Luteolin suppresses gastric cancer progression by reversing epithelial-mesenchymal transition via suppression of the Notch signaling pathway
- Ming-de Zang†1,
- Lei Hu†1,
- Zhi-yuan Fan†1,
- He-xiao Wang1,
- Zheng-lun Zhu1,
- Shu Cao1,
- Xiong-yan Wu1,
- Jian-fang Li1,
- Li-ping Su1,
- Chen Li1,
- Zheng-gang Zhu1,
- Min Yan1 and
- Bing-ya Liu1Email author
© The Author(s) 2017
Received: 24 October 2016
Accepted: 17 February 2017
Published: 27 February 2017
Gastric cancer (GC) is one of the most malignant tumors and the second leading cause of cancer-related deaths in the world. Luteolin, a flavonoid present in many fruits and green plants, suppresses cancer progression. The effects of luteolin on GC cells and their underlying mechanisms remain unclear.
Effects of luteolin on cell proliferation, migration, invasion, and apoptosis were examined in vitro and in vivo by cell counting kit-8 (CCK-8), transwell assays, and flow cytometry, respectively. Real-time reverse transcription polymerase chain reaction (RT-PCR) and Western blots were performed to evaluate Notch1 signaling and activation of epithelial-mesenchymal transition (EMT) in GC cells treated with or without luteolin. Immunohistochemistry was performed to examine proliferation and Notch1 expression in xenograft tumors.
Luteolin significantly inhibited cell proliferation, invasion, and migration in a dose-dependent and time-dependent manner and promoted cell apoptosis. Luteolin reversed EMT by shrinking the cytoskeleton and by inducing the expression of epithelial biomarker E-cadherin and downregulating the mesenchymal biomarkers N-cadherin, vimentin and Snail. Furthermore, Notch1 signaling was inhibited by luteolin, and downregulation of Notch1 had similar effects as luteolin treatment on cell proliferation, migration, and apoptosis. In addition, luteolin suppressed tumor growth in vivo. A higher expression of Notch1 correlated with a poor overall survival and a poor time to first progression. Furthermore, co-immunoprecipitation analysis revealed that activated Notch1 and β-catenin formed a complex and regulated cell proliferation, migration, and invasion.
In this study, GC progression was inhibited by luteolin through suppressing Notch1 signaling and reversing EMT, suggesting that luteolin may serve as an effective anti-tumor drug in GC treatment.
KeywordsGastric cancer Luteolin Notch1 β-Catenin Apoptosis EMT
The incidence of cancer is higher worldwide due to various factors such as smoking, environmental pollution, obesity and aging. Gastric cancer is the fourth most common cancer and the second leading cause of cancer-related deaths in the world . It is the leading cause of tumor-related deaths among males in China . However, effective GC treatment is absent and resistance to chemotherapy is one of its most crucial obstacles, particularly in advanced GC. Due to a lack of validated screening programs, most GC patients are diagnosed at a late stage, leading to a high mortality, in developing countries [3, 4]. Therefore, it is necessary to identify mechanisms underlying GC development as well as design novel drugs for its treatment. Artemisinin, which is isolated from a Chinese herb, suppresses tumor development by causing cell cycle arrest and inducing apoptosis in cancer cells [5, 6]. Luteolin is a flavonoid present in many fruits and green plants, and has the ability to suppress cancer progression [7, 8], which indicates that it may be used as a drug for the treatment of tumors.
Notch signaling is implicated in a majority of cancers for promoting the malignant phenotype by inducing cell proliferation, metastasis, drug resistance, and inhibiting apoptosis [9–12]. Ligand binding to Notch, which is a single-pass transmembrane receptor, leads to its cleavage and release of the Notch intracellular domain (NICD), which translocates to the nucleus and interacts with transcription factor RBPJ to regulate cellular functions [13–15]. The Wnt/β-catenin pathway is conserved across species  and regulates tissue development in embryos and tissue maintenance in adults. Aberrant activation of Wnt/β-catenin promotes the progression of a variety of cancers due to uncontrolled cell proliferation and growth [17, 18]. There is a crosstalk between the Notch and Wnt/β-catenin signaling pathways in many cell types for regulating cell proliferation and migration during development [19, 20]. However, this crosstalk may cause synergistic or antagonistic effects depending on the context [21, 22], and its status in GC remains unclear.
