(378.494±142.749)、(318.853±143.199)],对比差异有统计学意义(P<0.05)。结论 SGK3在浸润性乳腺癌组织中呈高表达,且在ER阳性的浸润性乳腺癌组织中的表达高于ER阴性组织,SGK3有可能成为浸润性乳腺癌临床诊断和预后的指标。展开更多
文摘目的探讨细胞信号传导分子SMAD2和血清和糖皮质激素诱导的蛋白激酶3(Serum and glucocorticoid-inducible protein kinase 3,SGK3)在非小细胞肺癌(Non small cell lung cancer,NSCLC)中的表达及临床意义。方法选取郑州市第六人民医院和安阳市肿瘤医院2010年6月至2013年6月收治的98例NSCLC患者作为研究对象,采用系统回顾性分析法分析所有患者的临床资料,根据其资料结果收集98例NSCLC患者其98份癌组织标本(研究组)及98份癌旁组织标本(对照组)作为研究对象,所有组织标本均行SMAD2与SGK3蛋白的检测,并分析二者的表达与NSCLC患者临床病理特征的关系。结果①NSCLC组织中SMAD2与SGK3蛋白阳性表达率分别为77.55%和74.49%,明显高于癌旁组织中SMAD2与SGK3蛋白阳性表达(17.35%与15.31%)(P<0.05);②SMAD2蛋白表达与NSCLC患者性别、年龄、病理类型、肿瘤大小均无明显关系(P>0.05);SMAD2蛋白表达与NSCLC患者肿瘤分化程度、淋巴结转移存在显著相关性(P<0.05);③SGK3蛋白表达与NSCLC患者性别、年龄、病理类型、肿瘤大小均无明显关系(P>0.05);SGK3蛋白表达与NSCLC患者肿瘤分化程度、淋巴结转移存在显著相关性(P<0.05);④经单因素分析得出:低分化、淋巴结转移、SMAD2蛋白阳性表达、SGK3蛋白阳性表达患者生存时间均显著缩短(P<0.05);⑤经多因素Cox分析得出:低分化、有淋巴结转移、SMAD2蛋白阳性、SGK3蛋白阳性均为影响NSCLC患者预后的独立危险因素(P<0.05)。结论SMAD2与SGK3蛋白在NSCLC组织中表达显著高于癌旁组织,其表达水平与NSCLC患者分化程度、淋巴结转移均有显著相关,可能参与了NSCLC的发生、发展。
基金supported by the National Natural Science Foundation of China(Grant Nos.81770110,81900131,and 82000219)the Anticancer Major Special Project of Tianjin(Grant No.12ZCDZSY18000)+4 种基金the Tianjin Municipal Natural Science Foundation(Grant Nos.18JCYBJC27200 and 18JCQNJC80400)the Tianjin Education Commission Research Project(Grant Nos.2018KJ043 and 2018KJ045)the Tianjin Health and Family Planning Commission(Grant No.15KG150)the Youth Incubation Fund of Tianjin Medical University General Hospital(Grant No.ZYYFY2019020)the Tianjin Science and Technology Planning Project(Grant No.20YFZCSY00060)。
文摘Objective:Myeloma bone disease(MBD)is the most common complication of multiple myeloma(MM).Our previous study showed that the serum levels of C3/C4 in MM patients were significantly positively correlated with the severity of bone disease.However,the mechanism of C3 a/C4 a in osteoclasts MM patients remains unclear.Methods:The formation and function of osteoclasts were analyzed after adding C3 a/C4 a in vitro.RNA-seq analysis was used to screen the potential pathways affecting osteoclasts,and the results were verified by Western blot,q RT-PCR,and pathway inhibitors.Results:The osteoclast area per view induced by 1μg/m L(mean±SD:50.828±12.984%)and 10μg/m L(53.663±12.685%)of C3 a was significantly increased compared to the control group(0μg/m L)(34.635±8.916%)(P<0.001 and P<0.001,respectively).The relative m RNA expressions of genes,OSCAR/TRAP/RANKL/cathepsin K,induced by 1μg/m L(median:5.041,3.726,1.638,and 4.752,respectively)and 10μg/m L(median:5.140,3.702,2.250,and 5.172,respectively)of C3 a was significantly increased compared to