目的 观察田基黄水提物对肝癌细胞HepG2凋亡的影响并探讨其作用机制.方法 人肝癌细胞HepG2经培养后分为4组,其中,对照(A)组不加入田基黄水提物处理,B、C、D组分别加入浓度为0.5、1.0、2.0 mg/ml的田基黄水提物处理24 h.通过倒置显微镜观察各组细胞形态变化,流式细胞术检测各组肝癌细胞早期凋亡情况及线粒体膜电位的改变;利用逆转录聚合酶链反应(RT-PCR)检测各组线粒体上相关基因B细胞淋巴瘤(Bcl)-2和Bcl-2相关X蛋白(Bax)的mRNA表达.结果 B、C、D组可见人肝癌细胞HepG2出现明显的凋亡形态学变化;流式细胞术检测发现,B、C、D组细胞膜内侧的磷脂酰丝氨酸(PS)外翻,提示早期细胞凋亡.与A组相比,B、C、D组人肝癌细胞HepG2凋亡率明显增加,且呈剂量依赖性(P<0.05);线粒体膜电位检测提示,与A组相比,B、C、D组细胞线粒体膜电位改变率明显升高,并呈剂量依赖性(P<0.05);与A组相比,B、C、D组还明显上调了Bax mRNA表达水平,显著下调Bcl-2 mRNA表达水平(P<0.05).结论 田基黄水提物可通过线粒体凋亡通路来诱导肝癌细胞凋亡.

Background: HAUS6 participates in microtubule-dependent microtubule amplification, but its role in malignancies including colorectal cancer (CRC) has not been explored. We therefore assessed the potential oncogenic activities of HAUS6 in CRC. Results: HAUS6 mRNA and protein expression is higher in CRC tissues, and high HAUS6 expression is correlated with shorter overall survival in CRC patients. HAUS6 knockdown in CRC cell lines suppressed cell growth in vitro and in vivo by inhibiting cell viability, survival and arresting cell cycle progression at G0/G1, while HAUS6 over-expression increased cell viability. We showed that these effects are dependent on activation of the p53/p21 signalling pathway by reducing p53 and p21 degradation. Moreover, combination of HAUS6 knockdown and 5-FU treatment further enhanced the suppression of cell proliferation of CRC cells by increasing activation of the p53/p21 pathway. Conclusion: Our study highlights a potential oncogenic role for HAUS6 in CRC. Targeting HAUS6 may be a promising novel prognostic marker and chemotherapeutic target for treating CRC patients.

Objective:Anti-inflammatory effects of ursolic acid(UA)on a dextran sulfate sodium(DSS)-in-duced experimental murine colitis models was investigated to elucidate its possible molecular mechanisms on in-testinal epithelial cells(IEC).Methods:For in vivo study,a total of 15 male BALB/c mice weighing(20-22)g were randomly divided into three groups:normal control group,DSS model group and DSS+UA treatment group;the mouse colitis model was induced by 3％dextran sodium sulfate(DSS)for 8 days;one of the DSS-in-duced groups was pretreated with UA.The body weight and colon length of mice in each group were measured by physical balance and vernier caliper,respectively.The mice in each group were scored according to the clini-cal disease activity index(DAI)score method.HE staining was used to observe the histopathological changes of colon in each group.The changes of serum amyloid protease A(SAA)and IL-6 expression in colon tissue were measured by ELISA.Using IL-6-stimulated differentiated Caco-2 cells as an in vitro inflammatory model of hu-man intestinal epithelium,the effects of UA on the activation of IL-6/signal transducer and activator of transcrip-tion 3(STAT3)signal pathway in IEC were examined by Western blot for STAT3 phosphorylation.Results:1)Compared with the normal control group,the DAI score was increased,the length of the colon was shortened,and the histological damage was obvious in the DSS model group(P＜0.05);administration of UA significantly reduced the severity of DSS-induced murine colitis as assessed by DAI score,colon length,and histology damage of colon(P＜0.05).2)Compared with the normal control group,the SAA level and the IL-6 level of colon tissue in the DSS model group increased significantly.The DSS-induced increases of SAA and colonic IL-6 levels were reversed by UA treatment(P＜0.05).3)Compared with normal IEC,IL-6 stimulation significantly increased the phosphorylation level of STAT3;STAT3 phosphorylation in IEC-treated with IL-6 and UA was significantly in-hibited compared with only IL-6 stimulation(P＜0.05).Conclusion:Our findings implicate that UA ameliorates DSS-induced colonic inflammation by blocking IL-6/STAT3 signaling pathway,and therefore indicate that UA may have clinical potential as a novel targeted therapy for ulcerative colitis.

