EXPRESSION AND REVERSION OF DRUG RESISTANCE-AND APOPTOSIS-RELATED GENES OF A DDP-RESISTANT LUNG ADENOCARCINOMA CELL LINE

Objective: To investigate the co-expression of drug resistance- and apoptosis-related genes of cisplatin (CDDP)-selected lung adenocarcinoma cell line A 549 DDP for compared to the parental cell line A549, and reverse of drug resistance by antisense s-oligodeoxynucleotides (S-ODNs) of differentially expressed genes. Methods: Sense and antisense S-ODN were transferred into A 549 DDP cells by lipofectin. The expression of drug resistance and apoptosis related genes was examined by RT-PCR, immunocytochemistry and flow cytometry, respectively. Apoptostic cells were identified by DNA electrophoresis and terminal deoxynucleotidyl transferase (TdT)-mediated biotin dUTP nick end-labeling(TUNEL). Drug resistance of tumor cells was detected by a cell viability (MTT) assay. Results: The expression of bcl-2 was positive and that of multidrug resistance-associated protein (MRP) at mRNA and protein level was increased in A 549 DDP compared to A549 cells. MDR1, c-myc and topoisomeras II (TOPO II) were similarly co-expressed in two cell lines. Both cell lines were negative for c-erbB-2 expression. In A 549 DDP cells, the expression of bcl-2 and MRP was significantly inhibited by their respective antisense S-ODNs. Antisense S-ODNs could also decrease significantly drug resistance of A 549 DDP cells to CDDP by promoting cell apoptosis. Conclusion: Both intrinsic and acquired drug resistance were involved in co-expression of multiple MDR-related genes in lung adenocarcinoma. Cooperation of bcl-2 and MRP genes appeared to play an important action to confer the resistance of A 549 DDP cells to CDDP. Their antisense S-ODNs are responsible for the decrease of drug resistance of this cell line by promoting apoptosis.

Empirical studies about quercetin increasing chemosensitivity on human lung adenocarcinoma cell line A549

Objective： The present study was designed to investigate whether quercetin exerts increasing chemosensitivity on human lung adenocarcinoma cells when quercetin combined with cisplatin （DDP） and vincristine （VCR） in vitro respectively and its possible antitumor mechanism. To provide experimental proof for clinical combination application. Methods： Using intermittent administration of high dose VCR, human lung adenocarcinoma sensitive cell line （A549/S） was induced to VCR- resistant human lung adenocarcinoma cell line （A549NCR）. MTT assay was adapted for examing the 50% inhibition （IC50） value of DDP and VCR on A549/S and A549/VCR when quercetin combined with DDP and VCR respectively. Results： IC50 of DDP on A549/S and A549/VCR was 10.18 and 12.35 mg/L, and the IC50 of VCR on the two cell lines was 1.21 and 12.77 rag/L, respectively. The resistance fold of A549/VCR on VCR and DDP was 10.55 and 121, respectively. When quercetin at concentration of 50, 100 and 200 pmol/L in combination with DDP and VCR respectively, the IC50 of DDP and VCR on A549/S and A549/VCR were obvious decreased （P 〈 0.05 - P 〈 0.01）. Conclusion： The experiment results suggested that quercetin could increase the chemosensitivity and partly revise the resistance of A549NCR.

Reversal effect of recombinant human Endostatin on cisplatin resistance in A549/DDP human lung adenocarcinoma cells in vitro

