Xanthohumol attenuates tumour cell-mediated breaching of the lymphendothelial barrier and prevents intravasation and metastasis

Health beneficial effects of xanthohumol have been reported, and basic research provided evidence for anti-cancer effects. Furthermore, xanthohumol was shown to inhibit the migration of endothelial cells. Therefore, this study investigated the anti-metastatic potential of xanthohumol. MCF-7 breast cancer spheroids which are placed on lymphendothelial cells (LECs) induce “circular chemorepellent-induced defects” (CCIDs) in the LEC monolayer resembling gates for intravasating tumour bulks at an early step of lymph node colonisation. NF-κB reporter-, EROD-, SELE-, 12(S)-HETE- and adhesion assays were performed to investigate the anti-metastatic properties of xanthohumol. Western blot analyses were used to elucidate the mechanisms inhibiting CCID formation. Xanthohumol inhibited the activity of CYP, SELE and NF-kB and consequently, the formation of CCIDs at low micromolar concentrations. More specifically, xanthohumol affected ICAM-1 expression and adherence of MCF-7 cells to LECs, which is a prerequisite for CCID formation. Furthermore, markers of epithelial-to-mesenchymal transition (EMT) and of cell mobility such as paxillin, MCL2 and S100A4 were suppressed by xanthohumol. Xanthohumol attenuated tumour cell-mediated defects at the lymphendothelial barrier and inhibited EMT-like effects thereby providing a mechanistic explanation for the anti-intravasative/anti-metastatic properties of xanthohumol.     

Inhibition of tumour spheroid-induced prometastatic intravasation gates in the lymph endothelial cell barrier by carbamazepine: drug testing in a 3D model

Metastatic breast cancer is linked to an undesired prognosis. One early and crucial metastatic step is the interaction of cancer emboli with adjacent stroma or endothelial cells, and understanding the mechanisms of this interaction provides the basis to define new targets as well as drugs for therapy and disease management. A three-dimensional (3D) co-culture model allowing the examination of lymphogenic dissemination of breast cancer cells was recently developed which facilitates not only the study of metastatic processes but also the testing of therapeutic concepts. This 3D setting consists of MCF-7 breast cancer cell spheroids (representing a ductal and hormone-dependent subtype) and of hTERT-immortalised lymph endothelial cell (LEC; derived from foreskin) monolayers. Tumour spheroids repel the continuous LEC layer, thereby generating “circular chemorepellent-induced defects” (CCIDs) that are reminiscent to the entry gates through which tumour emboli intravasate lymphatics. We found that the ion channel blocker carbamazepine (which is clinically used to treat epilepsy, schizophrenia and other neurological disorders) inhibited CCID formation significantly. This effect correlated with the inhibition of the activities of NF-κB, which contributes to cell motility, and with the inactivation of the mobility proteins MLC2, MYPT1 and FAK which are necessary for LEC migration. NF-κB activity and cell movement are prerequisites of CCID formation. On the other hand, the expression of the motility protein paxillin and of the NF-κB-dependent adhesion mediator ICAM-1 was unchanged. Also the activity of ALOX12 was unaffected. ALOX12 is the main enzyme synthesising 12(S)-HETE, which then triggers CCID formation. The relevance of the inhibition of CYP1A1, which is also involved in the generation of mid-chain HETEs such as 12(S)-HETE, by carbamazepine remains to be established, because the constitutive level of 12(S)-HETE did not change upon carbamazepine treatment. Nevertheless, the effect of carbamazepine on the inhibition of CCID formation as an early step of breast cancer metastasis was significant and substantial (~30 %) and achieved at concentrations that are found in the plasma of carbamazepine-treated adults (40–60 μM). The fact that carbamazepine is a drug approved by the US Food and Drug Administration facilitates a “from-bench-to-bedside” perspective. Therefore, the here presented data should undergo scrutiny in vivo.     

