厄罗替尼联合塞来昔布阻断EGFR和COX-2抑制肺癌A549细胞增殖

目的探讨厄罗替尼联合塞来昔布对肺腺癌A549细胞系凋亡、表皮生长因子受体(EGFR)和环氧合酶-2(COX-2)表达的影响。方法厄罗替尼、塞来昔布单独或联合干预细胞48 h后,倒置相差显微镜观察细胞形态;四甲基偶氮唑盐(MTT)法测定细胞抑制率;Hoechst33258法和TUNEL法检测细胞凋亡和流式细胞术检测细胞周期;免疫荧光检测EGFR和COX-2蛋白表达。结果厄罗替尼联合塞来昔布组相比单药组A549细胞明显出现大量颗粒和空泡,细胞变圆开始脱落;二者联合作用时抑制作用更强(P<0.05),厄罗替尼与塞来昔布均能诱导细胞凋亡,联合作用后细胞凋亡率更高(P<0.05),并使细胞发生明显的G1期阻滞(P<0.05),进一步下调了EGFR和COX-2蛋白的表达(P<0.05)。结论厄罗替尼与塞来昔布联合应用可协同介导细胞凋亡,阻滞细胞周期于G0/G1期及阻滞EGFR和COX-2信号途径。     

NF-κB圈套寡核苷酸促进肺癌细胞A549凋亡的作用

目的:观察NF-κB圈套寡核苷酸对NF-κB活性的影响,研究其在肺癌细胞A549凋亡中的作用机制。方法: 用转染技术将FITC标记的NF-κB圈套寡核苷酸转入人肺癌细胞A549中,通过荧光倒置显微镜和共聚焦显微镜进行核内定位的观察,凝胶迁移率实验(EMSA)检测转染前后NF-κB活性的变化。生长曲线观察低活性NF-κB对细胞增殖的影响。流式细胞仪、TUNEL检测细胞凋亡率。Western blot检测NF-κB低活性引起的凋亡相关蛋白Bcl-2和Fas的改变。结果: NF-κB圈套寡核苷酸转染后定位于细胞核内,EMSA显示转染后NF-κB活性明显下降。A549细胞对NF-κB圈套寡核苷酸敏感,细胞生长速度减缓。转染NF-κB圈套寡核苷酸的A549细胞凋亡率增加。NF-κB 的低活性可以引起凋亡抑制蛋白Bcl-2表达水平下调和凋亡蛋白Fas表达增加。结论: NF-κB圈套寡核苷酸具有促进肺癌细胞A549凋亡的作用,其机制可能与NF-κB圈套寡核苷酸通过下调NF-κB的活性使凋亡蛋白Fas表达增加和凋亡抑制蛋白Bcl-2表达降低有关。     

盐霉素增强吉非替尼诱导人肺腺癌细胞株A549凋亡的作用

 目的：探讨盐霉素联合吉非替尼诱导人肺腺癌细胞株A549凋亡的协同作用。方法：采用MTT的方法检测盐霉素对A549细胞生长的抑制作用;流式细胞术检测盐霉素对A549细胞凋亡和线粒体膜电位的影响;比色法检测caspase-3、-8和-9活性;Western bloting 分析细胞色素C、Bcl-2、p-EGFR、p-Akt和p-ERK蛋白水平。结果：盐霉素与吉非替尼单用均出现不同程度的细胞增殖抑制作用和诱导细胞凋亡作用;而盐霉素与吉非替尼联合作用,能更显著地抑制细胞增殖,且凋亡细胞显著增加（P<0.05）。盐霉素单独作用A549细胞,线粒体膜电位显著下降,细胞内活性氧和Ca2+在短期内显著升高,胞浆细胞色素C含量以及caspase-3、-8和-9活性均显著增加,与对照组比较差异均有统计学意义;吉非替尼单用则主要表现为对p-EGFR、p-Akt和p-ERK蛋白表达的抑制作用,而对胞浆细胞色素C含量以及caspase-3、-8和-9活性影响较少。Western blotting检测发现,联合用药组的Bcl- 2、p-EGFR、p-Akt和p-ERK蛋白表达量明显减少,但是对EGFR、Akt和ERK总蛋白水平无显著影响。结论：盐霉素与吉非替尼联用具有较好的协同作用,可能通过Bcl-2途径及线粒体凋亡途径诱导人肺腺癌A549细胞凋亡,提高A549细胞对吉非替尼的敏感性。     