Objective： Recombinant human Endostatin （rh-Endostatin, YH-16） can reverse cisplatin resistance in A549/DDP cells. However, the possible effect of rh-Endostatin in reversing DDP-resistance in A549/DDP cells and the mechanism are needed to be investigated. Methods： Lung adenocarcinoma cell line A549 and its DDP-resistant cell line A549/DDP were treated with DDP and/or recombinant human Endostatin. Difference in drug resistance was analyzed between different regi- mens and between different cell lines after a 72 h-treatment in vitro. And below the non-cytotoxic concentration of rh-End- ostatin, the possibility of rh-Endostatin in reversing DDP-resistance in A549/DDP was evaluated. The resistance protein which was detected in the study included P glycoprotein （P-gp） and topoisomerase II （Topo-II）. Results： Rh-Endostatin below 400 IJg/mL showed no cytotoxicity in either A549 or A549/DDP after 72 h-treatment with it. The inhibited concentration of 50% （IC50） observed for DDP was （0.79 _＋ 0.05） IJg/mL in A549 and （13.2 ＋ 1.1） in A549/DDP respectively. IC50 was reduced to 2.57 ＋ 0.05 #g/mL in A549/DDP treated by rh-Endostatin below the non-cytotoxic concentrations in combination with DDP, with a reversal fold （RF） of 5.14 and a relative reversal rate of 85.6%. Apoptotic rates were 2.01%, 13.47% and 29.26% re- spectively for cells treated with rh-Endostain, DDP, and the combination. The rate of the A549/DDP control group was 0.99%. The expression level of P-gp or Topo-II was higher in A549/DDP cells than in A549 cells. Rh-Endostatin may partially reverse DDP-resistance in A549/DDP cells in vitro, with a probable mechanism related to lowering expression of P-gp and Topo-II. Conclusien： Rh-Endostatin of non-cytotoxic dose partially reversed cisptatin resistance in cisplatin-resistant human lung adenocarcinoma cell line A549/DDP. Rh-Endostatin reversed the resistance of A549/DDP cells to DDP, which may be related to decreased protein expression of P-gp and Topo-II in A549/DDP cells.

染料木素对非小细胞型肺癌A549/DDP细胞增殖和凋亡的影响

目的:观察染料木素(genistein)对人非小细胞型肺癌(non small cell lung cancer,NSCLC)A549/DDP细胞增殖和凋亡的影响。方法:培养A549及A549/DDP细胞株,以A549细胞为对照。①MTT法测定A549及A549/DDP细胞对顺铂的IC50值、耐药倍数及细胞增殖抑制率;②测定0、1.25、2.5、5.0、10、20、40、60、80μg/ml染料木素作用48 h对A549/DDP细胞增殖的抑制率及IC50值;③用6.25、12.5、25μg/ml染料木素处理A549/DDP细胞24 h后,经流式细胞计量仪检测细胞周期及细胞凋亡情况。结果:①A549及A549/DDP细胞对顺铂的IC50值分别为33.6μmol/L和76.9μmol/L,耐药倍数为2.3;细胞增殖抑制率随顺铂浓度增加而逐渐加大;②染料木素对A549/DDP细胞生长的影响,随染料木素浓度增加表现为先促增殖后抑制的作用,其对A549及A549/DDP细胞的IC50值分别为85.1μg/ml和80.2μg/ml;③6.25、12.5、25μg/ml染料木素作用于A549/DDP细胞24 h后,随染料木素浓度的增加,停留于G2/M期的细胞数逐渐增多(P<0.05),同时A549/DDP细胞出现凋亡。结论:染料木素可抑制A549/DDP细胞的生长,将细胞阻滞于G2/M期,并诱导细胞凋亡。

