Parthenolide enhances the metronomic chemotherapy effect of cyclophosphamide in lung cancer by inhibiting the NF-kB signaling pathway

BACKGROUND Parthenolide(PTL),a sesquiterpene lactone derived from the medicinal herb Chrysanthemum parthenium,exhibits various biological effects by targeting NF-kB,STAT3,and other pathways.It has emerged as a promising adjunct therapy for multiple malignancies.AIM To evaluate the in vitro and in vivo effect of PTL on cyclophosphamide(CTX)metronomic chemotherapy.METHODS The cytotoxicity of PTL and CTX on Lewis lung cancer cells(LLC cells)was assessed by measuring cell activity and apoptosis.The anti-tumor efficiency was evaluated using a tumor xenograft mice model,and the survival of mice and tumor volume were monitored.Additionally,the collected tumor tissues were analyzed for tumor microenvironment indicators and inflammatory factors.RESULTS In vitro,PTL demonstrated a synergistic effect with CTX in inhibiting the growth of LLC cells and promoting apoptosis.In vivo,metronomic chemotherapy com-bined with PTL and CTX improved the survival rate of tumor-bearing mice and reduced tumor growth rate.Furthermore,metronomic chemotherapy combined with PTL and CTX reduced NF-κB activation and improved the tumor immune microenvironment by decreasing tumor angiogenesis,reducing Transforming growth factorβ,andα-SMA positive cells.CONCLUSION PTL is an efficient compound that enhances the metronomic chemotherapy effects of CTX both in vitro and in vivo,suggesting its potential as a supplementary therapeutic strategy in metronomic chemotherapy to improve the chemotherapy effects.

Parthenolide抑制脑缺血后神经炎症并促进纹状体内神经发生

目的:检测Parthenolide是否抑制脑缺血诱发的神经炎症及其分子机制。方法:MCAO大鼠腹腔注射parthenol-ide(500μg/kg),行BrdU、BrdU-DCX、BrdU-Tuj-1、BrdU-MAP-2、BrdU-GFAP免疫组化和Western blot检测缺血侧纹状体组织TNF-α表达水平。结果:脑缺血增加缺血侧纹状体内TNF-α表达水平,Parthenolide抑制TNF-α的表达并促进缺血侧纹状体内新生细胞的增殖。给予Parthenolide在脑缺血后3 d、7 d或28 d增加BrdU+-DCX+,BrdU+-Tuj-1+,and Br-dU+-MAP-2+阳性细胞数,同时减少BrdU+-GFAP+阳性细胞数。结论:Parthenolide抑制脑缺血诱发的神经炎症,促进非神经发生区-纹状体内神经发生。尚需在此模型上进一步阐明其分子作用机制。

菊花活性成分Parthenolide对人鼻咽癌CNE2细胞线粒体功能和半胱天冬酶活性的影响(英文)

背景:鼻咽癌是我国南方重点防治的恶性肿瘤之一,探讨天然草本菊花活性成分在NPC化学预防的作用,对建立现代康复体系具有重要意义。目的:研究菊花活性成分倍半萜烯内酯(SLs)对人鼻咽癌细胞中线粒体功能和半胱天冬酶活化途径的影响。设计:完全随机对照分组设计。地点和对象:由中山大学公共卫生学院预防医学系完成,对象为低分化CNE2细胞株。干预:对parthenolide(PN)进行剂量-反应和时间效应作用观察,采用噻唑蓝(MTT)颜色反应检测细胞线粒体功能,底物荧光光谱法测定细胞内caspase-9和-3活性,蛋白免疫印迹法检测线粒体内细胞色素C释放和caspase-3酶原降解片段;并应用特异性抑制剂进行细胞内cas-pase途径阻断实验。主要观察指标:细胞线粒体功能、细胞内caspase-9和caspase-3活性、线粒体细胞色素C的释放和caspase-3酶原的降解。结果:PN(1～100μmol/L)作用12h和24h后,MTT颜色反应抑制率随剂量显著增高,呈明显剂量依赖性(Pearson'sr=0.7322,0.7703,P<0.05),IC50分别为252.94μmol/L和49.63μmol/L;但caspase-9和-3活性未见升高,未见细胞色素C释放和caspase-3酶原降解片段形成。PN与caspase抑制剂联合作用后,caspase-9和-3活性明显低于对照和单独PN作用(t=9.146,8.280,27.325,27.450,P<0.05)。

