浙江医学
浙江醫學
절강의학
ZHEJIANG MEDICAL JOURNAL
2014年
9期
742-746
,共5页
细梗香草%肺癌%凋亡%活性氧
細梗香草%肺癌%凋亡%活性氧
세경향초%폐암%조망%활성양
Capilliposide%Lung cancer%Apoptosis%ROS
目的：观察细梗香草总皂苷（LC）对非小细胞肺癌H460细胞增殖和凋亡的影响。方法在H460肺癌细胞培养时加入不同浓度的LC，采用MTT法和克隆形成实验检测细胞的增殖活性，AnnexinⅤ/PI（FCM）法观察LC对肺癌细胞凋亡的诱导作用和周期分布的影响。DCFH- DA荧光探针FCM检测细胞内活性氧生成，Western blot法检测NF-κB、I-κB、Bax、Bcl-2、Capase-3的蛋白表达。结果 LC可抑制H460细胞生长，具有剂量、时间依赖性；LC作用H460细胞后克隆形成率明显下降。LC可引起H460细胞凋亡，并可将H460生长阻滞在S期。LC以浓度依赖方式增高H460细胞内活性氧水平，还可使H460细胞I-κB、Bax和Capase-3表达增加，而使NF-κB和Bcl-2表达降低。结论 LC可以抑制H460细胞增殖，可能是通过增加细胞内活性氧表达，激活了肺癌细胞线粒体凋亡途径。
目的：觀察細梗香草總皂苷（LC）對非小細胞肺癌H460細胞增殖和凋亡的影響。方法在H460肺癌細胞培養時加入不同濃度的LC，採用MTT法和剋隆形成實驗檢測細胞的增殖活性，AnnexinⅤ/PI（FCM）法觀察LC對肺癌細胞凋亡的誘導作用和週期分佈的影響。DCFH- DA熒光探針FCM檢測細胞內活性氧生成，Western blot法檢測NF-κB、I-κB、Bax、Bcl-2、Capase-3的蛋白錶達。結果 LC可抑製H460細胞生長，具有劑量、時間依賴性；LC作用H460細胞後剋隆形成率明顯下降。LC可引起H460細胞凋亡，併可將H460生長阻滯在S期。LC以濃度依賴方式增高H460細胞內活性氧水平，還可使H460細胞I-κB、Bax和Capase-3錶達增加，而使NF-κB和Bcl-2錶達降低。結論 LC可以抑製H460細胞增殖，可能是通過增加細胞內活性氧錶達，激活瞭肺癌細胞線粒體凋亡途徑。
목적：관찰세경향초총조감（LC）대비소세포폐암H460세포증식화조망적영향。방법재H460폐암세포배양시가입불동농도적LC，채용MTT법화극륭형성실험검측세포적증식활성，AnnexinⅤ/PI（FCM）법관찰LC대폐암세포조망적유도작용화주기분포적영향。DCFH- DA형광탐침FCM검측세포내활성양생성，Western blot법검측NF-κB、I-κB、Bax、Bcl-2、Capase-3적단백표체。결과 LC가억제H460세포생장，구유제량、시간의뢰성；LC작용H460세포후극륭형성솔명현하강。LC가인기H460세포조망，병가장H460생장조체재S기。LC이농도의뢰방식증고H460세포내활성양수평，환가사H460세포I-κB、Bax화Capase-3표체증가，이사NF-κB화Bcl-2표체강저。결론 LC가이억제H460세포증식，가능시통과증가세포내활성양표체，격활료폐암세포선립체조망도경。
Objective To investigate the effect of capil iposides isolated from Lysimachia capil ipes Hemsl(capil ipes, LC) on proliferation and apoptosis of human non- smal lung cancer cellline H460 in vitro. Methods H460 cells were cultured with different concentration of LC. The proliferation, colony formation and cellcycle of H460 cells were measured by MTT method, colony formation assay and flow cytometry, respectively. Intracellular reactive oxygen species (ROS) was measured by FCM with fluorescent probe of DCFH- DA. The expression of signal transduction proteins NF- κB, I- κB, Bax, Bcl- 2 and capase- 3 was detected by Western blotting. Results The proliferation of H460 cells was inhibited and the ability of colony- formation was re-duced;the apoptosis rate of H460 cells was increased(P<0.05)and the cellcycle was arrested at S phase(P<0.05)after cul-tured with LC for 24h. LC significantly increased ROS level in H460 cells in a concentration- dependent manner. LC up- regulated I- κB and Bax expression,and activated capase- 3 enzyme (P<0.05)while the expression levels of NF- ΚB and Bcl- 2 were down- regulated(P<0.05). Conclusion LC can inhibit the growth and induce apoptosis of H460 cells, which may be associated with ROS generation and activation of mitochondrial apoptotic pathway.