Heteronemin, a spongean sesterterpene, inhibits TNFα-induced NF-κB activation through proteasome inhibition and induces apoptotic cell death

In this study, we investigated the biological effects of heteronemin, a marine sesterterpene isolated from the sponge Hyrtios sp. on chronic myelogenous leukemia cells. To gain further insight into the molecular mechanisms triggered by this compound, we initially performed DNA microarray profiling and determined which genes respond to heteronemin stimulation in TNFα-treated cells and which genes display an interaction effect between heteronemin and TNFα. Within the differentially regulated genes, we found that heteronemin was affecting cellular processes including cell cycle, apoptosis, mitogen-activated protein kinases (MAPKs) pathway and the nuclear factor κB (NF-κB) signaling cascade.We confirmed in silico experiments regarding NF-κB inhibition by reporter gene analysis, electrophoretic mobility shift analysis and I-κB degradation. In order to assess the underlying molecular mechanisms, we determined that heteronemin inhibits both trypsin and chymotrypsin-like proteasome activity at an IC₅₀ of 0.4 μM. Concomitant to the inhibition of the NF-κB pathway, we also observed a reduction in cellular viability. Heteronemin induces apoptosis as shown by annexin V-FITC/propidium iodide-staining, nuclear morphology analysis, pro-caspase-3, -8 and -9 and poly(ADP-ribose) polymerase (PARP) cleavage as well as truncation of Bid. Altogether, results show that this compound has potential as anti-inflammatory and anti-cancer agent.     

Key Role of Nuclear Factor-κB in the Cellular Pharmacokinetics of Adriamycin in MCF-7/Adr Cells: The Potential Mechanism for Synergy with 20(S)-Ginsenoside Rh2

We have previously demonstrated that ginsenoside 20(S)-Rh2 is a potent ATP-binding cassette (ABC) B1 inhibitor and explored the cellular pharmacokinetic mechanisms for its synergistic effect on the cytotoxicity of Adriamycin. The present studies were conducted to elucidate the key factors that influenced ABCB1 expression, which could further alter Adriamycin cellular pharmacokinetics. Meanwhile, the influence of 20(S)-Rh2 on the above factors was revealed for explaining its synergistic effect from the view of ABCB1 expression. The results indicated that 20(S)-Rh2 inhibited Adriamycin-induced ABCB1 expression in MCF-7/Adr cells. Subsequent analyses indicated that 20(S)-Rh2 markedly inhibited Adriamycin-induced activation of the mitogen-activated protein kinase (MAPK)/nuclear factor (NF)-κB pathway, NF-κB translocation to the nucleus, and NF-κB binding activity. Furthermore, 20(S)-Rh2 repressed the Adriamycin-enhanced ability of NF-κB to bind to the human multidrug resistance (MDR1) promoter, and MAPK/NF-κB inhibitors and NF-κB small interfering RNA reversed the Adriamycin-induced expression of ABCB1. Moreover, the cellular pharmacokinetics of Adriamycin was also significantly altered by inhibiting NF-κB. In conclusion, the MAPK/NF-κB pathway mediates Adriamycin-induced ABCB1 expression and subsequently alters the cellular pharmacokinetics of Adriamycin. It was speculated that 20(S)-Rh2 acted on this pathway to lower Adriamycin-induced ABCB1 expression in MCF-7/Adr cells, which provided mechanism-based support to the development of 20(S)-Rh2 as a MDR reversal agent.     

1'S-1'-acetoxyeugenol acetate: a novel phenylpropanoid from Alpinia conchigera enhances the apoptotic effects of paclitaxel in MCF-7 cells through NF-κB inactivation

In this study, the apoptotic mechanism and combinatorial chemotherapeutic effects of the cytotoxic phenylpropanoid compound 1'S-1'-acetoxyeugenol acetate (AEA), extracted from rhizomes of the Malaysian ethnomedicinal plant Alpinia conchigera Griff. (Zingiberaceae), on MCF-7 human breast cancer cells were investigated for the first time. Data from cytotoxic and apoptotic assays such as live and dead and poly-(ADP-ribose) polymerase cleavage assays indicated that AEA was able to induce apoptosis in MCF-7 cells, but not in normal human mammary epithelial cells. A microarray global gene expression analysis of MCF-7 cells, treated with AEA, suggested that the induction of tumor cell death through apoptosis was modulated through dysregulation of the nuclear factor-kappaB (NF-κB) pathway, as shown by the reduced expression of various κB-regulated gene targets. Consequent to this, western blot analysis of proteins corresponding to the NF-κB pathway indicated that AEA inhibited phosphorylation levels of the inhibitor of κB-kinase complex, resulting in the elimination of apoptotic resistance originating from NF-κB activation. This AEA-based apoptotic modulation was elucidated for the first time in this study, and gave rise to the proposal of an NF-κB model termed the 'Switching/Alternating Model.' In addition to this, AEA was also found to synergistically enhance the proapoptotic effects of paclitaxel, when used in combination with MCF-7 cells, presumably by a chemosensitizing role. Therefore, it was concluded that AEA isolated from the Malaysian tropical ginger (A. conchigera) served as a very promising candidate for further in-vivo development in animal models and in subsequent clinical trials involving patients with breast-related malignancies.     