橙皮甙对人非小细胞肺癌A549/DDP细胞增殖的影响

目的观察橙皮甙对耐顺铂的人非小细胞肺癌A549/DDP细胞增殖的影响。方法培养A549及A549/DDP细胞,绘制细胞生长曲线;MTT法测定A549/DDP细胞对顺铂的耐药倍数;②测定6、12、24、48及96 mg/L橙皮甙在6、12、24及48 h对A549/DDP细胞增殖的抑制率。③用24 mg/L橙皮甙处理A549/DDP细胞24 h后,用流式细胞计量术分析细胞周期。结果①A549/DDP细胞和A549细胞的生长曲线相似,倍增时间分别为(33.68±0.11)h和(33.54±0.84)h;A549/DDP细胞对顺铂的耐药倍数为12.90。②橙皮甙对A549/DDP细胞的抑制率随橙皮甙浓度增加和(或)时间的延长而增强(Pearson相关分析P<0.05)。③24 mg/L橙皮甙作用A549/DDP细胞24 h后,实验组G2/M期和S期细胞数比对照组减少(P<0.05)。结论①A549/DDP细胞基本保留了A549细胞的生长特性,对顺铂有明显耐药性。②橙皮甙对A549/DDP生长有抑制作用。③橙皮甙可诱导A549/DDP细胞阻滞于G2/M期。

川芎嗪对肺癌顺铂耐药细胞A549/DDP凋亡的影响

目的观察不同浓度川芎嗪(TMP)对肺癌顺铂耐药细胞株A549/DDP细胞的凋亡作用及其分子机制。方法应用不同浓度TMP(0.10、0.20、0.40、0.80、1.00mg·mL-1)处理A549/DDP细胞;噻唑蓝(MTT)比色法检测TMP对肺癌耐药细胞A549/DDP增殖抑制的影响,绘制细胞增殖抑制率曲线图;应用细胞流式技术检测A549/DDP的凋亡率;电子显微镜下观察A549/DDP细胞形态学特征的改变;提取A549/DDP细胞蛋白,并通过免疫印迹法检测应激活化蛋白激酶(JNK)、p38蛋白激酶、细胞外信号调节激酶(ERK)的蛋白表达变化。结果川芎嗪对A549/DDP细胞的增殖抑制率随着川芎嗪浓度的增加和作用时间的延长而逐渐增加,有时间和剂量依赖性(P<0.05)。TMP(0.20、0.40、0.80mg·mL-1)处理耐药细胞A549/DDP 72h后,细胞早期凋亡(AV+/PI-)比例分别为13.02%、26.58%、29.84%;电子显微镜下观察到TMP应用后肺癌耐药细胞A549/DDP出现凋亡特征性的凋亡小体;细胞蛋白免疫印迹实验表明,与对照组比较,TMP(0.20、0.40mg·mL-1)处理后A549/DDP细胞磷酸化(p)-p38[(72.03±5.27)、(86.63±5.09)比(47.24±3.64),P<0.05]和p-JNK的表达量[(143.79±3.88)、(163.02±6.35)比(130.18±5.37),P<0.05]增加而p-ERK表现出减少趋势[(62.33±5.61)、(24.00±4.89)比(108.52±6.24),P<0.05]。结论 TMP可能通过丝裂原活化蛋白激酶(MAPK)信号通路诱导肺癌耐顺铂细胞株A549/DDP凋亡。

Co-transfection of MRP and bcl-2 antisense S-oligodeoxynucleotides reduces drug resistance in cisplatin-resistant lung cancer cells