Epithelial-mesenchymal transition (EMT) is not only a physiological process but also a pathological process that regulates cell phenotype and functions during embryogenesis as well as tumor development [23–25]. Morphological changes due to EMT and effects of the tumor microenvironment cause resistance to therapy in many cancers through a number of signaling pathways [26–28]. Notch signaling-induced EMT is a key factor implicated in tumor metastasis [29–31]. Therefore, we addressed the relationship between Notch and EMT in GC progression.
In order to identify the mechanisms underlying GC development as well as effective treatment methods, we studied the therapeutic effect of luteolin on GC and its potential molecular mechanisms of action.
Luteolin inhibits the proliferation and colony formation ability of GC cells
Luteolin promotes apoptosis in GC cells
Luteolin inhibits invasion and migration of GC cells
Luteolin reverses EMT and suppresses Notch1 signaling in GC cells
Luteolin suppresses GC progression via decreasing Notch1 expression
Luteolin suppresses tumor growth in vivo
Although tumor resection and chemotherapy contribute significantly to GC treatment, drug resistance and genetic variation reduce their efficacy . A number of compounds obtained from Chinese herbs possess anti-tumor activities, such as those from Taurine , Dioscorea bulbifera , and Matrine [35, 36]. Although it has been reported that luteolin exerts a marked antitumor effect in cMet-overexpressing patient-derived tumor xenograft models of gastric cancer, luteolin by which way regulating cancer metastasis remains unclear . Luteolin is a flavonoid extracted from Chinese herbs, and in this study, we examined its effects on metastasis in GC and the underlying mechanisms.
Uncontrolled growth of cells is in the hallmark of tumor progression . Hence, we first examined the effects of luteolin on cell proliferation and colony formation in GC, and observed that proliferation of GC cells was significantly inhibited by luteolin in a time- and dose-dependent manner. Next, we found that apoptosis was elevated by luteolin by inhibiting Akt signaling, which itself is implicated in cancer metastasis, proliferation, and apoptosis [39, 40]. We also observed that the cytoskeleton of GC cells shrank upon treatment with luteolin. F-actin staining was used to confirm the changes in the cytoskeleton of GC cells treated with luteolin, and the results suggest that luteolin might inhibit GC motility by reversing EMT in a manner similar to that observed in pancreatic and ovarian cancers . As expected, upon luteolin treatment, the epithelial biomarker E-cadherin was increased. In the contrast, the mesenchymal biomarkers N-cadherin, vimentin, and Snail were reduced in a dose-dependent manner.
Notch signaling was activated during EMT in cardiac development, colorectal cancer, and hepatic carcinoma progression [29, 41]. Notch signaling is predicted to be the potential target of luteolin by our endogenous tumor network model and regulation of Notch signaling plays a key role in tumor progression [42, 43]. Hence, we evaluated the effect of luteolin on Notch signaling, and luteolin treatment indeed inhibited Notch signaling. Silencing of Notch1 inhibited proliferation and migration, induced apoptosis, and reversed EMT of GC cells, which was consistent with the effects observed upon luteolin treatment. Furthermore, overexpression of Notch1 partially recovered EMT and cell migration, illustrating that luteolin suppressed GC progression by inhibiting Notch signaling.
The Notch and Wnt/β-catenin signaling pathways interact in many cell types in synergistic or antagonistic ways, depending on the context [20, 22]; however, it has not been studied in GC. The interaction between β-catenin and NICD was observed in GC cells as well as in leukemia and intestinal stem cells [13, 15], suggesting that they might form a complex to promote GC progression by inducing EMT, elevating proliferation, and inhibiting apoptosis [16, 44]. Luteolin treatment inhibited crosstalk between β-catenin and NICD by decreasing their expression. In an in vivo assay, luteolin suppressed tumor growth by inhibiting proliferation and inducing apoptosis. Luteolin treatment also significantly reduced Notch1 expression. Moreover, higher Notch1 expression was correlated with a poor OS and a poor time to FP . We propose a model for the role of Notch1 and β-catenin in GC progression (Fig. 7). NICD interacts with β-catenin to regulate target genes, causing induction of cell proliferation and metastasis and inhibition of apoptosis. Luteolin inhibits Notch1 and β-catenin expression, thus exhibiting an anti-tumorigenic effect in GC.
