目的:探讨体外沉默Kruppel样因子4(Kruppel like factor 4,KLF4)基因的表达对食管癌KYSE140细胞增殖及迁移的影响。方法:Western blotting法检测人食管癌细胞株KYSE140、KYSE150、EC109及EC9706及食管永生化细胞NE3中KLF4蛋白的表达,化...目的:探讨体外沉默Kruppel样因子4(Kruppel like factor 4,KLF4)基因的表达对食管癌KYSE140细胞增殖及迁移的影响。方法:Western blotting法检测人食管癌细胞株KYSE140、KYSE150、EC109及EC9706及食管永生化细胞NE3中KLF4蛋白的表达,化学合成2对靶向KLF4的siRNA(KLF4-siRNA1,KLF4-siRNA2),并设对照siRNA(Ctrl-siRNA),分别体外转染至高表达KLF4的食管癌KYSE140细胞中,形成KLF4-siRNA1-KYSE140、KLF4-siRNA2-KYSE140及Ctrl-siRNA-KYSE140细胞,通过MTT实验、Transwell实验分别检测转染后食管癌KYSE140细胞的增殖及迁移。结果:食管癌细胞株KYSE140中KLF4蛋白的表达明显高于KYSE150、EC109及EC9706细胞株[(5.62±0.02)vs(1.71±0.23)、(3.24±0.35)、(3.16±0.41),均P<0.05]。KLF4-siRNA1-KYSE140、KLF4-siRNA2-KYSE140与Ctrl-siRNA-KYSE140细胞相比,KLF4蛋白表达明显降低[(0.49±0.18)、(0.32±0.09)vs(0.98±0.19),均P<0.05],细胞增殖能力明显增高[(1.2±0.8)、(1.4±0.1)vs(0.6±0.1),均P<0.05],迁移细胞数量也明显增加[(780±22)、(475±25)vs(83±17)个,P<0.05]。结论:KLF4在人食管癌细胞的增殖和迁移过程中起着负调控作用。展开更多
目的观察川芎嗪联合顺铂对Lewis肺癌小鼠移植瘤生长及微血管生成的影响。方法将Lewis肺癌细胞接种于C57BL/6小鼠右腋皮下,按数字表法随机分为4组:模型组(0.9%氯化钠,0.2 m L),川芎嗪组(100 mg·kg^(-1),0.2m L),顺铂组(2 mg·kg...目的观察川芎嗪联合顺铂对Lewis肺癌小鼠移植瘤生长及微血管生成的影响。方法将Lewis肺癌细胞接种于C57BL/6小鼠右腋皮下,按数字表法随机分为4组:模型组(0.9%氯化钠,0.2 m L),川芎嗪组(100 mg·kg^(-1),0.2m L),顺铂组(2 mg·kg^(-1),0.2 m L),联合组(如上述同等剂量,0.2 m L),每组14只。于接种后第7天起,分别连续给药14 d,监测皮下移植瘤体积变化。于接种后第21天,处死全部小鼠,剥离皮下肿瘤,称重并计算抑瘤率以及肿瘤坏死情况,用蛋白印迹方法检测锌指蛋白4(KLF4)与血管内皮生长因子(VEGF)表达,微血管密度(MVD)计数由CD105染色确定。结果各用药组,肿瘤的生长明显受到抑制,瘤重明显低于模型组(P<0.05);联合治疗组的抑瘤率明显高于川芎嗪组与顺铂组(62.48%vs 35.58%vs 40.41%,P<0.01)。联合治疗组,肿瘤坏死率明显高于顺铂组[(31.76±15.20)%vs(24.64±11.96)%,P<0.05]。与模型组比较,各用药组的肿瘤组织中VEGF表达与MVD计数水平均显著降低(均P<0.05);KLF4的表达水平显著增加(均P<0.05));而川芎嗪组与顺铂组比较,无明显差异(P>0.05)。结论川芎嗪能够抑制Lewis肺癌的生长和微血管生成,与顺铂联合具有协同作用,这可能与其下调VEGF的表达与上调KLF4的表达有关。展开更多
Objective Hepatic stellate cells(HSCs)play a crucial role in liver fibrosis.Early-stage liver fibrosis is reversible and intimately associated with the state of HSCs.Kruppel-like factor 4(KLF4)plays a pivotal role in ...Objective Hepatic stellate cells(HSCs)play a crucial role in liver fibrosis.Early-stage liver fibrosis is reversible and intimately associated with the state of HSCs.Kruppel-like factor 4(KLF4)plays a pivotal role in a wide array of physiological and pathological processes.This study aimed to investigate the effect of KLF4 on the proliferation,apoptosis and phenotype of quiescent HSCs Methods We designed a KLF4 lentiviral vector and a KLF4 siRNA lentiviral vector,to upregulate and silence KLF4 expression in human HSC LX-2 cells via transfection.Cell proliferation was assessed using the CCK-8 assay.Flow cytometry was used to detect the cell cycle distribution and apoptosis rate.Western blotting was used to determine the levels of some quiescence and activation markers of HSCs Results Overexpression of KLF4 significantly increased the levels of E-cadherin and ZO-1,which are quiescent HSC markers,while significantly decreased the levels of N-cadherin and a-SMA,known activated HSC markers.In contrast,cell proliferation and apoptosis rates were elevated in LX-2 cells in which KLF4 expression was silenced Conclusion KLF4 inhibits the proliferation and activation of human LX-2 HSCs.It might be a key regulatory protein in the maintenance of HSC quiescence and may serve as a target for the inhibition of hepatic fibrosis.展开更多