Obejctive To detect the influence of antisense s oligodeoxynucleotides (S ODNs) of bd 2 and multidrug resistamce associated protein (MRP) genes multidrug resistance associated protein gene and bcl 2 antisense S oligodeoxynucleotides on cisplatin resistant lung adenocarcinoma cell line A 549 DDP which overexpresses both bcl 2 and MRP Methods A 549 DDP cells were treated with sense and antisense S ODN mediated by lipofection Expression of MRP and bcl 2 mRNA and protein in the treated cells was measured by RT PCR and flow cytometry (FCM), respectively Apoptosis was identified by DNA electrophoresis and terminal deoxynucleotidyl transferase (TdT) mediated biotin dUTP nick end labeling(TUNEL) The degree of drug resistance of the treated cells was detected by a cell viability 3' [4,5 dimethylthiazol 2 yl] 2,5 diphenyl tefrazolium bromide thiazolylblue (MTT) assay Results Expression of bcl 2 and MRP significantly decreased in the cells treated with bcl 2 or/and MRP antisense S ODN for 48h as compared to the cells untreated and sense treated ( P <0 05) Resistance to cisplatin in the cells treated with bcl 2 or/and MRP antisense S ODN decreased by 60 6% (6 5 times), 56 4% (7 2 times) and 71 0% (4 8 times), respectively, which paralleled the decrease of bcl 2 and MRP expression Similarly, the resistance to etoposide and epirubicin in antisense treated cells also reduced in parallel to decreases of the two gene expressions The drug resistance in sense treated cells was similar to that in untreated cells Statistically significant dose and concentration dependent increases of apoptotic cells were observed in the groups exposed to 100?μmol/L cisplatin for 48?h after treatment by bcl 2 or/and MRP antisense Conclusion Bcl 2 and MRP were at least additive and possibly synergistic in conferring drug resistance in a cisplatin resistant lung adenocarcinoma cell line Antisense S ODN could attenuate drug resistance by promoting cells apoptosis, which might lead to a new treatment for patients with non small cell lung cancers (NSCLCs) who are refractory to conventional chemotherapy

THE EFFECTS OF THE ETHANOLIC FRACTION OF CORIOLUS VERSICOLOR ON THE A549 NON-SMALL-CELL LUNG CANCER CELL LINE

Coriolus versicolor has demonstrated anti-cancer effects via polysaccharide-peptides(PSP)and polysaccharide Krestin(PSK).However,many other bioactive compounds within Coriolus versicolor(CV)may not have been identified.Our primary focus was to determine whether the ethanolic extract of Coriolus versicolor demonstrated any anti-cancer effects.The crude ethanolic extract was utilized,as was

TGF-β/Akt/Smad signaling regulates ionizing radiation-induced epithelial-mesenchymal transition in acquired radioresistant lung cancer cells

Objective:To define the properties of lung cancer cells that resisted conventionally fractionated radiation exposure.Methods:Acquired radioresistant lung cancer cell line A549 was constructed by X-ray irradiation with a clinical conventional fraction dose of 2 Gy daily during 30 fractions.Cell morphology,molecular markers,migration capacity and invasion potential were evaluated by the microscope,Western blot,immunofluorescence,wound healing test and transwell chamber assay,respectively.Results:Radioresistant A549 cells shifted from an epithelial to a mesenchymal morphology,termed as epithelial-mesenchymal transition(EMT),and was accompanied by decreased expressions of epithelial markers(F=4.568,P<0.05)and increased expression of mesenchymal markers(F=4.270,P<0.05),greater migratory and invasive capabilities(t=6.386,5.644,P<0.05).The expression of TGF-β,and phosphorylated levels of Akt and Smad3 were also enhanced(F=6.496,4.685,3.370,P<0.05).Furthermore,the EMT phenotype induced by radiation could be reversed through inhibition of TGF-β,Akt or Smad3,indicating a functional relationship be-tween them.Conclusions:EMT mediates acquired radioresistance of lung cancer cells induced by IR with clinical parameters,and the crosstalk mode of TGF-β/Akt/Smad signaling plays a critical regulatory role in this process.

