目的探讨CLCA4对结直肠癌细胞增殖、侵袭、迁移及上皮-间质转化的影响及可能机制。方法收集100例患者的结直肠癌组织及癌旁正常组织。培养SW620细胞,根据转染质粒不同分为Vector组(转染pcDNA3.1)、CLCA4组(转染pcDNA3.1-CLCA4)、sh-NC组...目的探讨CLCA4对结直肠癌细胞增殖、侵袭、迁移及上皮-间质转化的影响及可能机制。方法收集100例患者的结直肠癌组织及癌旁正常组织。培养SW620细胞,根据转染质粒不同分为Vector组(转染pcDNA3.1)、CLCA4组(转染pcDNA3.1-CLCA4)、sh-NC组(转染sh-NC)、sh-CLCA4组(转染sh-CLCA4)及Control组(无处理)。sh-NC组及sh-CLCA4组细胞用GSK2126458分处理并定义为sh-NC+GSK2126458组、sh-CLCA4+GSK2126458组。分析CLCA4与结直肠癌临床病理特征、预后的关系,CCK-8检测细胞增殖能力,Tanswell实验检测细胞侵袭能力,划痕实验检测细胞迁移能力,qRT-PCR检测细胞、组织中CLCA4 mRNA表达水平,Western blotting检测细胞或组织中CLCA4、E-cadherin、N-cadherin、Vimentin、PI3K、p-PI3K、AKT、p-AKT、mTOR、p-mTOR蛋白表达水平。结果结直肠癌组织CLCA4 mRNA、蛋白表达水平低于癌旁正常组织(P<0.05)。CLCA4表达水平与临床分期、淋巴结转移、脉管侵犯、神经侵犯及血清癌胚抗原水平有关(P<0.05)。CLCA4高表达组患者总体生存率高于CLCA4低表达组(P=0.006)。CLCA4组细胞48 h、72 h OD值、侵袭细胞数,Vimentin、N-cadherin表达水平及p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR值低于Vector组,迁移率及CLCA4、E-cadherin表达水平高于Vector组(P<0.05);sh-CLCA4组细胞48 h、72 h OD值、侵袭细胞数,Vimentin、N-cadherin表达水平及p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR值高于sh-NC组,迁移率及CLCA4、E-cadherin表达水平低于sh-NC组(P<0.05);sh-NC+GSK2126458组细胞48 h、72 h OD值、侵袭细胞数低于sh-NC组,迁移率高于sh-NC组(P<0.05),sh-CLCA4+GSK2126458组细胞48 h、72 h OD值、侵袭细胞数低于sh-CLCA4组,迁移率高于sh-CLCA4组(P<0.05)。结论CLCA4在结直肠癌组织中低表达,与肿瘤临床病理特征、预后有关,CLCA4通过PI3K/AKT/mTOR信号通路抑制结直肠癌细胞的增殖、侵袭、迁移及上皮-间质转化。展开更多
Objective A calcium-activated chloride current （ICl（Ca）） has been observed in medium-sized sensory neurons of the dorsal root ganglion （DRG）. Axotomy of the sciatic nerve induces a similar current in the majorit...Objective A calcium-activated chloride current （ICl（Ca）） has been observed in medium-sized sensory neurons of the dorsal root ganglion （DRG）. Axotomy of the sciatic nerve induces a similar current in the majority of medium and large diameter neurons. Our aim is to identify the molecule（s） underlying this current. Methods Using conventional and quantitative RT-PCR, we examined the expression in DRG of members of three families of genes, which have been shown to have latch） current inducing properties. Results We showed the detection of transcripts representing several members of these families, i.e. chloride channel calciumactivated （CLCA）, Bestrophin and Tweety gene families in adult DRG, in the normal state and 3 d after sciatic nerve section, a model for peripheral nerve injury. Conclusion Our analysis revealed that that mBestl and Tweety2 appear as the best candidates to play a role in the injury-induced Icl（Ca） in DRG neurons.展开更多
文摘目的探讨CLCA4对结直肠癌细胞增殖、侵袭、迁移及上皮-间质转化的影响及可能机制。方法收集100例患者的结直肠癌组织及癌旁正常组织。培养SW620细胞,根据转染质粒不同分为Vector组(转染pcDNA3.1)、CLCA4组(转染pcDNA3.1-CLCA4)、sh-NC组(转染sh-NC)、sh-CLCA4组(转染sh-CLCA4)及Control组(无处理)。sh-NC组及sh-CLCA4组细胞用GSK2126458分处理并定义为sh-NC+GSK2126458组、sh-CLCA4+GSK2126458组。分析CLCA4与结直肠癌临床病理特征、预后的关系,CCK-8检测细胞增殖能力,Tanswell实验检测细胞侵袭能力,划痕实验检测细胞迁移能力,qRT-PCR检测细胞、组织中CLCA4 mRNA表达水平,Western blotting检测细胞或组织中CLCA4、E-cadherin、N-cadherin、Vimentin、PI3K、p-PI3K、AKT、p-AKT、mTOR、p-mTOR蛋白表达水平。结果结直肠癌组织CLCA4 mRNA、蛋白表达水平低于癌旁正常组织(P<0.05)。CLCA4表达水平与临床分期、淋巴结转移、脉管侵犯、神经侵犯及血清癌胚抗原水平有关(P<0.05)。CLCA4高表达组患者总体生存率高于CLCA4低表达组(P=0.006)。CLCA4组细胞48 h、72 h OD值、侵袭细胞数,Vimentin、N-cadherin表达水平及p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR值低于Vector组,迁移率及CLCA4、E-cadherin表达水平高于Vector组(P<0.05);sh-CLCA4组细胞48 h、72 h OD值、侵袭细胞数,Vimentin、N-cadherin表达水平及p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR值高于sh-NC组,迁移率及CLCA4、E-cadherin表达水平低于sh-NC组(P<0.05);sh-NC+GSK2126458组细胞48 h、72 h OD值、侵袭细胞数低于sh-NC组,迁移率高于sh-NC组(P<0.05),sh-CLCA4+GSK2126458组细胞48 h、72 h OD值、侵袭细胞数低于sh-CLCA4组,迁移率高于sh-CLCA4组(P<0.05)。结论CLCA4在结直肠癌组织中低表达,与肿瘤临床病理特征、预后有关,CLCA4通过PI3K/AKT/mTOR信号通路抑制结直肠癌细胞的增殖、侵袭、迁移及上皮-间质转化。
文摘Objective A calcium-activated chloride current （ICl（Ca）） has been observed in medium-sized sensory neurons of the dorsal root ganglion （DRG）. Axotomy of the sciatic nerve induces a similar current in the majority of medium and large diameter neurons. Our aim is to identify the molecule（s） underlying this current. Methods Using conventional and quantitative RT-PCR, we examined the expression in DRG of members of three families of genes, which have been shown to have latch） current inducing properties. Results We showed the detection of transcripts representing several members of these families, i.e. chloride channel calciumactivated （CLCA）, Bestrophin and Tweety gene families in adult DRG, in the normal state and 3 d after sciatic nerve section, a model for peripheral nerve injury. Conclusion Our analysis revealed that that mBestl and Tweety2 appear as the best candidates to play a role in the injury-induced Icl（Ca） in DRG neurons.