伊马替尼诱导原代T细胞凋亡作用机制研究

目的：研究伊马替尼（IM）诱导正常人原代CD3⁺ T细胞凋亡的分子机制。 方法： 0～100 nmol/L的IM作用原代T细胞24 h后,应用流式细胞术检测细胞凋亡情况;应用实时荧光定量PCR技术检测Caspase-3、Caspase-8、A20和NF-κB基因表达变化;应用Western blot技术检测A20和NF-κB蛋白表达变化。结果：IM具有明显促T细胞凋亡能力,其凋亡分子Caspase-3 和A20基因表达水平均上调,而NF-κB的基因和蛋白水平均下调。结论：IM通过上调A20的表达诱导了T细胞的凋亡。     

泰素帝对肺腺癌细胞A549凋亡及Survivin表达和Caspase-3活性的影响

目的探讨泰素帝对肺腺癌细胞A549凋亡,生存素(Survivin)表达和Caspase-3活性的影响。方法体外细胞培养,待细胞生长处于对数期时,加入不同浓度泰素帝,MTY比色法检测细胞活性。参考MTT结果,选取100ng/ml泰素帝处理A549细胞。透射电镜,流式细胞术检测细胞凋亡的发生;RT-PCR检测Survivin mRNA的表达;Western印迹法检测Survivin蛋白质的表达;Caspase-3活性检测试剂盒检测Caspase-3活性。结果泰素帝可抑制A549细胞生长,诱导细胞凋亡,呈时间依赖性降低Survivin的表达,增强Caspase-3酶活性。结论 100ng／ml泰素帝可诱导A549细胞凋亡,Survivin表达降低和Caspase-3活性增强可能是其诱导肺腺癌细胞凋亡的分子途径之一。     

NF-κB介导MRP8/14诱导的人肺泡上皮A549细胞凋亡

 目的: 探讨髓样相关蛋白8/14(MRP8/14)对人肺泡上皮A549细胞存活及凋亡的影响,并研究核因子κB(NF-κB)在其中的作用。方法: MRP8/14以不同剂量或刺激时间处理A549细胞,CCK-8法检测其对细胞活力的影响;流式细胞术检测细胞凋亡率;Western blot法及间接免疫荧光法检测A549细胞内NF-κB p65蛋白核移位情况;Western blot法检测NF-κB p65蛋白磷酸化水平;使用NF-κB特异性抑制剂Bay 11-7082探讨NF-κB在凋亡过程中的作用。结果: MRP8/14能以剂量和时间依赖的方式影响A549细胞的存活(P<0.05),流式细胞技术检测结果表明MRP8/14处理后A549细胞凋亡率明显增加(P<0.01)。此外,MRP8/14能够诱导NF-κB p65蛋白发生显著磷酸化,并移位至细胞核中,NF-κB信号通路明显激活,而NF-κB特异性抑制剂Bay 11-7082能够明显抑制MRP8/14诱导的细胞凋亡(P<0.01)。结论: NF-κB在MRP8/14所导致的肺泡上皮细胞凋亡过程发挥了重要作用。     

安罗替尼通过调控miR-16-5p/PD-1轴抑制人非小细胞肺癌细胞系增殖并促进凋亡

目的 探讨安罗替尼对非小细胞肺癌细胞增殖和凋亡的影响及其分子机制。方法 将非小细胞肺癌细胞系A549和H1299分别采用安罗替尼、miR-16-5p激动剂和/或PD-1过表达载体进行处理。CCK-8实验和EDU实验检测细胞增殖;流式细胞术检测细胞凋亡;RT-qPCR检测miR-16-5p相对表达量;Western blot检测程序性细胞死亡-1蛋白(PD-1)的表达。双荧光素酶报告实验确定miR-16-5p和PD-1的靶向关系。用A549细胞构建裸鼠成瘤模型,检测安罗替尼对体内肿瘤生长的影响。结果 安罗替尼在A549和H1299细胞中以剂量依赖的方式显著增加miR-16-5p表达,同时降低PD-1表达,并且抑制细胞增殖,促进细胞凋亡(P<0.05)。miR-16-5p过表达可抑制细胞增殖,促进细胞凋亡(P<0.05)。miR-16-5p能靶向PD-1,且负调控PD-1表达。siRNA下调PD-1表达后明显抑制细胞增殖,并促进细胞凋亡(P<0.05)。过表达PD-1则可逆转安罗替尼介导的miR-16-5p对A549和H1299细胞的促增殖和抗凋亡作用(P<0.05...     