松节藻抗肿瘤活性

目的对松节藻中得到的10种化合物进行体外细胞毒活性筛选,并对其醇提取物进行体内抑瘤活性研究。方法采用碘酰罗丹明B(SRB)蛋白染色法和MTT法对10种化合物进行细胞毒活性筛选;用S180荷瘤小鼠模型进行松节藻醇提取物的体内抗肿瘤活性评价,分别测定半数致死量(LD50)、抑瘤率、胸腺指数、脾指数、脾淋巴细胞增殖活性(MTT法)、免疫球蛋白IgA、IgG、IgM和脾淋巴细胞凋亡率等指标,应用软件SPSS进行数据处理。结果体外活性筛选实验表明4,4′-亚甲基-二(5,6-二溴-1,2-二苯酚)()、3-溴-4-[2,3-二溴-4,5-二羟基苯基]甲基-5-(甲氧基甲基)-1,2-二苯酚()、3-溴-4,5-二(2,3-二溴-4,5-二羟基苯甲基)-1,2-二苯酚()、二(2,3-二溴-4,5-二羟基苯甲基)醚()和3-溴-4-[2,3-二溴-4,5-二羟基苯基]甲基-5-(乙氧基)-1,2-二苯酚()5种单体化合物对人肺癌细胞株A-549具有一定的细胞毒活性。松节藻醇提取物中剂量(1g/kg)组表现出较高的抑瘤率,但中剂量组胸腺指数及脾指数均有所升高;松节藻醇提取物各组均可较好抑制脾淋巴细胞增殖;各组免疫球蛋白IgA和IgG无明显变化,而松节藻醇提取物中剂量组IgM含量有明显升高;中、高剂量组脾淋巴细胞凋亡率显著升高。结论从松节藻中提取的部分化合物表现出一定的细胞毒活性;其醇提取物对肿瘤细胞有一定的抑制作用,并可增强机体免疫功能。

γ-干扰素与维拉帕米联合逆转人肺癌细胞系的多药耐药

目的 探讨逆转人肺癌耐药系HTB - 5 6R耐药性的可行性。方法 采用MTT比色法研究钙通道阻滞剂维拉帕米和人类重组γ -干扰素对DDP细胞毒性在人类肺癌细胞系HTB - 5 6R上的的影响。结果 γ -干扰素与维拉帕米能部分恢复HTB - 5 6R对顺铂 (DDP)的敏感性。当维拉帕米浓度 >4mg/L时 ,能明显增加DDP的细胞毒性 ,耐药株的抑制率 >19.0 % (P <0 .0 5 )。γ -干扰素浓度 >4.0× 10 6 U/L时 ,也能达到此效果 ,细胞抑制率 >3 6.9% (P <0 .0 5 )。联合两药比单独使用有更好的效果 ,维拉帕米为 2mg/L、γ -干扰素为 2 .0× 10 6 U/L时 ,细胞抑制率就达 3 6.9% (P <0 .0 1)。不论单独使用还是联合使用逆转效果均呈剂量效应。结论 γ -干扰素与维拉帕米能逆转HTB - 5 6R的耐药性 ,而且联合使用有更好的效果。

RASSF1A逆转A549-DDP细胞对顺铂耐受的实验研究

RASSF1A表达下降可能是肺癌对顺铂获得性耐药的主要因素之一。为了寻找预测顺铂对肺腺癌A549细胞化疗敏感性的生物标志,该研究以对顺铂耐药的同源A549细胞(A549-DDP)为研究对象,利用转染技术上调RASSF1A表达。RT-PCR和Western blot检测转染前后RASSF1A在mRNA和蛋白水平的表达情况。在不同浓度顺铂作用下,通过MTT法、克隆形成实验检测转染前后细胞活力并计算IC50和计数克隆形成数。在同一浓度顺铂作用下,流式细胞仪检测转染前后细胞凋亡分数的改变。结果显示:顺铂作用24 h后,转染RASSF1A表达质粒的A549-DDP细胞对顺铂IC50明显低于A549-DDP细胞[(39.9±6.3)μmol/L vs(53.0±5.8)μmol/L,P=0.036];不同浓度顺铂连续作用5天后,转染RASS-F1A表达质粒的A549-DDP细胞形成的克隆数明显少于A549-DDP细胞组;在同一浓度顺铂作用下,转染RASSF1A表达质粒的A549-DDP凋亡分数明显大于A549-DDP细胞[(7.10±0.01)%vs(3.80±0.18)%,P=0.002]。结果提示,RASSF1A表达下降可能是肺腺癌A549细胞对顺铂获得性耐药的重要因素之一,RASSF1A表达在临床上作为肺癌患者顺铂化疗耐受的生物标志值得进一步研究。