目的:研究熊果酸（UA）对葡聚糖硫酸钠（DSS）诱导的实验性小鼠结肠炎模型的抗炎作用,旨在阐明其对肠上皮细胞（IEC）作用的可能分子机制。方法:在体内研究中,将体质量为20～22 g的雄性BALB/c小鼠15只采用随机数字表法分为正常对照组、DSS模型组、DSS+熊果酸治疗组。通过3%DSS诱导8 d,制备小鼠结肠炎模型;其中一组DSS诱导的小鼠结肠炎模型采用UA预处理。各实验组的小鼠体质量和结肠长度分别采用物理天平和游标卡尺测定;参照临床疾病活动指数（DAI）评分法对小鼠进行评分。采用HE染色观察各组结肠组织病理学变化;通过ELISA法测定各组小鼠血液中血清淀粉样蛋白酶A和结肠组织中IL-6的水平。以IL-6刺激分化的Caco-2细胞制备体外人肠上皮细胞炎症模型,通过Western blot检测UA对肠上皮细胞IL-6/STAT3信号通路中STAT3磷酸化的影响。结果:（1）与正常对照组比较,DSS模型组的DAI评分增加、结肠长度缩短和组织损伤明显,差异均具有统计学意义（P＜0.05）;与DSS模型组比较,UA显著抑制了DSS诱导的DAI评分增加、结肠长度缩短和组织学损伤,差异均具有统计学意义（P＜0.05）。（2）与正常对照组比较,DSS模型组的血清淀粉酶（SAA）水平和结肠组织中IL-6水平升高明显,差异均具有统计学意义（P＜0.05）;与DSS模型组比较,UA可显著逆转DSS诱导的血清淀粉酶（SAA）水平和结肠IL-6水平的升高,差异均具有统计学意义（P＜0.05）。（3）与正常肠上皮细胞比较,IL-6刺激显著上调STAT3的磷酸化水平;而UA可显著抑制肠上皮细胞中IL-6诱导的STAT3磷酸化水平的增加,差异具有统计学意义（P＜0.05）。结论:熊果酸可通过阻断IL-6/STAT3信号通路改善了DSS诱导的结肠炎,提示熊果酸在靶向治疗溃疡性结肠炎中可能具有临床应用潜力。

Valuable diagnostic and prognostic biomarkers are urgently needed for colorectal cancer (CRC), which is one of the leading causes of mortality worldwide. Previous studies have reported altered expression of a mucin-like protein Fc fragment of IgG binding protein (FCGBP) in various types of cancer, but its potential diagnostic, prognostic and immunological roles in CRC remain to be determined. Therefore, the aim of current study was to investigate the potential roles of FCGBP in CRC. The present study investigated FCGBP mutations and changes in its expression levels using a combination of microarray and public dataset analyses, as well as immunohistochemistry. The results demonstrated a 10.5% mutation frequency in the FCGBP coding sequence in CRC tissues, and identified decreased FCGBP mRNA or protein expression levels in colorectal adenoma and CRC (compared with those in normal colorectal tissues from healthy control subjects), including pathologically advanced CRC (stage III+IV vs. I+II). Survival analysis using the GEPIA and Kaplan-Meier Plotter databases revealed that low FCGBP expression levels were associated with short overall, disease-free, relapse-free and event-free survival times in patients with CRC. Notably, analysis using the online Tumor IMmune Estimation Resource database revealed a positive correlation between FCGBP expression levels and the extent of infiltrating immune cells, such as B cells and dendritic cells. Consistently, the expression levels of most markers (51/57) for various types of immune cells were significantly correlated with FCGBP expression levels in CRC tissues. These findings suggested that FCGBP may serve as a diagnostic and prognostic biomarker, and that FCGBP may be associated with immune infiltration in CRC.