抑制抗增殖蛋白2增强非小细胞肺癌细胞系A549对厄洛替尼敏感性

目的 探讨抗增殖蛋白2(PHB2)在非小细胞肺癌细胞系A549对厄洛替尼(Erl)敏感性中的作用及其机制。方法 在A549细胞中转染PHB2小干扰RNA(siPHB2),观察抑制PHB2表达对Erl诱导的细胞增殖和凋亡的影响。利用MitoTracker染色和感染绿色荧光蛋白-微管相关蛋白(GFP-LC3)观察微管相关蛋白(LC3)和线粒体共定位,EdU实验检测细胞增殖,平板集落检测细胞集落形成能力,TUNEL实验检测细胞凋亡,Western blot检测PHB2和LC3Ⅱ蛋白表达水平,用相应试剂盒检测线粒体膜电位、细胞色素c含量和呼吸链复合物Ⅰ/Ⅱ/Ⅴ活性。结果 与siPHB2组和siCtrl+Erl组相比,siPHB2+Erl组EdU阳性的细胞数显著减少(P<0.05),集落形成数显著减少(P<0.05),TUNEL阳性的细胞数显著增加(P<0.05),线粒体膜电位显著降低(P<0.05),线粒体呼吸链复合物Ⅰ/Ⅱ/Ⅴ活性均显著降低(P<0.05),线粒体内细胞色素c减少(P<0.05),而细胞质内细胞色素c的增加(P<0.05)。结论 抑制...     

赫赛汀对吉非替尼诱导肺癌A549细胞凋亡的影响

目的： 研究吉非替尼与赫赛汀联合应用对人肺腺癌A549 细胞凋亡的影响。方法: 应用MTT法,流式细胞仪Annexin V－PI 双标法、DAPI荧光染色等多项方法,体外研究吉非替尼与赫赛汀联合对A549 细胞的促凋亡作用。结果: 吉非替尼与赫赛汀单独应用及联合应用均可以明显抑制人肺腺癌A549细胞的生长，促进细胞凋亡,并呈浓度及时间依赖性,2者联合作用人肺腺癌A549 细胞24 h、48 h、72 h 的凋亡率显著高于单用吉非替尼或赫赛汀组(P<0.05),2者呈现出相加的抗瘤效果。结论: 吉非替尼与赫赛汀联合应用在体外对人肺腺癌A549 细胞有明显的促凋亡作用。     

肝再生磷酸酶-3抗体联合培美曲塞促进人肺腺癌细胞系A549的凋亡

目的探讨肝再生磷酸酶-3(PRL-3)抗体和培美曲塞(Pem)单用及联用对人肺腺癌细胞系A549增殖、迁移和凋亡的影响。方法不同浓度PRL-3抗体处理A549细胞;CCK-8法测细胞增殖抑制率;Transwell小室法检测细胞迁移能力;流式细胞仪检测细胞凋亡率;Western bolt检测PRL-3、磷酸化细胞外信号调节激酶(P-ERK)及血管内皮生长因子(VEGF)蛋白表达。结果1)与对照组相比,PRL-3抗体随浓度增加,对A549细胞增殖的抑制作用增加(P<0.05);2)联合用药组对A549细胞增殖和迁移能力的抑制作用优于单药组(P<0.05);3)联合用药组A549细胞凋亡率34.63%±1.65%高于PRL-3抗体组17.24%±1.15%和培美曲塞组15.32%±1.82%(P<0.05);4)与单药组相比,联合用药组PRL-3、P-ERK及VEGF蛋白表达下降更显著(P<0.05)。结论PRL-3抗体联合培美曲塞可抑制A549细胞增殖、迁移并诱导凋亡,效果优于单药。其作用机制可能与抑制PRL-3、P-ERK和VEGF蛋白表达相关。     