原白头翁素衍生物诱导人乳腺癌细胞的凋亡机制

目的:探讨原白头翁素衍生物诱导MCF-7人乳腺癌细胞株凋亡的作用机制。方法:采用体外培养方法培养MCF-7人乳腺癌细胞;应用免疫组织化学技术检测实验组和对照组中死亡受体蛋白(Fas),天冬氨酸蛋白水解酶(Caspase)-3、Caspase-8,细胞核转录因子(NF)-κB的表达。结果:对照组中Fas、Caspase-3和Caspase-8蛋白低表达于细胞质或细胞核,呈弱阳性;实验组细胞这些蛋白表达增高,呈强阳性;2组3种蛋白阳性表达率差异有统计学意义(P<0.05)。对照组细胞中NF-κB高表达于细胞浆和细胞核,而实验组NF-κB表达较对照组下降,呈弱表达于细胞质。结论:原白头翁素衍生物通过外源性途径即死亡受体通路完成凋亡的启动和执行,抑制NF-κB信号转导途径可能是促进MCF-7细胞凋亡的原因。     

补骨脂素逆转GST-π介导的多药耐药的研究

目的　探讨补骨脂素在逆转谷胱甘肽s-转移酶π（GST-π）介导的多药耐药中的作用,以及在MCF-7/ADR细胞中的可能机制。方法　研究时间：2015年至2020年。采用CCK-8法检测细胞活力,评价补骨脂素的细胞毒性和多药耐药逆转活性。采用实时聚合酶链反应（RT-PCR）检测GST-π的表达,检测靶基因的变化。采用免疫印迹法检测GST-π蛋白水平。采用免疫荧光法观察核因子κB（NF-κB）的活化情况。组间比较采用独立样本t检验,P<0.05为差异有统计学意义。结果　应用补骨脂素处理后,细胞内阿霉素药物浓度明显升高。与对照组相比,补骨脂素降低了治疗组GST-π在基因和蛋白水平上的表达。NF-κB抑制剂（SN50）可显著抑制乳腺癌MCF-7/ADR细胞中GST-π的表达。结论　NF-κB信号通路可能是GST-π介导的多药耐药发病机制之一。补骨脂素参与了逆转GST-π介导的多药耐药。GST-π介导的耐药机制可能与NF-κB信号通路有关,是NF-κB信号通路下游的关键因子。     

Motorcycle exhaust particles up-regulate expression of vascular adhesion molecule-1 and intercellular adhesion molecule-1 in human umbilical vein endothelial cells

Epidemiological studies have shown that there is a strong correlation between atherosclerosis and ambient air pollution. In this study, we found that motorcycle exhaust particles (MEP) induced adhesion between cells of the human monocytic leukemia cell line (THP-1) and human umbilical vein endothelial cells (HUVECs) in a time-and dose-dependent manner. In addition, MEP treatment induced both mRNA and protein expression of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) in HUVECs. The IκB degradation and p65 nuclear translocation was found in MEP-treated HUVECs, suggested the involvement of nuclear factor-κB (NF-κB). MEP-induced VCAM-1 and ICAM-1 protein expression was inhibited by NF-κB inhibitor BAY 11-7085. Oxidative stress was also involved in the signaling of VCAM-1 and ICAM-1 expression. MEP treatment caused hydrogen peroxide and superoxide formation. Pretreatment with α-tocopherol could inhibit MEP-induced reactive oxygen intermediates generation and suppressed MEP-induced IκB degradation and adhesion molecules expression. Furthermore, the carbon black (CB) nanoparticles with different diameters could induce VCAM-1 and ICAM-1 protein expression; however, polycyclic aromatic hydrocarbons (PAHs) only increased the expression of ICAM-1 but not that of VCAM-1 in HUVECs. In this study, we found that MEPs could induce ICAM-1 and VCAM-1 expression through oxidative stress and NF-κB activation in HUVECs.     