文摘目的:探讨体外沉默Kruppel样因子4(Kruppel like factor 4,KLF4)基因的表达对食管癌KYSE140细胞增殖及迁移的影响。方法:Western blotting法检测人食管癌细胞株KYSE140、KYSE150、EC109及EC9706及食管永生化细胞NE3中KLF4蛋白的表达,化学合成2对靶向KLF4的siRNA(KLF4-siRNA1,KLF4-siRNA2),并设对照siRNA(Ctrl-siRNA),分别体外转染至高表达KLF4的食管癌KYSE140细胞中,形成KLF4-siRNA1-KYSE140、KLF4-siRNA2-KYSE140及Ctrl-siRNA-KYSE140细胞,通过MTT实验、Transwell实验分别检测转染后食管癌KYSE140细胞的增殖及迁移。结果:食管癌细胞株KYSE140中KLF4蛋白的表达明显高于KYSE150、EC109及EC9706细胞株[(5.62±0.02)vs(1.71±0.23)、(3.24±0.35)、(3.16±0.41),均P<0.05]。KLF4-siRNA1-KYSE140、KLF4-siRNA2-KYSE140与Ctrl-siRNA-KYSE140细胞相比,KLF4蛋白表达明显降低[(0.49±0.18)、(0.32±0.09)vs(0.98±0.19),均P<0.05],细胞增殖能力明显增高[(1.2±0.8)、(1.4±0.1)vs(0.6±0.1),均P<0.05],迁移细胞数量也明显增加[(780±22)、(475±25)vs(83±17)个,P<0.05]。结论:KLF4在人食管癌细胞的增殖和迁移过程中起着负调控作用。
文摘目的观察川芎嗪联合顺铂对Lewis肺癌小鼠移植瘤生长及微血管生成的影响。方法将Lewis肺癌细胞接种于C57BL/6小鼠右腋皮下,按数字表法随机分为4组:模型组(0.9%氯化钠,0.2 m L),川芎嗪组(100 mg·kg^(-1),0.2m L),顺铂组(2 mg·kg^(-1),0.2 m L),联合组(如上述同等剂量,0.2 m L),每组14只。于接种后第7天起,分别连续给药14 d,监测皮下移植瘤体积变化。于接种后第21天,处死全部小鼠,剥离皮下肿瘤,称重并计算抑瘤率以及肿瘤坏死情况,用蛋白印迹方法检测锌指蛋白4(KLF4)与血管内皮生长因子(VEGF)表达,微血管密度(MVD)计数由CD105染色确定。结果各用药组,肿瘤的生长明显受到抑制,瘤重明显低于模型组(P<0.05);联合治疗组的抑瘤率明显高于川芎嗪组与顺铂组(62.48%vs 35.58%vs 40.41%,P<0.01)。联合治疗组,肿瘤坏死率明显高于顺铂组[(31.76±15.20)%vs(24.64±11.96)%,P<0.05]。与模型组比较,各用药组的肿瘤组织中VEGF表达与MVD计数水平均显著降低(均P<0.05);KLF4的表达水平显著增加(均P<0.05));而川芎嗪组与顺铂组比较,无明显差异(P>0.05)。结论川芎嗪能够抑制Lewis肺癌的生长和微血管生成,与顺铂联合具有协同作用,这可能与其下调VEGF的表达与上调KLF4的表达有关。
基金supported by the National Natural Science Foundation of China(No.81071541).
文摘Objective Hepatic stellate cells(HSCs)play a crucial role in liver fibrosis.Early-stage liver fibrosis is reversible and intimately associated with the state of HSCs.Kruppel-like factor 4(KLF4)plays a pivotal role in a wide array of physiological and pathological processes.This study aimed to investigate the effect of KLF4 on the proliferation,apoptosis and phenotype of quiescent HSCs Methods We designed a KLF4 lentiviral vector and a KLF4 siRNA lentiviral vector,to upregulate and silence KLF4 expression in human HSC LX-2 cells via transfection.Cell proliferation was assessed using the CCK-8 assay.Flow cytometry was used to detect the cell cycle distribution and apoptosis rate.Western blotting was used to determine the levels of some quiescence and activation markers of HSCs Results Overexpression of KLF4 significantly increased the levels of E-cadherin and ZO-1,which are quiescent HSC markers,while significantly decreased the levels of N-cadherin and a-SMA,known activated HSC markers.In contrast,cell proliferation and apoptosis rates were elevated in LX-2 cells in which KLF4 expression was silenced Conclusion KLF4 inhibits the proliferation and activation of human LX-2 HSCs.It might be a key regulatory protein in the maintenance of HSC quiescence and may serve as a target for the inhibition of hepatic fibrosis.