奥美拉唑逆转人肺腺癌耐顺铂细胞株A549/DDP耐药性的研究

目的:探讨液泡ATPase[vacuolar(H+)-ATPase,V-ATPase]非特异性抑制剂奥美拉唑(omeprazole,OME)逆转人肺腺癌耐药细胞株A549/DDP的耐药性及可能的作用机制。方法:以非细胞毒性浓度4μg/mL的OME预处理A549/DDP细胞24 h,再用2μg/mL顺铂(cisplatin,DDP)处理A549/DDP细胞。MTT法检测细胞的增殖抑制率,激光扫描共聚焦显微镜(laser scanning confocal microscope,LSCM)法间接检测细胞内pH值的变化,FCM法检测凋亡相关蛋白Bcl-2和PTEN的表达,RT-PCR法检测VATPase、Bcl-2和PTEN mRNA的表达。结果:OME具有逆转A549/DDP细胞耐药性的作用,逆转倍数为1.45(P<0.01);OME预处理后A549/DDP细胞内pH值明显降低,Bcl-2蛋白表达下降,PTEN蛋白表达增加(P<0.01)。V-ATPase在亲本A549及A549/DDP细胞中相对表达量分别为0.88±0.00和0.99±0.00(P<0.01);A549/DDP细胞中V-ATPase在有无OME预处理的情况下的相对表达量分别为1.09±0.00和1.05±0.02,差异无统计学意义。Bcl-2 mRNA相对表达量下降(P<0.01),PTEN mRNA相对表达量增加(P<0.01)。结论:V-ATPase在A549/DDP细胞中高表达,OME能部分逆转A549/DDP耐药性,其作用机制可能与上调PTEN和下调Bcl-2表达有关。

3-BrPA对肺癌A549/DDP细胞耐药的逆转作用研究

目的探究3-Br PA对肺癌A549/DDP细胞耐药的逆转作用及其机制。方法 1细胞培养:RPMI1640培养基培养肺癌A549细胞及人肺腺癌耐顺铂细胞株A549/DDP细胞;2采用CCK8及Western blot法观察3-Br PA对A549/DDP细胞的耐药逆转作用和P-糖蛋白(P-glycoprote,P-gp)的表达水平;采用流式细胞术检测不同浓度3-Br PA作用A549/DDP细胞后细胞内罗丹明-123(Rhodamine-123,Rh123)的蓄积情况。结果顺铂对A549细胞、A549/DDP细胞及3-Br PA作用后的A549/DDP细胞的IC50分别为6.714μg/ml、38.99μg/ml及12.86μg/ml,逆转倍数为3.03,相对逆转效率为81%。3-Br PA可使A549/DDP细胞的P-gp表达下调。3-Br PA可使A549/DDP细胞内的Rh123蓄积增多并呈剂量依赖性。结论 3-Br PA可逆转A549/DDP细胞对顺铂耐药,作用机制可能与其抑制P-gp的功能和表达有关。

Agglutinin isolated from Arisema heterophyllum Blume induces apoptosis and autophagy in A549 cells through inhibiting PI3K/Akt pathway and inducing ER stress