Parthenolide protects human lens epithelial cells from oxidative stress-induced apoptosis via inhibition of activation of caspase-3 and caspase-9

The apoptosis of lens epithehal cells has been proposed as the common basis of cataract formation, with oxidative stress as the major cause. This study was performed to investigate the protective effect of the herbal constituent parthenolide against oxidative stress-induced apoptosis of human lens epithelial （HLE） cells and the possible molecular mechanisms involved. HLE cells （SRA01-04） were incubated with 50 μM H2O2 in the absence or presence of different doses of parthenolide （10, 20 and 50 μM）. To study apoptosis, the cells were assessed by morphologic examination and Annexin V-propidium iodide double staining flow cytometry; to investigate the underlying molecular mechanisms, the expression of caspase-3 and caspase-9 were assayed by Western blot and quantitative RT-PCR, and the activities of caspase-3 and caspase-9 were measured by a Chemicon caspase colorimetric activity assay kit. Stimulated with H202 for 18 h, a high fraction of riLE cells underwent apoptosis, while in the presence ofparthenolide of different concentrations, dose-dependent blocking of HLE cell apoptosis was observed. The expression of caspase-3 and caspase-9 induced by H202 in HLE cells was significantly reduced by parthenolide both at the protein and mRNA levels, and the activation ofcaspase-3 and caspase-9 was also suppressed by parthenolide in a dose-dependent manner. In conclusion, parthenolide prevents HLE cells from oxidative stress-induced apoptosis through inhibition of the activation ofcaspase-3 and caspase-9, suggesting a potential protective effect against cataract formation.

NF-kB Inhibitor Parthenolide Promotes Renal Tubules Albumin Uptake in Type 2 Diabetic Nephropathy

Objective Injured tubular reabsorption is highlighted as one of the causes of increased albuminuria in the early stage of diabetic nephropathy;however,the underlying mechanism has not been fiilly elucidated.In this study,we aimed to explore whether reducing inflammation and remodeling the insulin signaling pathway could improve albumin uptake of renal tubules.Methods 8-week-old male db/db mice(n=8),a type 2 diabetic nephropathy model,administered with nuclear factor kappa-B(NF-kB)inhibitor parthenolide(PTN,1 mg/kg)intraperitoneally every other day for 8 weeks,were as the treatment group.Meanwhile,the age-matched male db/m mice(n=S)and db/db mice(n=8)were treated with saline as the control group and type 2 diabetic nephropathy group.When the mice were sacrificed,blood and urine were collected to examine homeostasis model assessment of insulin resistance(HOMA-IR)and urine albumin creatinine ratio,and kidney samples were used to analyze histopathologic changes with periodic acid-Schiff(PAS)staining,NF-kB p65,phosphorylation of AKT(p-AKT),amnionless and cubilin expressions with immunohistochemistry as well as western blot,and the albumin uptake of renal tubules by using immunofluorescence.In addition,HKC cells were divided into the insulin group treated with insulin alone,the TNF-a group treated with insulin and tumor necrosis factor(TNF-a),and the TNF-a+PTN group exposed to PTN,insulin and TNF-a.The levels of albumin uptake and expression levels of NF-kB p65,p-IRS-l/IRS-1,p-AKT/AKT,amnionless and cubilin in HKC cells were measured.Results Compared with the db/db group,the db/db+PTN group demonstrated decreased levels of HOMA-IR(36.83±14.09 vs.31.07+28.05)and urine albumin creatinine ratio(190.3±7.3 vs.143.0±97.6 mg/mmol);however,the differences were not statistically significant(P>0.05).Periodic acid-Schiff staining showed PTN could alleviate the glomerular hypertrophy and reduce the matrix in mesangial areas of db/db mice.The renal expression of NF-kB p65 was increased and p-AKT(s473)decreased in the db/db group compared with the db/m group(P<0.05).PTN significantly reduced the renal expression of NF-kB p65 and ameliorated the decline of p-AKT(s473)compared with the db/db group(P<0.05).Compared with the db/m group,the expression of amnionless and cubilin decreased and albumin uptake in tubules were reduced in the db/db group(P<0.05),and PTN could significantly increase the expression of cubilin(P<0.05),and improve albumin uptake in tubules.Insulin promoted albumin uptake and the expression of amnionless and cubilin in HKC cells(P<0.05).TNF-a stimulated the expression of NF-kB p65,increased p-IRS-1(s307)and reduced p-AKT(s473)in HKC cells(PV 0.05).In theTNF-a+PTN group,the expression of NF-kB p65 declined and p-IRS-1(s307)and p-AKT(s473)were restored,compared with theTNF-a group(P<0.05).The expression of amnionless and cubilin decreased in theTNF-a group(P<0.05),and PTN could significantly increase the expression of cubilin(P<0.05).Conclusions Inflammation caused damage to insulin signaling,which reduced amnionless-cubilin expression and albumin uptake.PTN could reduce inflammation and remodel the impaired insulin signaling pathway,which promoted the expression of cubilin and albumin uptake.Our study can shed light on the role of inflammation in the reduction of albumin uptake of renal tubules in type 2 diabetic nephropathy.