HIF-1α介导了间歇低氧对人肺腺癌A549细胞体外生物学行为的影响

目的:探讨缺氧诱导因子1α(HIF-1α)是否介导间歇低氧对人肺腺癌A549细胞体外活力、凋亡以及侵袭的影响。方法:采用体外转染方法将特异性针对HIF-1α的siRNA导入A549细胞,在间歇低氧条件下培养后通过real-time PCR和Western blot法检测HIF-1α及其下游Bcl-2、Bax、P53、P21、VEGF的mRNA和蛋白表达;通过MTT法和流式细胞术分别检测A549细胞的活力、凋亡及细胞周期;通过Transwell小室检测A549细胞的侵袭能力。结果:未转染HIF-1α-siRNA的A549细胞[间歇低氧空白对照组(IHC组)、间歇低氧空载体对照组(IHE组)及间歇低氧阴性对照组(IHN组)]经间歇低氧干预后的HIF-1α、Bcl-2及VEGF表达均明显高于常氧对照组(RA组),Bax及P21表达均明显低于RA组(P0.05),而转染HIF-1α-siRNA的A549细胞[间歇低氧siRNA组(IHS组)]较IHC组、IHE组及IHN组经间歇低氧干预后的HIF-1α、BCL-2及VEGF表达均明显下调,Bax及P21表达较均明显上调(P0.05);所有间歇低氧组A549细胞的P53表达均较RA组升高(P0.05),但各间歇低氧组之间无显著性差异。IHC组、IHE组及IHN组A549细胞经间歇低氧干预后较RA组的细胞活力增强,凋亡率下降,侵袭力增强(P0.05),而IHS组A549细胞经间歇低氧干预后细胞周期被阻滞于G1期,较未转染组的细胞活力下降,凋亡增加,侵袭能力下降(P0.05)。结论:间歇低氧可通过HIF-1α途径调控其下游基因表达进而促进A549细胞的活力和转移;通过RNA干扰技术造成HIF-1α基因沉默可以抑制间歇低氧引起的A549细胞生长和转移。     

低氧诱导的小窝蛋白-1上调参与人肺腺癌细胞A549迁移和侵袭

 目的:探讨低氧诱导剂二氯化钴（CoCl2）对小窝蛋白-1（Cav-1）生成的调节作用及后者对人肺腺癌A549细胞迁移、侵袭的影响。方法:检测肺癌患者伴恶性胸水（MPE）和结核性胸膜炎患者胸水（TBPE）中Cav-1和缺氧诱导因子(HIF)-1α浓度,比较两者相关性;以CoCl2和（或）HIF-1α抑制剂YC-1作用于A549细胞ELISA法检测细胞上清Cav-1和HIF-1α浓度;分别采用细胞划痕实验及Transwell小室侵袭实验研究CoCl2刺激表达的Cav-1对A549细胞迁移和侵袭的影响。结果:MPE中Cav-1和HIF-1α浓度明显高于TBPE,两组患者胸水中Cav-1与HIF-1α均呈正相关。CoCl2浓度和时间依赖性诱导A549细胞Cav-1和HIF-1α生成,200 μmol/L或24 h达到峰值;浓度>200 μmol/L或作用时间超过24 h则呈现浓度或时间依赖性抑制。HIF-1α抑制剂YC-1浓度依赖性抑制HIF-1α和Cav-1生成。CoCl2浓度依赖性增强A549细胞迁移和侵袭,200 μmol/L作用最强;YC-1对上述过程产生抑制效应。结论: 肺癌患者胸腔积液中Cav-1浓度升高,低氧诱导Cav-1生成的变化可能参与了A549细胞迁移和侵袭,HIF-1α可能对Cav-1生成发挥影响。     

Human Apurinic/Apyrimidinic Endonuclease siRNA Inhibits the Angiogenesis Induced by X-Ray Irradiation in Lung Cancer Cells