Background: HAUS6 participates in microtubule-dependent microtubule amplification, but its role in malignancies including colorectal cancer (CRC) has not been explored. We therefore assessed the potential oncogenic activities of HAUS6 in CRC. Methods: We investigated HAUS6 expression and its prognostic value in CRC by microarray, analysis of public datasets, quantitative reverse transcription - polymerase chain reaction (qRT-PCR) and immunohistochemistry. Biological functions of HAUS6 were investigated using loss-of-function and gain-of-function assays in vivo and/or in vitro . Effectors downstream of HAUS6 were identified using cDNA microarray, bioinformatics analysis, qRT-PCR and western-blotting in CRC cells. The putative mechanism by which HAUS6 knockdown elevated p53 and p21 expression were assessed by rescue experiment. The combination effects of HAUS6 knockdown and 5-fluorouracil (5-FU) treatment in cultured CRC cells were further evaluated. Results: HAUS6 mRNA and protein expression is higher in CRC tissues, and high HAUS6 expression is correlated with shorter overall survival in CRC patients. HAUS6 knockdown in CRC cell lines suppressed cell growth in vitro and in vivo by inhibiting cell viability, survival and arresting cell cycle progression at G0/G1, while HAUS6 over-expression increased cell viability. We showed that these effects are dependent on activation of the p53/p21 signalling pathway by reducing p53 and p21 degradation. Moreover, HAUS6 knockdown enhanced the effects of 5-FU treatment in CRC cells by increasing activation of the p53/p21 pathway. Conclusions: Our study highlights a potential oncogenic role for HAUS6 in CRC. Targeting HAUS6 may be a promising novel prognostic marker and chemotherapeutic target for treating CRC patients.

目的 探讨八宝丹(BBD)对胃癌细胞转移的影响. 方法 体外培养胃癌细胞AGS和MGC803,予不同剂量(0.25、0.5、0.75 mg/mL) BBD干预处理.采用MTT法检测细胞的活力,划痕实验观察细胞的损伤修复能力,Transwell实验观察细胞的迁移能力,粘附实验观察细胞的粘附能力. 结果 BBD可抑制AGS和MGC803细胞的活力,抑制细胞的损伤修复能力,抑制细胞的迁移能力,抑制细胞的粘附能力,均呈现明显的剂量依赖作用. 结论 BBD对胃癌细胞的迁移能力具有显著抑制作用,提示BBD可抑制胃癌细胞的转移.

目的 观察八宝丹(BBD)对人脐静脉内皮细胞(HUVEC)增殖和迁移能力的影响. 方法 体外培养HUVEC,使用不同浓度BBD进行干预,MTT法检测HUVEC增殖情况,倒置显微镜观察细胞形态、划痕损伤,Transwell实验观察细胞损伤修复及迁移能力. 结果 BBD能显著降低HUVEC的活力,抑制HUVEC增殖,抑制HUVEC的损伤修复和迁移能力,并呈现剂量依赖性. 结论 BBD具有抑制HUVEC增殖扣迁移的作用.

目的 探讨八宝丹(BBD)对人胃癌细胞增殖与凋亡的影响. 方法 体外培养胃癌细胞AGS和MGC-803,采用不同浓度的BBD(0、0.25、0.5、0.75、1mg/mL)进行干预,MTT法检测细胞活力,计算BBD对细胞增殖的抑制率;倒置显微镜观察细胞生长密度;台盼蓝染色法检测细胞数量;细胞集落形成实验检测细胞集落形成能力;Hoechst染色检测细胞凋亡. 结果 BBD可显著抑制胃癌细胞AGS和MGC-803的细胞活力,抑制细胞增殖,使细胞密度下降,减少细胞数量,抑制细胞集落形成能力,诱导细胞凋亡. 结论 BBD可显著抑制胃癌细胞AGS和MGC-803的增殖及诱导细胞凋亡.

Objective: To explore the effect and underlying mechanism of Pien Tze Huang (PZH) inhibiting colorectal cancer cell via regulating Hedgehog signaling pathway in vivo and in vitro. Methods: Human colorectal cancer cells (HT-29) were treated with different concentrations (0, 250, 500, 1 000 μg/mL) of PZH. Cell viability and cell survival were calculated by MTT assay and colony formation assay. RT-PCR was used to determine the expression of PCNA, Bcl-XL, Bax, Survivin and the core genes of Hedgehog signaling pathway, including Shh, Ptch, Smo and Gli1. Mice were given intragastric administration of 234 mg/(kg·d) dose of PZH or saline daily, 5 days a week, and were taken materials on the 16th day in a nude mouse xenograft study. Tumor samples were analysed by immunohistochemstry for Ki-67, Shh, Ptch, Smo, Gli1. Results: HT-29 cells were inhibited by PZH in a dose dependent manner measured by MTT assay, colony formation assay and in vivo nude mouse xenograft study. The induction of apoptosis as along with the inhibition of cell proliferation was confirmed by down-regulating the expression of Ki-67, PCNA, Bcl-XL and Survivin, and up-regu lating Bax expression. The expression of Shh, Ptch, Smo and Gli1 was significantly suppressed by PZH treatment examined by RT-PCR and immunohistochemstry analysis. Conclusion: In vivo and in vitro study, it suggested that promotion of cancer cell apoptosis and inhibition of proliferation via suppression of Hedgehog signaling pathway might be one of the mechanisms by which PZH treats colorectal cancer.