Parthenolide对人脐静脉内皮细胞增殖和凋亡的作用研究

目的观察Parthenolide对体外培养的人脐静脉内皮细胞(HUVEC)的增殖和凋亡的影响。方法胶原酶消化法培养HUVEC,在HUVEC培养基中加入不同浓度的Parthenolide作用不同的时间,以MTT法检测细胞的增殖活性,TUNEL法检测细胞凋亡。结果与对照组相比,1、5μmol/L Parthenolide组对脐静脉内皮细胞增殖活性的抑制作用无明显影响(P>0.05);10、15、20μmol/L Parthenolide组显著抑制HUVEC的增殖活性(均P<0.01),并且呈时间依赖性和剂量依赖性增加。5μmol/L Parthenolide组在6h、12h、24h时,细胞凋亡指数与对照组相比均无统计学差异(均P>0.05),101、5μmol/L Parthenolide组在6h、12h、24h各个时间段的凋亡指数均高于对照组(均P<0.01);且Parthenolide10μmol/L和15μmol/L组在12和24h的凋亡指数均高于6h(P<0.01),但12和24h上述两组相比无统计学差异(P>0.05)。结论一定浓度范围内的Parthenolide对内皮细胞的增殖和凋亡无明显影响,而高浓度时可同时抑制细胞增殖和诱导HUVEC凋亡。

Parthenolide inhibits the proliferation and induces the apoptosis of human uveal melanoma cells

AIM: To explore the effect of parthenolide(PTL) on human uveal melanoma(UM) cells(C918 and SP6.5 cells) and its molecular mechanism. METHODS: Carboxyfluorescein succinimidyl amino ester(CFSE) assays and cell counting kit-8(CCK-8) were performed to detect the cell viability. Flow cytometry was used to analyze cell cycle and apoptosis. Quantitative realtime polymerase chain reaction(qRT-PCR) and Western blot assays were performed to measure proliferation-related and apoptosis-related factors.RESULTS: Firstly, PTL decreased the viability of C918 and SP6.5 cells in a dose-dependent manner, and the effect of PTL on C918 cells was stronger than on SP6.5;however, it did not affect normal cells. Secondly, PTL increased the proportion of cell number at cell cycle G1 phase in C918 cells, and decreased the proportion of cell number at S phase, but the proportion did not change at G2 phase. In addition, PTL induced the apoptosis of C918 cells, and decreased the expressions of Cyclin D1, B-cell lymphoma-2(Bcl-2) and B-cell lymphoma-extra large(Bcl-XL). Also, PTL increased Cyclin inhibition protein 1(P21), Bcl-2-associated X protein(Bax), Cysteinyl aspartate specific proteinas-3(Caspase-3) and Caspase-9 expression. However, the expression of Caspase-8 was not changed. CONCLUSION: PTL inhibites proliferation and induces apoptosis in UM cells by arresting G1 phase and regulating mitochondrial pathway, however, it does not affect normal cells.

Inhibitory Effects of Parthenolide on the Activity of NF-κB in Multiple Myeloma via Targeting TRAF6

This study examined the mechanism of the inhibitory effect of parthenolide（PTL） on the activity of NF-κB in multiple myeloma（MM）. Human multiple myeloma cell line RPMI 8226 cells were treated with or without different concentrations of PTL for various time periods, and then MTT assay was used to detect cell proliferation. Cell cycle and apoptosis were flow cytometrically detected. The level of protein ubiquitination was determined by using immunoprecipitation. Western blotting was employed to measure the level of total protein ubiquitination, the expression of IκB-α in cell plasma and the content of p65 in nucleus. The content of p65 in nucleus before and after PTL treatment was also examined with immunofluorescence. Exposure of RPMI 8226 cells to PTL attenuated the level of ubiquitinated Nemo, increased the expression of IκB-α and reduced the level of p65 in nucleus, finally leading to the decrease of the activity of NF-κB. PTL inhibited cell proliferation, induced apoptosis and blocked cell cycle. Furthermore, the levels of ubiquitinated tumor necrosis factor receptor-associated factor 6（TRAF6） and total proteins were decreased after PTL treatment. By using Autodock software package, we predicted that PTL could bind to TRAF6 directly and tightly. Taken together, our findings suggest that PTL inhibits the activation of NF-κB signaling pathway via directly binding with TRAF6, thereby suppressing MM cell proliferation and inducing apoptosis.