In this study, we found that luteolin significantly suppressed GC progression by inhibiting cell proliferation, migration, and invasion; inducing apoptosis; and reversing the EMT. Meanwhile, the Akt, β-catenin, and Notch signaling pathways were inhibited by luteolin. Furthermore, the effects of downregulation of Notch1 were similar to those observed upon treatment of GC cells with luteolin. In addition, higher Notch1 expression correlated with a poor OS. Luteolin suppressed tumor growth and Notch1 signaling in vivo. Therefore, luteolin may be an effective drug for GC treatment.
Cell culture and materials
The human GC cell lines NCI-N87 and MKN28 were maintained in our lab, and Hs-746T was purchased from American Type Culture Collection. Cells were cultured at 37 °C in 5% CO2 and saturation humidity in RPMI-1640 medium with 10% fetal bovine serum containing penicillin and streptomycin. Luteolin was purchased from Aladdin Industrial Corporation (Shanghai). The molecular weight is 286.24 g/mol and the purity is greater than 98% (HPLC).
Cell proliferation and colony formation assays
Cell proliferation was monitored by Cell Counting Kit-8 (CCK-8). In brief, GC cells were suspended in medium with or without luteolin treatment and then plated in 96-well plate at the concentration 2000 cells/well. Cell proliferation was measured every 24 h for 5 days after adding CCK-8 reagent 2 h at the absorbance 450 nm using Epoch Microplate Spectrophotometer (Bio Tek). For colony formation, Hs-746T and MKN28 cells were plated in 6-well plates at the concentration 1000 cells/well. The experimental groups were treated with luteolin for 24 h groups and then replaced for fresh medium. After 10–14 days, the plates were stained with 1% crystal violet.
Hs-746T and MKN28 cells were plated in 6-well plates treated with or without luteolin for 24 h. Then cells were collected and examined by apoptosis detection kit (BD Pharmingen). Briefly, 3 μl annexin V-FITC and 5 μl propidium iodide were added into cells successively for 15 min. And then GC cells were monitored by flow cytometry. Right upper quadrant represents percentage of late apoptosis. Right lower quadrant represents percentage of early apoptosis.
Migration and invasion assays
A number of 1 × 105 GC cells were suspended in serum-free medium with or without Matrigel (BD Bioscience, CA, USA) in upper chambers (Corning Costar, NY, USA) and luteolin or PBS were added into 24-well plates. Next, GC cells were fixed by 10% formalin and stained by 0.5% crystal violet after 24 h. Finally, GC cells that passed through membrane were photographed and counted.
Cells were plated into 8-well glass (Merck Millipore) for overnight and then fixed, permeated and blocked according to protocols. We next stained the cells with β-catenin antibody (1:100; Cell Signaling Technology, CST), followed by incubation with fluorescent secondary antibody for 1 h at room temperature. The nuclei were stained with DAPI. And to visualize the cytoskeleton of GC cells, rhodamine phalloidin (1:20; CST) was used. Slides were analyzed and imaged on a fluorescence microscope.
Vector construction and transfection
Notch1 shRNA, negative control, pCMV-Notch1 (H3176 pLenti-CMV-MCS-HA-3Flag-P2A-EGFPT2A-Puro), and control vectors were purchased from Oobio Corporation (Shanghai, China) and the vectors carry puromycin-resistance function. siRNA sequences of Notch1 or negative control (NC) were as follows: Notch1, GCAACAGCTCCTTCCACTT; NC, TTCTCCGAACGTGTCACGT. Lip2000 (Invitrogen, Carlsbad, USA) was used to transfect vectors into GC cells and then transfected cells were selected by treatment with puromycin. The effects of shRNA and pCMV-Notch1 were confirmed at protein level using western blot assay.
