摘要
目的:观察芍药苷诱导非小细胞肺癌细胞凋亡的作用,并探讨其作用机制。方法:采用细胞增殖与活性检测法(CCK-8)检测不同浓度芍药苷(2.5、5、10、20、25μmol·L-1)对H1299、H292、A549细胞增殖的抑制率,筛选合适药物浓度及实验细胞。克隆形成实验检测芍药苷对肺癌细胞抑制作用。采用膜联蛋白V-异硫氰酸荧光素(AnnexinV-FITC)/碘化丙锭(PI)染色,流式细胞仪检测芍药苷对细胞凋亡的影响。在合适药物浓度干预下,用蛋白免疫印迹法(Western blot)检测凋亡相关蛋白B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、剪切的胱天蛋白酶-3(cleaved Caspase-3)、胱天蛋白酶-3(Caspase-3)水平变化,同时测定缺氧诱导因子-1α(HIF-1α)及Hippo信号通路分子表达情况。结果:与空白组比较,芍药苷对人肺癌H1299、H292和A549细胞的生长均具有显著抑制作用(P<0.01);芍药苷可显著诱导A549细胞出现凋亡现象(P<0.01),同时降低Bcl-2/Bax(P<0.01),并显著升高cleaved Caspase-3蛋白的表达(P<0.01)。与空白组比较,芍药苷组A549细胞中的HIF-1α、具有PDZ基序的辅转录激活子(TAZ)、大肿瘤抑制激酶1(LATS1)、单极纺锤体结合蛋白1(MOB1)、Yes激酶相关蛋白(YAP)的表达水平均显著降低(P<0.01),p-LATS1、p-MOB1、p-YAP表达显著升高(P<0.01)。同时对磷酸化哺乳动物不育系20样激酶1(pMST1)及MST1的表达水平没有显著影响,差异无统计学意义。结论:芍药苷可通过改善细胞缺氧状态、抑制异常激活的Hippo信号通路诱导促进非小细胞肺癌凋亡,从而达到抗肿瘤的作用。
Objective::To observe the apoptosis induced by paeoniflorin(PF)in non-small cell lung cancer(NSCLC)cells and explore its mechanism.M ethod::Cell counting kit-8(CCK-8)was used to detect the inhibition rates of H1299,H292 and A549 cells with different concentrations of PF(2.5,5,10,20,25μmol·L-1),and to screen suitable concentrations of PF and experimental cells.The inhibitory effect of PF on lung cancer cells was detected by clone formation assay.The effect of PF on cell apoptosis was detected by flow cytometry with annexin V-FITC/propidium iodide(PI)double staining.With the right concentration of drugs,levels of apoptosis-associated protein B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),cleaved Caspase-3 and Caspase-3 were detected by Western blot.At the same time,the molecular expressions of hypoxia inducible factor-1α(HIF-1α)and Hippo signaling pathway were determined.Result::Compared with the blank group,PF significantly inhibited the growth of H1299,H292 and A549 cells of human lung cancer(P<0.01).PF significantly induced apoptosis in A549 cells(P<0.01),decreased the Bcl-2/Bax ratio(P<0.01),and significantly increased the cleaved Caspase-3 expression(P<0.01).Compared with those in the blank group,the expression levels of HIF-1α,transcriptional coactivator with PDZ-binding motif(TAZ),large tumor suppressor 1(LATS1),Mps one binding 1(MOB1)and Yes-associated protein(YAP)in A549 cells of the PF treatment group were significantly decreased(P<0.01),while the expressions of p-LATS1,p-MOB1 and p-YAP were significantly increased(P<0.01).At the same time,there was no significant effect on the expression levels of phosphorylated mammalian Ste20-like kinase 1(p-MST1)and MST1,which did not reach a statistical difference.Conclusion::All data demonstrated that PF showed an anti-tumor effect by improving hypoxic conditions and inhibiting the abnormally activated Hippo signaling pathway,thereby inducing and promoting apoptosis in non-small cell lung cancer.
作者
李燕
彭亮
蒋立峰
王生
王歌
于小林
姚玉琳
LI Yan;PENG Liang;JIANG Lifeng;WANG Sheng;WANG Ge;YU Xiaolin;YAO Yulin(The Affiliated Cancer Hospital of Zhengzhou University/Henan Cancer Hospital,Zhengzhou 450008,China;Fuwai Central China Cardiovascular Hospital/Henan Provincial People's Hospital,Zhengzhou 451464,China;Henan University of Chinese Medicine,Zhengzhou 450046,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2024年第12期39-44,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(82204844)
河南省科技攻关项目(222102310468)。