the control group(median:3.137,2.004,0.573,and 2.257,respectively)(1μg/m L P=0.001,P=0.003,P<0.001,and P=0.008,respectively;10μg/m L:P<0.001,P=0.019,P<0.001,and P=0.002,respectively).The absorption areas of the osteoclast resorption pits per view induced by 1μg/m L(mean±SD:51.464±11.983%)and 10μg/m L(50.219±12.067%)of C3 a was also significantly increased(33.845±8.331%)(P<0.001 and P<0.001,respectively)compared to the control.There was no difference between the C4 a and control groups.RNA-seq analysis showed that C3 a promoted the proliferation of osteoclasts using the phosphoinositide 3-kinase(PI3 K)signaling pathway.The relative expressions of PIK3 CA/phosphoinositide dependent kinase-1(PDK1)/serum and glucocorticoid inducible protein kinases(SGK3)genes and PI3 K/PDK1/p-SGK3 protein in the C3 a group were significantly higher than in the control group.The activation role of C3 a in osteoclasts of MM patients was reduced by the SGK inhibitor(EMD638683).Conclusions:C3 a activated osteoclasts by regulating the PI3 K/PDK1/SGK3 pathways in MM patients,which was reduced using a SGK inhibitor.Overall,our results identified potential therapeutic targets and strategies for MBD patients。
文摘目的血清和糖皮质激素诱导的蛋白激酶3(serum and glucocorticoid-inducible protein kinase 3,SGK3)参与细胞增殖、分化和物质转运等生命活动,在某些肿瘤形成中担任重要角色。本研究设计观察SGK3过表达对乳腺癌细胞株MDA-MB-231细胞增殖和存活的影响,并初步探讨其作用的分子机制。方法免疫组化方法检测乳腺癌组织中SGK3蛋白的表达;通过GenEscort TMⅠ介导用重组质粒pEGFPN1-SGK3瞬时转染乳腺癌MDA-MB-231细胞,荧光显微镜和蛋白质印迹法鉴定转染细胞中融合蛋白SGK3-GFP的表达;细胞增殖实验观察转染细胞的生长增殖情况;流式细胞术分析转染细胞的细胞周期时相变化;蛋白质印迹方法检测细胞增殖与存活相关基因的表达。结果 SGK3在乳腺癌组织中的表达量明显高于癌旁组织和正常乳腺组织(F=29.672,P<0.001),与二者组间比较,均P<0.01。荧光显微镜下可见转染细胞因外源基因的表达而发出绿色荧光,且蛋白质印迹法检测可见融合蛋白SGK3-GFP的表达;过表达SGK3的MDA-MB-231细胞,其细胞生长速度加快,细胞倍增时间缩短(F=19.557,P=0.001),与亲本细胞及转染空载体细胞组间比较均P<0.01。与亲本细胞和转染空载体组比较,SGK3过表达使细胞凋亡比例明显下降(F=12.156,P=0.008),与前两者组间比较P值分别为0.019和0.030;细胞内源性Bad的表达量降低(F=11.152,P=0.010),与前两者组间比较P值分别为0.008和0.005;而Bcl-xL(F=8.810,P=0.016,与前两者组间比较P值分别为0.014和0.009)以及p-GSK3β(F=42.253,P<0.001,与前两者组间比较P值分别为<0.001和0.009)的表达量均升高。结论乳腺癌组织高表达SGK3,SGK3过表达可促进MDA-MB-231细胞的增殖与存活,影响细胞增殖与存活相关基因的表达可能是其发挥作用的主要分子机制。
文摘目的通过浸润性乳腺癌中血清和糖皮质激素调节蛋白激酶3(serum and glucocorticoid-regulated protein kinase 3,SGK3)的表达情况进行分析,初步探讨SGK3与浸润性乳腺癌临床病理特征的相关性。方法课题组于2015年10月—2016年1月,利用组织芯片,检测SGK3在癌旁乳腺组织和浸润性乳腺癌组织中的表达,并结合临床参数探讨SGK3与浸润性乳腺癌临床病理特征的关系,计量资料比较采用t检验,P<0.05为差异有统计学意义。结果免疫组织化学(组化)结果显示,SGK3在浸润性乳腺癌中的表达高于癌旁正常乳腺组织[(338.157±133.349)、(199.959±52.917)],对比差异有统计学意义(P<0.05),且在浸润性乳腺癌组织中,雌激素受体(estrogen receptor,ER)阳性组织中SGK3的表达较ER阴性组织增高[(378.494±142.749)、(318.853±143.199)],对比差异有统计学意义(P<0.05)。结论 SGK3在浸润性乳腺癌组织中呈高表达,且在ER阳性的浸润性乳腺癌组织中的表达高于ER阴性组织,SGK3有可能成为浸润性乳腺癌临床诊断和预后的指标。