Western blot assay
The method was consist with the previous . In brief, proteins of cells were separated by SDS-PAGE and then were transferred into PVDF membranes. Primary antibody (1:1000 dilutions) AKT, p-AKT, NICD, β-catenin, E-cadherin, N-cadherin, Snail and vimentin were purchased from cell signaling technology (CST, USA). Notch-1, Hes-1, Hey-1, Cyclin-D1 (1:1000 dilutions) and GAPDH (1:10000 dilutions) were purchased from Proteintech. After incubation with primary antibody, secondary antibody followed. Finally, the results were visualized by Tanon system.
Cells were lysed at 4 °C using RIPA and the followed procures were based on Co-IP kit (Pierce, Rockford, USA) according to manufacturer’s instructions. In brief, a total of 300 μg proteins were incubated overnight with specific primary antibodies β-catenin and NICD at 4 °C. Immune complexes were precipitated with protein A/G Sepharose beads and next were examined by western blot.
RNA was extracted from GC cells treated with luteolin or without, and then RNA was reversed to cDNA. Primers for Notch-1, forward GCTTGTGGTAGCAAGGAAGC (20b), reverse CCACATTCAAGTGGCTGATG (20b); Hes-1, forward ACACGACACCGGATAAACCAA (21b), reverse CGAGTGCGCACCTCGGTA (18b); Cyclin-D1, forward GGGTGGGTTGGAAATGAACT (20b), reverse CTTCCTCTCCAAAATGCCAG (20b). RT-PCR was performed using SYBR-green according to manufacturer’s instructions.
In vivo experiment and immunohistochemistry
BALB/c male nude mice (Institute of Zoology, China Academy of Sciences) were used to evaluate the role of luteolin in tumor growth in vivo. Nude mice received humane care and the study protocols were carried out according to a protocol approved by the Institutional Animal Care and Use Committee (IACUC) at Shanghai Jiao Tong University, Shanghai, China. Tumor nodules were measured every week, and was calculated using formula: tumorous volume = (width2 × length)/2. Mice were killed at 4 weeks after injection and then tumors were weighed and fixed for immunohistochemistry staining (IHC).
For IHC, sections staining was performed according to the DAKO protocol, using primary antibody (1:200 dilutions) Notch1, β-catenin and Ki-67. The tunel assay was performed using In situ cell death detection kit (Roche). Blue represents nucleus, green represents death cells.
Differences between experimental groups were assessed by the Student’s t test or one-way ANOVA. Student’s t test was used to examine the statistical differences between the two groups. The significant inhibited effect on cell growth by luteolin was observed at 4th and 5th day after luteolin treatment. The results of 4th and 5th day were compared to that in their control groups using the Student’s t test in Fig. 1a, as well as in Fig. 1b. Survival was analyzed with the Kaplan–Meyer method comparing survival curves by log-rank test. Data are shown as mean ± SD. A two-tailed value of P < 0.05 was considered statistically significant. Statistical analyses were performed using IBM SPSS 19.0 software (SPSS Inc).
Notch intracellular domain
cell counting kit 8
real-time reverse transcription polymerase chain reaction
MDZ and BYL designed this study, MDZ, LH, ZYF performed the qRT-PCR and western blot assays, established xenograft tumor, carried out the immunohistochemistry assay, and drafted the manuscript. HXW, SC, ZLZ, CL, MY and ZGZ participated in the design of the study and analyzed the data. XYW, JFL and LPS helped to acquire experimental data. CS helped to draft the manuscript. All authors read and approved the final manuscript.
The authors declare that they have no competing interests.
Availability of data and materials
The datasets during and/or analysed during the current study available from the corresponding author on reasonable request.
Ethics approval and consent to participate
The study on animals was approved by the Ethics Committee of Ruijin hospital, Shanghai Jiao Tong University School of Medicine. Mice were received humane care and the study protocols were carried out according to a protocol approved by the Institutional Animal Care and Use Committee (IACUC) at Shanghai Jiao Tong University, Shanghai, China.
This study was supported by grants from National Natural Science foundation of China (No. 91529302, No. 81572798, and No. 81272749), Key Projects in the National Science & Technology Pillar Program of China (No. 2014BAI09B03).