Parthenolide attenuates LPS-induced activation of NF-κB in a time-dependent manner in rat myocardium

Parthenolide （PTN）, a selective nuclear factor kappa B （NF-κB） inhibitor, has been used extensively to inhibit NF-κB activation. The duration of the inhibitory effect of PTN on NF-kB in vivo remains unclear. This study was to determine whether a lipopolysaccharide （LPS） challenge 6, 12 and 24 h after the administration of PTN could activate NF-κB. Rats were devided into five groups. The rats in the PTN, PTN＋LPS and DMSO groups were injected intraperitoneally with PTN or DMSO. After 6, 12 or 24 h, LPS was administered in LPS and PTN＋LPS groups. The expressions of NF-κB p50, IκBα and p-IκBα were inhibited in both PTN and PTN＋LPS group at end of 6 and 12 h and no effects at 24 h. In summary, myocardial NF-κB expression occurs 1 h after the administration of LPS. PTN blocks this effect given at 6 h and no inhibitory effect 24 h after administration in vivo.

Growth Inhibition Effect of Parthenolide on Human Hepatocellular Carcinomacell Line BEL-7402

To study the effects of parthenolide on human hepatocellular carcinoma cell line BEL-7402 and explore its possible mechanism, human hepatocellular carcinoma cell line BEL-7402 was cultured in vitro and treated with parthenolide of different concentrations. Cells without addition of parthenolide were used as control. The growth of inhibition of cells induced by various concentrations was analyzed by using the MTT assay. The morphologic changes of apoptosis was observed under an inversion microscope by Giemsa staining. The expression levels of PCNA albumen were measured by means of immunohistochemical methods. Parthenolide can inhibit the proliferation of BEL-7402 cells in vitro in adose-dependent manner. Apoptosis with nuclear chromatin condensation, fragmentation and cell shrinkage were found by means of inversion microscopy. Formation of apoptotic bodies was observed by Giemsa staining. The immunohistochemical results show that the expression of PCNA has been decreased. Parthenolide can inhibit the tumor growing of BEL-7402 cells, and induce the apoptosis of cells. The mechanisms of those functions may be via inhibiting the expression of PCNA.

Hydrogen peroxide-induced apoptosis of human lens epithelial cells is inhibited by parthenolide

AIM： To explore the effect of parthenolide on hydrogen peroxide（H_2O_2）-induced apoptosis in human lens epithelial（HLE） cells. METHODS： The morphology and number of apoptotic HLE cells were assessed using light microscopy and flow cytometry. Cell viability was tested by MTS assay. In addition, the expression of related proteins was measured by Western blot assay. RESULTS： Apoptosis of HLE cells was induced by 200 μmol/L H_2O_2, and the viability of these cells was similar to the half maximal inhibitory concentration（IC50）, as examined by MTS assay. In addition, cells were treated with either different concentrations（6.25, 12.5, 25 and 50 mol/L） of parthenolide along with 200 μmol/L H_2O_2 or only 50 μmol/L parthenolide or 200 mol/L H_2O_2 for 24 h. Following treatment with higher concentrations of parthenolide（50 μmol/L）, fewer HLE cells underwent H_2O_2-induced apoptosis, and cell viability was increased. Further, Western blot assay showed that the parthenolide treatment reduced the expression of caspase-3 and caspase-9, which are considered core apoptotic proteins, and decreased the levels of phosphorylated nuclear factor-κB（NF-κB）, ERK1/2 [a member of the mitogen-activated protein kinase（MAPK） family], and Akt proteins in HLE cells. CONCLUSION： Parthenolide may suppress H_2O_2-induced apoptosis in HLE cells by interfering with NF-κB, MAPKs, and Akt signaling.