The present study aimed to detect the impact of the ethanol extract of the Livistona chinensis seed (EELC) on angiogenesis in human umbilical vein endothelial cells (HUVECs). A chorioallantoic membrane (CAM) assay was used to detect the anti-angiogenic activity of EELC in vivo. In vitro, the effect of EELC on the proliferation, migration and angiogenesis of HUVECs was determined by an MTT assay, a wound healing assay and a tube formation assay, respectively. The vascular endothelial growth factor (VEGF)-A and VEGF receptor (VEGFR)-2 protein and mRNA level were measured with ELISA and reverse transcription-semi-quantitative polymerase chain reaction. It was observed that EELC significantly decreased the formation of new vessels in the CAM assay. EELC inhibited the proliferation and migration of HUVECs. The extent of tube formation by HUVECs was also reduced by EELC. In addition, EELC treatment reduced the level of VEGF-A and VEGFR-2 mRNA and protein. The results suggest that EELC inhibits tumor angiogenesis through inhibiting the proliferation and migration of HUVECs, and by downregulating VEGF and VEGFR.

To evaluate the effect of bear bile powder (BBP) on angiogenesis, and investigate the underlying molecular mechanisms.

Angiogenesis, which has a critical role in human tumor growth and development, is tightly regulated by the Notch signaling pathway. Total alkaloids are active components of the plant Rubus alceifolius Poir, which is used for the treatment of various types of cancer. A previous study by our group showed that the total alkaloids of Rubus alceifolius Poir (TARAP) induced hepatocellular carcinoma (HCC) cell apoptosis through the activation of the mitochondria-dependent pathway in vitro and in vivo, as well as inhibited angiogenesis in a chick embryo chorioallantoic membrane model. In the present study, to further analyze the specific mechanisms underlying the antitumor activity of TARAP, a HCC xenograft mouse model was used to assess the effect of TARAP on angiogenesis in vivo. TARAP was found to suppress the expression of vascular endothelial growth factor (VEGF) A and VEGF receptor-2 in tumor tissues, which resulted in the inhibition of tumor angiogenesis. In addition, TARAP treatment was observed to inhibit the expression of Notch1, delta-like ligand 4 and jagged 1, which are key mediators of the Notch signaling pathway. The present study identified that the inhibition of tumor angiogenesis through the suppression of the Notch signaling pathway may be one of the mechanisms through which TARAP may be effective in the treatment of cancer.

Metastasis is the leading cause of cancer-related mortality in almost all types of cancers, including colorectal cancer (CRC). Epithelial-mesenchymal transition (EMT) is a critical process during the metastatic cascade. This process may be a potential target for the diagnosis and treatment of CRC. Pien Tze Huang (PZH), a well-known traditional Chinese formula, has been demonstrated to be clinically effective in treating various types of human malignancies, including CRC. Our published data suggest that PZH can induce apoptosis, as well as inhibit cell proliferation and tumor angiogenesis, thus suppressing CRC growth in vitro and in vivo. We evaluated the therapeutic efficacy of PZH against CRC metastasis using a CRC liver metastasis mouse model to further explore the mechanisms underlying the antitumor action of PZH. MTT, migration, and Matrigel invasion assays were used to assess the effect of PZH on cell viability, migration and invasion. We then established an orthotopic liver metastasis model of colon cancer using microsurgical techniques. Mice were intragastrically administered 234 mg/kg/day dose of either PZH or saline for 14 days. The body and tumor weights of the mice were measured after they were sacrificed. Moreover, we examined the effect of PZH inhibition on liver metastasis. Finally, EMT-related proteins and the TGF-β signaling pathway were assessed using immunohistochemical staining (IHS). The present data revealed that PZH significantly inhibited the migration and invasion of CT-26 cells in a dose-dependent manner, which affirmed the inhibitory effect of PZH on CRC cell metastasis. No significant change was observed between the in vivo primary tumor growth and body weight. However, the control group had five cases of liver metastasis (5/6), whereas one case was found in the PZH group (1/6). Thus, PZH exhibited therapeutic efficacy against CRC metastasis without apparent toxicity. The inhibitory effect of PZH on EMT resulted in an increase in E-cadherin expression, as well as a decrease in N-cadherin expression. In addition, PZH significantly inhibited TGF-β, as well as the phosphorylation of Smad2/3 and Smad4 in the tumor tissues, indicating its suppressive action on TGF-β signaling. These molecular effects ultimately resulted in the inhibition of cancer cell EMT and tumor metastasis.