Gossypol decreases tumor necrosis factor-α-induced intercellular adhesion molecule-1 expression via suppression of NF-κB activity

Gossypol is a yellowish polyphenolic compound originally from cotton plant, which has been shown to exert a potential for anti-cancer and anti-inflammatory effects. However, its molecular mechanism is not thoroughly understood on breast cancer cells known to highly express intercellular adhesion molecule-1 (ICAM-1) for their adhesion and metastasis. This study aims to investigate the effect of gossypol on tumor necrosis factor (TNF)-α-stimulated ICAM-1 via nuclear factor-kappa B (NF-κB) activity. Gossypol was shown to inhibit TNF-α-induced ICAM-1 expression and U937 cell adhesion to MDA-MB-231 and MCF-7 cells. Additionally, TNF-α-induced MDA-MB-231 cell invasion was blocked in the presence of gossypol. Chromatin immunoprecipitation analysis demonstrated that gossypol blocks NF-κB binding on the ICAM-1 promoter regions. Additionally, TNF-α-induced NF-κB activation was completely suppressed in the presence of gossypol. Gossypol did not directly suppress the binding of NF-κB to the DNA but rather inhibited the nuclear translocation of p65 and p50 via phosphorylation and degradation of IκB. We also found that gossypol suppresses NF-κB activation induced by a wide variety of agents, including taxol, okadaic acid, and phorbol myristate acetate. Taken together, gossypol effectively inhibited TNF-α-induced ICAM-1 expression via the suppression of NF-κB activation and in vitro adhesion and invasion in human breast cancer cells.     

Mycoepoxydiene, a fungal polyketide inhibits MCF-7 cells through simultaneously targeting p53 and NF-κB pathways

Mycoepoxydiene (MED) is a cytotoxic polyketide that is isolated from the marine fungal strain Diaporthe sp. HLY-1, which is associated with mangroves; however, the mechanism of action of MED remains unknown. Here, we report the molecular mechanisms of apoptosis activation and growth inhibition induced by MED in MCF-7 cells. The present results show that MED induces DNA damage through the production of reactive oxygen species (ROS), which resulted in the phosphorylation of H2AX and the activation of the Ataxia telangiectasia mutated kinase (ATM) and p53 signaling pathways. In addition, MED increases the accumulation of IκBα and enhances the association between IKKγ and Hsp27 via the activation of Hsp27, which eventually resulted in the inhibition of TNF-α-induced NF-κB transactivation. Therefore, we conclude that MED inhibits MCF-7 cells by simultaneously activating p53 to induce apoptosis and suppressing NF-κB to disrupt cell proliferation. Because small molecules having both of these effects are rare, further exploration of MED as an antitumor lead compound is needed.     

Sorafenib inhibits TPA-induced MMP-9 and VEGF expression via suppression of ERK/NF-κB pathway in hepatocellular carcinoma cells

Invasion by hepatocellular carcinoma (HCC) has been reported to occur via the up-regulation of nuclear factor-kappaB (NF-κB). Sorafenib can improve the overall survival in patients with HCC, however, the association of its inhibitory mechanisms with the inactivation of NF-κB remains unclear. Here, Huh7 cell line transfected with NF-κB-luc2 vector was used to study the effects of sorafenib on NF-κB activity, on expressions of matrix metalloproteinase-9 (MMP-9) and vascular endothelial growth factor (VEGF), which were induced by 12-O-tetradecanoylphorbol-13-acetate (TPA). TPA increased the NF-κB activity and the expressions of MMP-9 and VEGF significantly, but its effects were suppressed by sorafenib in a dose-dependent manner. Similar results were found with PD98059, an inhibitor of extracellular signal-regulated kinase (ERK). Furthermore, transfection of Huh7 cell with an inhibitor of kappaB-α mutant vector, led to reduced TPA-induced MMP-9 and VEGF mRNA expressions. Sorafenib inhibits TPA-induced MMP-9 and VEGF expressions via the suppression of ERK/NF-κB pathway in HCC cells.     

SC-41930, a leukotriene B4 receptor antagonist, inhibits 12(S)-hydroxyeicosatetraenoic acid (12(S)-HETE) binding to epidermal cells.

SC-41930, 7-[3-(4-acetyl-3-methoxy-2-propylphenoxy)-propoxyl]-3, 4-dihydro-8-propyl-2H-1-benzopyran-2-carboxylic acid, a potent leukotriene-B4 (LTB4) receptor antagonist, inhibits in vivo 12-hydroxyeicosatetraenoic acid (12-HETE)-induced neutrophil infiltration, suggesting a potential 12-HETE receptor antagonist effect, as well. Since 12-HETE is assumed to have a pathophysiological role in inflammatory skin diseases, and epidermal cells possess high affinity binding sites for 12(S)-HETE, we studied the effect of SC-41930 on 12(S)-HETE binding to the human epidermal cell line, SCL-II. SC-41930 antagonized the 12(S)-HETE binding to SCL-II cells with a Ki of 480 nM. This Ki value is similar to that obtained for the inhibition of LTB4 binding to human neutrophils. Our results show that SC-41930, in addition to its LTB4 receptor antagonist effect, exhibits 12-HETE receptor antagonist effect as well, and therefore may be of benefit in skin diseases with elevated 12-HETE levels.     