Culture supernatants of breast cancer cell line MDA-MB-231 treated with parthenolide inhibit the proliferation, migration, and lumen formation capacity of human umbilical vein endothelial cells

Background Parthenolide has been tested for anti-tumor activities, such as anti-proliferation and pro-apoptosis in recent studies. However, little is known about its role in the process of tumor angiogenesis. This study aims to investigate the effects and potential mechanisms of parthenolide on the proliferation, migration and lumen formation capacity of human umbilical vein endothelial cells. Methods Different concentrations of parthenolide were applied to the human breast cancer cell line MDA-MB-231 cells. After 24-hour incubation, the culture supernatants were harvested and used to treat human umbilical vein endothelial cells for 24 hours. Then an inverted fluorescence phase contrast microscope was used to evaluate the human umbilical vein endothelial cells. The secretion of vascular endothelial growth factor （VEGF）, interleukin （IL）-8 and matrix metalloproteinases （MMP）-9 in the culture supernatant of the MDA-MB-231 cells was then measured with enzyme-linked immunosorbent assay （ELISA） assays. Results Suppression of proliferation, migration, and the lumen formation capacity of human umbilical vein endothelial cells was observed in the presence of the culture supernatants from the breast cancer cell line treated with different concentrations of parthenolide. Parthenolide decreased the levels of the angiogenic factors MMP-9, VEGF, and IL-8 secreted by the MDA-MB-231 cells. Conclusions Parthenolide may suppress angiogenesis through decreasing angiogenic factors secreted by breast cancer cells to interfere with the proliferation, migration and lumen-like structure formation of endothelial cells, thereby inhibiting tumor growth. It is a promising potential anti-angiogenic drug.

Parthenolide increases the sensitivity of gastric cancer cells to chemotherapy

OBJECTIVE:To investigate the effect of parthenolide(PTL),a sesquiterpene lactone medicinal compound,on the sensitivity of the gastric cancer cell line SGC7901 and the DPP-and ADR-resistant sublines SGC7901/DDP and SGC7901/ADR to cisplatin[diamminedichloroplatinum(Ⅱ),DDP]and adriamycin(ADR)in vitro.METHODS:SGC7901,SGC7901/DDP,and SGC7901/ADR were treated with various concentrations of PTL alone or in combination with DDP or ADR.The effects on cell proliferation,apoptosis,and expression/activity of several proliferation/apoptosis-related proteins[B-cell lymphoma-2(Bcl-2),cyclin D1,nuclear factor-kappa B(NF-κB),and Caspase-8]and drug transporters(P-glycoprotein and multidrug resistance protein-1)were measured using flow cytometry,Western blotting,and in vitro activity assays.RESULTS:Treatment of SGC7901 cells with PTL inhibited cell growth,increased apoptosis,and sensitized the cells to DPP.Mechanistically,PTL treatment resulted in downregulation of NF-κB activity and Bcl-2 expression,and upregulation of Caspase-8 activity.Similarly,PTL co-treatment of SGC7901/DDP and SGC7901/ADR overcame their resistance to DDP and ADR,respectively,with concomitant inhibition of NF-κB,Bcl-2,Cyclin D1,P-glycoprotein,and multidrug resistance protein-1 expression and/or activity.CONCLUSION:PTL treatment decreases drug resistance in SGC7901,SGC7901/DDP,and SGC7901/ADR cells,as reflected by induction of apoptosis,inhibition of proliferation,downregulation of pro-survival and drug resistance pathways,and upregulation of pro-apoptotic pathways.Our results suggest that co-treatment with PTL may thus complement existing therapies for gastric cancer.

Recent advances on the structural modification of parthenolide and its derivatives as anticancer agents

Parthenolide(PTL)is a sesquiterpene lactone derived from medicinal plant feverfew(Tanacetum parthenium).Recent studies have demonstrated that it has multiple pharmacological activities,especially in the treatment of various hematological and solid cancers.The superior anticancer activity of PTL suggests that it has the potential to be a first-line drug.However,due to the limited physical and chemical properties,as well as bioavailability,structural modification strategies are strongly recommended to improve the anticancer activity.This review describes representative PTL derivatives obtained by different modification strategies,which are reported to exert antiproliferative activities superior to the parent compound PTL.Furthermore,we also summarize their basic mechanisms on cancer-related signaling pathways,so as to explain the potential and characteristics of PTL and its derivatives in cancer therapy.