Objective. We evaluated the effects of total alkaloids of Rubus alceifolius Poir (TARAP) on the migration and invasion of hepatocellular carcinoma (HCC) and furthermore investigated the possible molecular mechanisms mediating its anticancer activity. Methods. We implanted nude mice with human HCC HepG2 cells and fed them with vehicle (physiological saline) or 3 g/kg/day dose of TARAP 5 days per week for 21 days. We determined the in vitro effect of TARAP on the migration and invasion of HepG2 cells by transwell assay. We evaluated SHH signaling components' (SHH, PTCH, SMO, and Gli1) expression levels by reverse transcriptase-polymerase chain reaction and immunohistochemistry. Activity of the matrix metalloproteinases (MMPs) in supernatants was analyzed by zymography. The expression of the MMPs and their specific tissue inhibitor (tissue inhibitor of matrix metalloproteinases, TIMP-1, 2) in HCC tissues was detected by immunohistochemistry. Results. We discovered that TARAP inhibited hepatocellular migration and invasion in a dose-dependent manner in vitro. In addition, TARAP decreased the expression of SHH, PTCH, SMO, and Gli1 in HCC mouse tumors at both transcriptional and translational levels. Moreover, TARAP inhibited the activity of MMP2 and MMP9. We found that TARAP reduced the expression of MMP2 and MMP9, as well as the tissue inhibitor of MMPs. Conclusion. Our study showed that TARAP inhibits HCC migration and invasion likely through suppression of the hedgehog pathway. This may, in part, explain its anticancer properties. These results suggest that total alkaloids in Rubus alceifolius may have potential as a novel antimetastasis drug in the treatment of HCC.

The widely-used Chinese medicinal herb Hypericum japonicum, also known as Hypericum japonicum Thunb or Tianjihuang, displays potent anti-carcinogenic effects against liver cancer. However, the molecular mechanism underlying the therapeutic effects of Hypericum japonicum remains to be elucidated. The present study investigated the in vivo efficacy of ethyl acetate extract of Hypericum japonicum (EAEHJ) against tumor growth in an H22 cell-bearing liver cancer mouse model. Treatment with EAEHJ significantly reduced tumor weight, but had no effect on murine body weight. The results of the present study also showed that EAEHJ induced H22 cell apoptosis in vivo. In addition, the anti-carcinogenic effects of EAEHJ were investigated in vitro. The results of the present study demonstrate that both phospholipid asymmetry in the plasma membrane and mitochondrial membrane potential were deregulated in HepG2 human hepatoma cells, following treatment with EAEHJ. Treatment with EAEHJ also increased the ratio of pro-apoptotic B-cell lymphoma 2 (Bcl-2)-associated X protein (Bax) to anti-apoptotic Bcl-2, and activated the caspase-9 signaling pathway. These results suggest that EAEHJ is able to trigger the apoptosis of liver cancer cells via the mitochondria-dependent pathway.