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- Van Cutsem E, Sagaert X, Topal B, Haustermans K, Prenen H. Gastric cancer. Lancet. 2016.
- Chen W, Zheng R, Baade PD, Zhang S, Zeng H, Bray F, et al. Cancer statistics in China, 2015. CA Cancer J Clin. 2016;66:115–32.View ArticlePubMedGoogle Scholar
- Riquelme I, Letelier P, Riffo-Campos AL, Brebi P, Roa JC. Emerging role of miRNAs in the drug resistance of gastric cancer. Int J Mol Sci. 2016;17:424.View ArticlePubMedPubMed CentralGoogle Scholar
- Catalano V, Labianca R, Beretta GD, Gatta G, de Braud F, Van Cutsem E. Gastric cancer. Crit Rev Oncol Hematol. 2009;71:127–64.View ArticlePubMedGoogle Scholar
- Chen G, Gong R, Shi X, Yang D, Zhang G, Lu A, et al. Halofuginone and artemisinin synergistically arrest cancer cells at the G1/G0 phase by upregulating p21Cip1 and p27Kip1. Oncotarget. 2016;7:50302.PubMedPubMed CentralGoogle Scholar
- Mondal A, Chatterji U. Artemisinin represses telomerase subunits and induces apoptosis in HPV-39 infected human cervical cancer cells. J Cell Biochem. 2015;116:1968–81.View ArticlePubMedGoogle Scholar
- Dia VP, Pangloli P. Epithelial-to-mesenchymal transition in paclitaxel-resistant ovarian cancer cells is downregulated by luteolin. J Cell Physiol. 2016;232:391–401.View ArticlePubMedGoogle Scholar
- Lin CH, Chang CY, Lee KR, Lin HJ, Chen TH, Wan L. Flavones inhibit breast cancer proliferation through the Akt/FOXO3a signaling pathway. BMC Cancer. 2015;15:958.View ArticlePubMedPubMed CentralGoogle Scholar
- Majumder M, Xin X, Liu L, Tutunea-Fatan E, Rodriguez-Torres M, Vincent K, et al. COX-2 induces breast cancer stem cells via EP4/PI3K/AKT/NOTCH/WNT axis. Stem Cells. 2016;34:2290–305.View ArticlePubMedGoogle Scholar
- Lu J, Ye X, Fan F, Xia L, Bhattacharya R, Bellister S, et al. Endothelial cells promote the colorectal cancer stem cell phenotype through a soluble form of Jagged-1. Cancer Cell. 2013;23:171–85.View ArticlePubMedPubMed CentralGoogle Scholar
- Kwon OJ, Zhang L, Wang J, Su Q, Feng Q, Zhang XH, et al. Notch promotes tumor metastasis in a prostate-specific Pten-null mouse model. J Clin Invest. 2016;126:2626–41.View ArticlePubMedPubMed CentralGoogle Scholar
- Nguyen D, Rubinstein L, Takebe N, Miele L, Tomaszewski JE, Ivy P, et al. Notch1 phenotype and clinical stage progression in non-small cell lung cancer. J Hematol Oncol. 2015;8:9.View ArticlePubMedPubMed CentralGoogle Scholar
- Lopez-Arribillaga E, Rodilla V, Pellegrinet L, Guiu J, Iglesias M, Roman AC, et al. Bmi1 regulates murine intestinal stem cell proliferation and self-renewal downstream of Notch. Development. 2015;142:41–50.View ArticlePubMedGoogle Scholar
- Yamamizu K, Matsunaga T, Uosaki H, Fukushima H, Katayama S, Hiraoka-Kanie M, et al. Convergence of Notch and beta-catenin signaling induces arterial fate in vascular progenitors. J Cell Biol. 2010;189:325–38.View ArticlePubMedPubMed CentralGoogle Scholar
- Gekas C, D’Altri T, Aligue R, Gonzalez J, Espinosa L, Bigas A. β-Catenin is required for T-cell leukemia initiation and MYC transcription downstream of Notch1. Leukemia 2016.