Parthenolide inhibits proliferation of vascular smooth muscle cells through induction of G_0/G_1 phase cell cycle arrest

Objective： This study is to determine the effect of the natural product parthenolide, a sesquiterpene lactone isolated from extracts of the herb Tanacetum parthenium, on the proliferation of vascular smooth muscle cells （VSMCs）. Methods： Rat aortic VSMCs were isolated and cultured in vitro, and treated with different concentrations ofparthenolide （l 0, 20 and 30 μmol/L）. [^3H]thymidine incorporation was used as an index of cell proliferation. Cell cycle progression and distribution were determined by flow cytometric analysis. Furthermore, the expression of several regulatory proteins relevant to VSMC proliferation including IκBα, cyclooxygenase-2 （Cox-2）, p21, and p27 was examined to investigate the potential molecular mechanism. Results： Treatment with parthenolide significantly decreased the [^3H]thymidine incorporation into DNA by 30%-56% relative to control values in a dose-dependent manner （P〈0.05）. Addition of parthenolide also increased cell population at G0/G1 phase by 19.2%-65.7% （P〈0.05） and decreased cell population at S phase by 50.7%-84.8% （P〈0.05）, which is consistent with its stimulatory effects on p21 and p27. In addition, parthenolide also increased IκBα expression and reduced Cox-2 expression in a time-dependent manner. Conclusion： Our results show that parthenolide significantly inhibits the VSMC proliferation by inducing G0/G1 cell cycle arrest. IκBα and Cox-2 are likely involved in such inhibitory effect ofparthenolide on VSMC proliferation. These findings warrant further investigation on potential therapeutic implications ofparthenolide on VSMC proliferation in vivo.

小白菊内酯逆转肥胖诱导的胰岛素抵抗实验研究

目的:探讨小白菊内酯(Parthenolide)对胰岛素抵抗的逆转作用及分子机制。方法:利用肥胖胰岛素抵抗(obob)动物模型,观察小白菊内酯对小鼠空腹血糖,胰岛素水平,系统性细胞因子及组织胰岛素的敏感性的影响。结果:obob小鼠经过小白菊内酯(7.5mg/kg)治疗后:(1)空腹胰岛素的水平下降约30%,HOMA指数也明显下降;(2)胰岛素受体特异色氨酸的磷酸化水平受到明显抑制;(3)循环中细胞因子PAI-1和MCP-1的水平降低,而resistin无明显变化。结论:小白菊内酯可能通过对PAI-1和MCP-1的抑制作用,影响NF-κB的活性,继而达到逆转胰岛素抵抗和改善胰岛素敏感性。

植物衍生物可杀死白血病的先祖细胞

一种小白菊植物属的衍生化合物Parthenolide可破坏急性髓性白血病（AML）细胞，保留下相对未受损害的正常造血细胞；并通过杀死AML先祖细胞和干细胞根治这种恶性疾病。

The Antitumor Activity and Mechanism of MCL3 in G422 Glioblastoma

Objective:Parthenolide(PTL)induces anti-tumor effects via the nuclear factor kappa B(NF-κB)signaling pathway.MCL3,a PTL derivative,is a sesquiterpene lactone synthesized by the rearrangement and subsequent oxidation of PTL.The aim of this study was to elucidate the antitumor activity and mechanism of action of MCL3 in glioblastoma(GBM).Materials and Methods:The effects of MCL3 on G422 cell proliferation,apoptosis,invasion,and angiogenesis in vitro were measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay,flow cytometry,the cell invasion,and tube formation assays.The subcutaneously transplanted G422 xenograft model was used to detect the effect of MCL3 on tumor growth in vivo.Pathological changes were analyzed by immunohistochemical staining.The effects of MCL3 on NF-κB and Stat3 transcriptional activities were examined using a dual-luciferase reporter assay.Protein levels related to the NF-κB/interleukin(IL)-6/Stat3 signaling pathway were determined using western blot analysis.Results:MCL3 inhibited GBM cell proliferation,invasion,and angiogenesis in a concentration-dependent manner.Moreover,MCL3 decreased the transcriptional activities of NF-κB and Stat3.MCL3 suppressed tumor growth in the subcutaneously transplanted G422 xenograft model,while the inhibition rate was 79%in tumor weight at 40.0 mg/kg.MCL3 blocked the NF-κB/IL-6/Stat3 signaling pathway in G422 cells and tumor tissues,resulting in the downregulation of Stat3 target genes related to apoptosis,invasion,etc.,Conclusion:The results show that MCL3 might inhibit G422 GBM growth partly due to the inhibition of the NF-κB/IL-6/Stat3 signaling pathway.