参附对脐静脉内皮细胞Toll样受体4/NF-κB途径的影响

目的观察参附注射液对内毒素介导的人脐静脉细胞Toll样受体4/NF-κB途径及细胞间黏附分子、E-选择素表达的影响。方法健康产妇分娩后的新鲜婴儿脐带,应用胶原酶消化,收集内皮细胞进行培养。实验分为对照组、内毒素组、内毒素+参附组。对照组:M199培养基中不加任何物质。内毒素组:M199培养基中加入脂多糖1 mg/L。内毒素+参附组:预先加入参附注射液10 mL/L,30 min后加入脂多糖1 mg/L。干预1h收集细胞,提取核蛋白测定NF-κB活性,干预12 h后收集细胞检测Toll样受体4、细胞间黏附分子1、E-选择素表达水平。结果内毒素组的Toll样受体4、细胞间黏附分子1、E-选择素表达水平、NF-κB活性均显著高于对照组,内毒素+参附组的Toll样受体4、细胞间黏附分子1、E-选择素表达水平、NF-κB活性均明显低于内毒素组。结论参附注射液可以通过抑制NF-κB活化,减弱Toll样受体4/NF-κB途径,抑制了脂多糖介导的黏附分子表达,从而减轻机体的炎症反应。     

Assessing 12(S)-lipoxygenase inhibitory activity using colorectal cancer cells overexpressing the enzyme.

12(S)-Lipoxygenase (LOX) is regarded as a pro-tumorigenic enzyme and as a potential target for therapy and prevention of cancer so that the search for specific 12(S)-LOX inhibitors is part of drug development strategies. To facilitate the identification of specific 12(S)-LOX inhibitors we have created an assay cell line by introducing a12(S)-LOX expression vector into SW480 colorectal cancer cells. When arachidonic acid was supplied in the medium both transiently and stably overexpressing cells produced 12(S)-hydroxytetraenic acid (HETE) originating from the transfected gene at 4-5-fold the amount obtained from control transfectants. 12(S)-HETE production was 1913.7+/-17.2pg/ml and reached a steady state level 24h after addition of arachidonic acid. To demonstrate the models suitability of 12(S)-LOX overexpressing SW480 cells they were used to measure the inhibitory activity of the plant phenols baicalein, kaempferol, quercetin, nordihydroguaretic acid and resveratrol which are known for their chemopreventive as well as LOX-inhibitory activity in different tumour models. All 5 compounds inhibited 12(S)-HETE production at concentrations below those necessary for growth inhibition.     

番茄红素通过ROS/NF-κB调控乳腺癌干细胞干性和化疗敏感性的研究

目的 探讨番茄红素对乳腺癌干细胞(breast cancer stem cell,BCSC)干性和化疗敏感性的影响及其可能的作用机制。方法 体外培养乳腺癌MCF-7细胞,通过悬浮培养和流式细胞术筛选CD44⁺MCF-7干细胞(BCSC)进行后续实验;BCSC分为对照组和番茄红素组(5 μmol·L^-1),给药24 h后,球体形成试验检测BCSC球体形成大小和数量;对照组和番茄红素组BCSC经不同浓度顺铂(0,5,10,20,40,80 μmol·L^-1)处理24 h,CCK8试验检测BCSC细胞增殖,流式细胞术检测细胞凋亡情况;流式细胞术检测对照组和番茄红素组BCSC中活性氧(reactive oxygen species,ROS)水平;BCSC经10 μmol·L^-1活性氧清除剂N-乙酰半胱氨酸(N-acetyl-L-cysteine,NAC)或10 μmol·L^-1 NF-κB抑制剂BAY 11-7085(BAY)处理24 h后,Western blotting检测细胞中p NF-κB和干性相关蛋白LSD1及SOX9的表达水平。结果 CD44⁺细胞中LSD1和SOX9蛋白表达水平显著高于CD44^-细胞;番茄红素明显降低BCSC球体形成数量和大小;番茄红素显著提高顺铂诱导的BCSC细胞凋亡率;番茄红素显著降低BCSC中ROS水平;番茄红素与NAC均降低细胞中p NF-κB蛋白表达水平;番茄红素、NAC和BAY均抑制细胞中LSD1、SOX9蛋白表达水平,且分别经番茄红素、NAC和BAY处理后,BCSC对顺铂敏感性明显升高。结论 番茄红素可能通过ROS/NF-κB信号通路抑制乳腺癌干细胞干性,增强顺铂治疗的敏感性。     