Objective: Radiotherapy is an important and effective treatment method for non-small cell lung cancer (NSCLC). Nonetheless, radiotherapy can alter the expression of proangiogenic molecules and induce angiogenesis. Human apurinic/apyrimidinic endonuclease (APE1) is a multifunctional protein, which has DNA repair and redox function. Our previous studies indicated APE1 is also a crucial angiogenic regulator. Thus, we investigated the effect of APE1 on radiation-induced angiogenesis in lung cancer and its underlying mechanism.Methods: Tumor specimens of 136 patients with NSCLC were obtained from 2003 to 2008. The APE1 and vascular endothelial growth factor (VEGF) expression, as well as microvessel density (MVD) were observed with immunohistochemistry in tumor samples. Human lung adenocarcinoma A549 cells were treated with Ad5/F35-APE1 siRNA and/or irradiation, and then the cells were used for APE1 analysis by Western blot and VEGF analysis by RT-PCR and ELISA. To elucidate the underline mechanism of APE1 on VEGF expression, HIF-1α protein level was determined by Western blot, and the DNA binding activity of HIF-1α was detected by EMSA. Transwell migration assay and capillary-like structure assay were used to observe the migration and capillary-like structure formation ability of human umbilical veins endothelial cells (HUVECs) that were co-cultured with Ad5/F35-APE1 siRNA and (or) irradiation treated A549 cells culture medium.Results: The high expression rates of APE1 and VEGF in NSCLC were 77.94% and 66.18%, respectively. The expressions of APE1 was significantly correlated with VEGF and MVD (r=0.369, r=0.387). APE1 and VEGF high expression were significantly associated with reduced disease free survival (DFS) time. The high expressions of APE1 and VEGF on A549 cells were concurrently induced by X-ray irradiation in a dose-dependent manner. Silencing of APE1 by Ad5/F35-APE1 siRNA significantly decreased DNA binding activity of HIF-1α and suppressed the expression of VEGF in A549 cells, moreover, significantly inhibited the endothelial cells immigration and capillary-like structure formation induced by irradiated A549 cells.Conclusion: Our results indicate that APE1 may play a crucial role in angiogenesis induced by irradiation. Administration of Ad5/F35-APE1 siRNA during radiotherapy could be a potent adjuvant therapeutic approach to enhance the radiotherapy response, effectively eliminate metastasis and improve the efficacy of radiotherapy for NSCLC.     

Inhibition of ovarian cancer proliferation and invasion by pachymic acid

To determine the effect of pachymic acid (PA) on proliferation, cell cycle, and invasion in human ovarian carcinoma cell lines HO-8910 and explore some possible mechanisms, HO-8910 cells was treated with different concentrations of PA (0.5, 1, 2 μM). CCK-8 assay, propidium iodide staining, was applied to measuring the growth inhibiting rates of HO-8910 cells. Cell cycle was measured by flow cytometry. In addition, the activity of PA against HO-8910 cells invasion was evaluated in transwell assay. Western blot detected the proteins expression of E-cadherin, β-catenin and COX-2 of different groups treated with PA in different concentrations (0.5, 1, 2 μM) for 48 h. Our results showed that PA could effectively inhibit the in vitro growth of HO-8910 cells in dose-dependent manners in 72 h, suppressed migration and invasion of HO-8910 cells in concentration-dependent manners at 24 h, caused the increased accumulation of G1 phase cells, and caused down-regulation of β-catenin and COX-2 and up-regulation of E-cadherin expression level. Taken together, it could conclude that PA might inhibit proliferation and invasion of ovarian carcinoma cell through decreasing β-catenin and COX-2 expression and increasing E-cadherin exprssion.     

BHD基因在人肺腺癌A549细胞的表达及其对A549细胞转移能力的影响

目的 了解BHD基因在肺癌细胞中的表达部位及其在影响肺癌细胞转移、运动方面的作用. 方法 制备BHD真核表达质粒,转染肺腺癌细胞,用免疫荧光法检测Folliculin蛋白在肺癌细胞中的表达位置;筛选sRNAi片段,选取有效抑制片段转入肺癌细胞,与转入BHD质粒的肺癌细胞对比,用Transwell实验了解BHD基因对肺癌细胞运动方面的影响. 结果 转入BHD质粒后,A549细胞质内出现明显的Folliculin蛋白表达,Transwell实验显示,转入BHD质粒能抑制A549细胞的侵袭能力,而转入BHD siRNA片段的A549细胞运动迁移能力升高.结论 BHD表达产物Folliculin蛋白在肺癌细胞的胞质内表达,BHD可能是肺癌转移抑制基因.     