Arisaema heterophyllum Blume is one of the three medicinal plants known as traditional Chinese medicine Rhizoma Arisaematis(RA). RA has been popularly used to treat patients with convulsions, inflammation, and cancer for a long time. However, the underlying mechanisms for RA effects are still unclear. The present study was designed to determine the cytotoxicity of agglutinin isolated from Arisema heterophyllum Blume(AHA) and explore the possible mechanisms in human non-small-cell lung cancer A549 cells. AHA with purity up to 95% was isolated and purified from Arisaema heterophyllum Blume using hydrophobic interaction chromatography. AHA dose-dependently inhibited the proliferation of A549 cells and induced G_1 phase cell cycle arrest. AHA induced apoptosis by up-regulating pro-apoptotic Bax, decreasing anti-apoptotic Bcl-2, and activating caspase-9 and caspase-3. In A549 cells treated with AHA, the PI3K/Akt pathway was inhibited. Furthermore, AHA induced increase in the levels of ER stress markers such as phosphorylated eukaryotic initiation factor 2α(p-eIF2α), C/EBP-homologous protein(CHOP), inositol-requiring enzyme 1α(IRE1α), and phosphorylated c-Jun NH_2-terminal kinase(p-JNK). AHA also induced autophagy in A549 cells. Staining of acidic vesicular organelles(AVOs) and increase in the levels of LC3II and ATG7 were observed in AHA-treated cells. These findings suggested that AHA might be one of the active components with anti-cancer effects in Arisaema heterophyllum Blume. In conclusion, cytotoxicity of AHA on cancer cells might be related to its effects on apoptosis and autophagy through inhibition of PI3K/Akt pathway and induction of ER stress.

Synthesis, Characterization and Biological Screening of Ferulic Acid Derivatives

According to WHO, cancer is a leading cause of death worldwide, accounting for 8.2 million deaths in 2012. Among several factors involved in the pathogenesis of cancer, free radical formation followed by damage to DNA and cell protein is one of the causes. Natural plant products have gained enormous interest in the prevention or treatment of chronic diseases. Ferulic acid, like other phenolic acids (caffeic acid, sinapic acid) possess anti cancer activity. A series of ferulic esters (FE1 - FE11) and ferulic amides (FA1 - FA10) were synthesized and evaluated for their cytotoxic activity.

小白菊内酯对人肺癌A-549细胞周期变化和凋亡通路的影响

目的:研究小白菊内酯对人肺癌A-549细胞株的抗增殖和诱导凋亡作用及其机制,为研发新型抗肿瘤中药奠定坚实的基础。方法:应用MTT法测定小白菊内酯对肺癌细胞的毒性作用和抑制增殖作用;应用倒置显微镜、Annexin V-FITC/碘化丙锭(PI)染色荧光显微镜观察药物作用前后人肺癌A-549细胞的形态学变化;应用活性氧(ROS)检测药物作用后细胞内活性氧的水平,以及活性氧含量与细胞凋亡之间的关系;应用免疫细胞化学技术检测药物作用前后蛋白阳性表达变化的情况并进一步探讨小白菊内酯对人肺癌A-549细胞株抑制增殖和诱导凋亡的作用机制。结果:(1)MTT法检测结果显示:小白菊内酯在不同浓度(5mg/L、10 mg/L、20 mg/L、40 mg/L、50 mg/L、55 mg/L、60 mg/L、65 mg/L)对人肺癌A-549细胞有不同程度的生长抑制作用,并呈现出浓度和时间梯度依赖性,当药物终浓度为55 mg/L作用时间为48h时,对A-549细胞的抑制率为51.02%。(2)Annexin V-FITC/PI染色荧光显微镜观察发现实验组细胞可见大量绿色荧光细胞和红色凋亡细胞。(3)活性氧检测发现:实验组细胞内活性氧的水平出现明显升高。(4)免疫细胞化学检测法结果显示对照组Caspase-3、Caspase-8、Caspase-9、Fas和Bax分别为10.07%、11.42%、14.09%、9.75%和11.87%,加入小白菊内酯后(实验组)细胞中Caspase-3、Caspase-8、Caspase-9、Fas和Bax蛋白的表达为89.24%、85.75%、84.46%、85.14%和84.09%,与对照组相比明显增加,有显著差异。结论:小白菊内酯作用于人肺癌A-549细胞株后,诱导其出现细胞凋亡的改变,它通过死亡受体通路和线粒体通路来完成诱导凋亡的机制,小白菊内酯作用后活性氧水平的增加可能参与诱导细胞凋亡的机制。