Curcumin Loaded Chitosan-Protamine Nanoparticles Revealed Antitumor Activity Via Suppression of NF-κB,Proinflammatory Cytokines and Bcl-2 Gene Expression in the Breast Cancer Cells

Nano drug delivery has been recently used to enhance the stability and bioavailability of chemotherapeutic agents. In this study, Chitosan/protamine nanocarrier was synthesized and used to encapsulate curcumin (CUR). The physicochemical properties of the empty carrier (CHPNPs) and curcumin-containing carrier (CU-CHPNPs) were characterized by TEM imaging, Zetasizer, and FT-IR spectroscopy. The antitumor activity of the prepared nanoparticles was assessed by determination of cell count, cell viability, the level of NF-κB, IL-6, and TNF-α and Bcl-2 gene expression in breast cancer cells (MCF-7). The results revealed that the obtained CU-CHPNPs had an average hydrodynamic size of 200 nm, zeta potential of +26.66 mv, and showed a drug encapsulation efficiency of 67%, and drug loading capacity of 40.20%. The cell-based assay showed a significant reduction in the cell viability, and NF-κB, TNF-α, and IL-6 levels upon treatment with CU-CHPNPs as compared to free CUR. Finally, the (CU-CHPNPs) downregulated the expression of the Bcl-2 anti-apoptotic gene more effectively than CUR and the CHPNPs comparing with the β Actin housekeeping gene. This study concluded that the nano-encapsulation of CUR significantly enhances its antitumor efficacy via inhibition of NF-κB, IL-6, and TNF-α and downregulation of Bcl-2.     

NF-κB靶向性哑铃形“圈套”寡核苷酸对海马神经元及神经元样细胞中COX-2表达的影响

目的设计合成靶向性NF-κB的哑铃形圈套ODNs,并检测其对神经元样细胞及海马神经元中NF-κB及环氧化酶-2(COX-2)的抑制作用。方法采用NF-κB靶向性哑铃形圈套ODNs转染培养的PC12细胞,用Western blot方法对PC12细胞内NF-κB和COX-2的蛋白表达进行分析;转染培养的海马神经元再经LPS刺激后,用RT-PCR法检测COX-2mRNA。结果经靶向性NF-κB哑铃形圈套ODNs处理的PC12细胞中NF-κB的表达明显降低(P<0.01),同时伴有COX-2表达明显减少(P<0.01);海马神经元中COX-2mR-NA的表达明显降低(P<0.01)。结论靶向性NF-κB的哑铃形圈套ODNs可抑制神经元样细胞及神经细胞中NF-κB和COX-2的表达,且NF-κB对COX-2具有调控作用。     

Tumour-associated macrophages targeted transfection with NF-κB decoy/mannose-modified bubble lipoplexes inhibits tumour growth in tumour-bearing mice

Abstract

Tumour-associated macrophages (TAM) exhibit an M2 phenotype that promotes tumour progression, and conversion of M2 TAM toward a tumouricidal M1 phenotype is a promising anti-cancer therapy. As NF-κB is a key regulator of macrophage polarization, we developed an in vivo TAM-targeting delivery system that combines mannose-modified bubble liposomes/NF-κB decoy complexes (Man-PEG bubble lipoplexes) and ultrasound (US) exposure. We investigated the effects of NF-κB decoy transfection on TAM phenotype in solid tumour-bearing mice. Post-transfection tumour growth and survival rates were also recorded. Th2 cytokine (IL-10) level in TAM was significantly lower by NF-κB decoy transfection using Man-PEG bubble lipoplexes and US exposure, while Th1 cytokine levels (IL-1β, TNF-α and IL-6) were significantly higher when compared with controls. In addition, mRNA levels of vascular endothelial growth factor, matrix metalloproteinase-9 and arginase were significantly lower in TAM post-NF-κB decoy transfection. Importantly, TAM-targeted NF-κB decoy transfection inhibited tumour growth and prolonged survival rates in mice. Therefore, TAM-targeted NF-κB decoy transfection using Man-PEG bubble lipoplexes and US exposure may be an effective approach for anti-cancer therapy based on TAM phenotypic conversion from M2 toward M1.