HIF-2α、VEGF、COX-2、MMP-9表达及其与肾细胞癌血管生成的关系分析

目的 探讨HIF-2α(缺氧诱导因子-20α)、VEGF(血管内皮生长因子)、COX-2(环氧化酶-2)、MMP-9(金属基质酶-9)表达及其与肾细胞癌血管生成的关系.方法 应用免疫组化方法,检测79例肾细胞癌手术切除标本的HIF-2α、VEGF、COX-2、MMP-9表达情况,并与MVD(微血管密度)表达进行关联性分析.结果 肾癌组织MMP-9、VEGF的表达均显著高于正常肾组织(P＜0.05);不同临床分期肾癌组织中的MMP-9、VEGF表达有明显差异(P＜0.05).MVD与VEGF、MMP-9蛋白表达呈显著正相关(r=0.381、0.375,P＜0.05);MVD与COX-2表达有关,COX-Z表达0级和4级MVD值最低,与1、2、3级之间比较差异有统计学意义(P＜0.05),0级与4级之间比较差异无统计学意义(P＞0.05),1、2、3级之间差别无统计学意义(P＞0.05).肾细胞癌中HIF-2α阳性组的MVD值高于HIF-2α阴性组中MVD值,差异有统计学意义(t=4.374,P＜0.05);Spearman等级相关分析发现,HIF-2α的表达与MVD间存在正相关关系(r=0.545,P＜0.01).结论 HIF-2α、VEGF、COX-2、MMP-9表达及其与肾细胞癌血管生成之间有明显相关性,在临床上可以根据相关基因表达情况而采取相应的干预措施.     

Effects of Coptis extract combined with chemotherapeutic agents on ROS production, multidrug resistance, and cell growth in A549 human lung cancer cells

ABSTRACT: BACKGROUND: Non-small cell lung cancer is associated with high expression of multidrug resistance (MDR) proteins and low production of reactive oxygen species (ROS). Coptis extract (COP), a Chinese medicinal herb, and its major constituent, berberine (BER), have anticancer properties. This study aims to investigate the effects of COP and BER combined with chemotherapeutic agents, including fluorouracil (5-FU), camptothecin (CPT), and paclitaxel (TAX) on cell proliferation, ROS production, and MDR in A549 human non-small cell lung cancer cells. METHODS: A549 cells were treated with different doses of COP and BER, combined with 5-FU, CPT, and TAX. Cell viability was measured by an XTT (2,3-bis-(2-methoxy-4- nitro-5-sulfophenyl)-2 H-tetrazolium-5-carboxanilide) assay. Intracellular ROS levels were determined by measuring the oxidative conversion of cell permeable 2' ,7' -dichlorofluorescein diacetate to fluorescent dichlorofluorescein. MDR of A549 cells was assessed by rhodamine 123 retention assay. RESULTS: Both COP and BER significantly inhibited A549 cell growth in a dose-dependent manner. Combinations of COP or BER with chemotherapeutic agents (5-FU, CPT, and TAX) exhibited a stronger inhibitory effect on A549 cell growth. In addition, COP and BER increased ROS production and reduced MDR in A549 cells. CONCLUSION: As potential adjuvants to chemotherapy for non-small cell lung cancer, COP and BER increase ROS production, reduce MDR, and enhance the inhibitory effects of chemotherapeutic agents on A549 cell growth.     

RETRACTED: Resibufogenin suppresses growth and metastasis through inducing caspase-1-dependent pyroptosis via ROS-mediated NF-κB suppression in non-small cell lung cancer

The purpose of this study is to explore the antitumor properties of resibufogenin (RB) in non-small cell lung cancer (NSCLC) and elucidate its underlying mechanism. A549 and H520 cells were treated with various concentrations of RB with or without NLRP3 inhibitor (MCC950), caspase-1 inhibitor (VX765), or N-acetyl-l -cysteine (an ROS scavenger). Cell counting kit-8 and colony formation assays were conducted to determine cell viability. Cell invasion was detected by using the transwell assay. The release of lactate dehydrogenase (LDH) was determined by the LDH detection assay. The protein expression levels of related genes were measured by western blotting and immunohistochemistry. The reactive oxygen species (ROS) level was detected by using a 2,7-dichlorodihydrofluorescein diacetate ROS Assay Kit. The in vivo effects of RB were evaluated in a xenograft mouse model. RB treatment reduced cell viability and invasion in a dose-dependent manner. Furthermore, RB also enhanced pyroptosis levels in A549 and H520 cells, as indicated by the increased release of LDH and pyroptosis-related proteins. Interestingly, we also found that the antiproliferative and antimetastatic effects of RB were alleviated by the blockade of pyroptosis using NLRP3 inhibitor MCC950. Further study demonstrated that RB induced pyroptosis in a caspase-1-dependent manner, as evidenced by the finding that VX765 effectively reversed the effects of RB on A549 and H520 cells. We also found that RB could trigger caspase-1-dependent pyroptosis through ROS-mediated NF-κB suppression. In summary, our findings provide a potential antitumor agent and a novel insight into the mechanism of RB treatment of NSCLC.