Tumor metastasis, a complex process involving the spread of malignant tumor cells from a primary tumor site to a distant organ, is a major cause of failure of cancer chemotherapy. Epithelial-mesenchymal transition (EMT) is a critical step for the initiation of cancer metastasis. The processes of EMT and metastasis are highly regulated by a double-negative feedback loop consisting of TGF-beta 1/ZEB pathway and miR-200 family, which therefore has become a promising target for cancer chemotherapy. Pien Tze Huang (PZH), a well-known traditional Chinese formula first prescribed in the Ming Dynasty, has been demonstrated to be clinically effective in the treatment of various types of human malignancy including colorectal cancer (CRC). Our published data proposed that PZH was able to induce apoptosis, inhibit cell proliferation and tumor angiogenesis, leading to the suppression of CRC growth in vitro and in vivo. To further elucidate the mode of action of PZH, in the present study we evaluated its effects on the metastatic capacities of human colorectal carcinoma HCT-8 cells and investigated the underlying molecular mechanisms. We found that PZH significantly inhibited the migration and invasion of HCT-8 cells in a dose-dependent manner. In addition, PZH treatment inhibited the expression kof key mediators of TGF-beta 1 signaling, such as TGF-beta 1, Smad2/3 and Smad4. Moreover, PZH treatment suppressed the expression of ZEB1 and ZEB2, two critical target genes of TGF-(31 pathway, leading to a decrease in the expression of mesenchymal marker N-cadherin and an increased expression of epithelial marker E-cadherin. Furthermore, PZH treatment upregulated the expression of miR-200a, miR-200b and miR-200c. Collectively, our findings in this study suggest that PZH can inhibit metastasis of colorectal cancer cells via modulating TGF-beta 1/ZEB/miR-200 signaling network, which might be one of the mechanisms whereby PZH exerts its anticancer function.

OBJECTIVE:To observe the effect of bear bile powder (BBP) on the STAT3 pathway and its downstream target genes of nude mice hepatocellular carcinoma (HCC) xenograft, and to explore its mechanism for treating HCC.METHODS:The subcutaneous xenograft model was established using HepG2 cells. When the subcutaneous transplanted tumor was formed, naked mice were randomly divided into two groups, the BBP group and the control group. Mice in the BBP group were administered with BBP by gastrogavage, once daily for 3 consecutive weeks, while mice in the control group were administered with normal saline by gastrogavage, once daily for 3 consecutive weeks. The body weight and the tumor volume were measured once per week. By the end of medication, the tumor weight was weighed and the tumor inhibition ratio calculated. The apoptosis of the tumor tissue was detected by TdT-mediated dUTP nick end labeling (TUNEL). The expression of Bcl2-associated X protein (Bax), B cell lymphoma/eukemina-2 (Bcl-2), cyclin-dependent protein kinase (CDK4), cyclinD1 were detected by reverse transcription-polymerase chain reaction (RT-PCR). The protein expression levels of signal transducers and transcription activators 3 (p-STAT3), proliferating cell nuclear antigen (PCNA), Bax, Bcl-2, CDK4, and cyclinD1 were determined by immunohistochemistry.RESULTS:BBP could inhibit the tumor volume and tumor weight, showing statistical difference when compared with the control group (P < 0.01). Results of TUNEL showed that BBP could significantly induce the apoptosis of hepatoma carcinoma cells. Results of RT-PCR showed that BBP could up-regulate the expression of Bax and down-regulate mRNA expression of Bcl-2, CDK4, and cyclinD1. Immunohistochemical results showed that BBP could up-regulate the expression of Bax and inhibit the protein expression of p-STAT3, PCNA, Bcl-2, CDK4, and cyclinD1.CONCLUSION:BBP could induce the apoptosis of hepatoma carcinoma cells and inhibit their proliferation by regulating STAT3 pathway.

Livistona chinensis seeds have been used for centuries to clinically treat various types of cancer. Our published data suggest that Livistona chinensis seeds are able to inhibit hepatocellular carcinoma (HCC) growth in vitro and in vivo via promotion of mitochondrial-dependent apoptosis. To further elucidate the molecular mechanisms of its antitumor activity, in the present study, we used an HCC xenograft mouse model to evaluate the effect of an ethanol extract of Livistona chinensis seeds (EELC) on tumor angiogenesis and on the activation of the Notch pathway. Intratumoral microvessel density (MVD) in HCC xenograft mouse tumors was evaluated via immunohistochemical (IHC) staining for CD31. The mRNA and protein expression of vascular endothelial growth factor A (VEGF-A), VEGFR-2, Notch, Dll4 and Jagged1 was evaluated using RT-PCR and IHC, respectively. We found that EELC profoundly reduced MVD in the HCC mouse tumors, demonstrating the in vivo inhibitory effect of EELC on tumor angiogenesis. In addition, EELC treatment reduced the expression of VEGF-A and VEGFR-2 in tumor tissues. Furthermore, EELC treatment inhibited the expression of Notch, Dll4 and Jagged1. Our findings suggest that Livistona chinensis seeds inhibit tumor angiogenesis through suppression of the Notch pathway.