- Giakoustidis A, Giakoustidis D, Mudan S, Sklavos A, Williams R. Molecular signalling in hepatocellular carcinoma: role of and crosstalk among WNT/ss-catenin, Sonic Hedgehog, Notch and Dickkopf-1. Can J Gastroenterol Hepatol. 2015;29:209–17.View ArticlePubMedPubMed CentralGoogle Scholar
- Thompson MD, Monga SP. WNT/beta-catenin signaling in liver health and disease. Hepatology. 2007;45:1298–305.View ArticlePubMedGoogle Scholar
- Xu W, Pang K, Zhou ZG, Chen YF, Mo T, Li M, et al. Dickkopf 2 promotes proliferation and invasion via Wnt signaling in prostate cancer. Mol Med Rep. 2016;14:2283–8.PubMedGoogle Scholar
- Hayward P, Kalmar T, Arias AM. Wnt/Notch signalling and information processing during development. Development. 2008;135:411–24.View ArticlePubMedGoogle Scholar
- Kwon C, Qian L, Cheng P, Nigam V, Arnold J, Srivastava D. A regulatory pathway involving Notch1/beta-catenin/Isl1 determines cardiac progenitor cell fate. Nat Cell Biol. 2009;11:951–7.View ArticlePubMedPubMed CentralGoogle Scholar
- Hayward P, Brennan K, Sanders P, Balayo T, DasGupta R, Perrimon N, et al. Notch modulates Wnt signalling by associating with Armadillo/beta-catenin and regulating its transcriptional activity. Development. 2005;132:1819–30.View ArticlePubMedPubMed CentralGoogle Scholar
- Kwon C, Cheng P, King IN, Andersen P, Shenje L, Nigam V, et al. Notch post-translationally regulates beta-catenin protein in stem and progenitor cells. Nat Cell Biol. 2011;13:1244–51.View ArticlePubMedPubMed CentralGoogle Scholar
- Kalluri R, Weinberg RA. The basics of epithelial-mesenchymal transition. J Clin Invest. 2009;119:1420–8.View ArticlePubMedPubMed CentralGoogle Scholar
- Fan Y, Wang YF, Su HF, Fang N, Zou C, Li WF, et al. Decreased expression of the long noncoding RNA LINC00261 indicate poor prognosis in gastric cancer and suppress gastric cancer metastasis by affecting the epithelial-mesenchymal transition. J Hematol Oncol. 2016;9:57.View ArticlePubMedPubMed CentralGoogle Scholar
- Zang M, Zhang B, Zhang Y, Li J, Su L, Zhu Z, et al. CEACAM6 promotes gastric cancer invasion and metastasis by inducing epithelial-mesenchymal transition via PI3K/AKT signaling pathway. PLoS ONE. 2014;9:e112908.View ArticlePubMedPubMed CentralGoogle Scholar
- Vesuna F, Lisok A, Kimble B, Domek J, Kato Y, van der Groep P, et al. Twist contributes to hormone resistance in breast cancer by downregulating estrogen receptor-alpha. Oncogene. 2012;31:3223–34.View ArticlePubMedGoogle Scholar
- Wang Z, Li Y, Kong D, Banerjee S, Ahmad A, Azmi AS, et al. Acquisition of epithelial-mesenchymal transition phenotype of gemcitabine-resistant pancreatic cancer cells is linked with activation of the notch signaling pathway. Cancer Res. 2009;69:2400–7.View ArticlePubMedPubMed CentralGoogle Scholar
- Junttila MR, de Sauvage FJ. Influence of tumour micro-environment heterogeneity on therapeutic response. Nature. 2013;501:346–54.View ArticlePubMedGoogle Scholar
- Fender AW, Nutter JM, Fitzgerald TL, Bertrand FE, Sigounas G. Notch-1 promotes stemness and epithelial to mesenchymal transition in colorectal cancer. J Cell Biochem. 2015;116:2517–27.View ArticlePubMedGoogle Scholar
- Kostina AS, Uspensky Vcapital Ie C, Irtyuga OB, Ignatieva EV, Freylikhman O, Gavriliuk ND, et al. Notch-dependent EMT is attenuated in patients with aortic aneurysm and bicuspid aortic valve. Biochim Biophys Acta. 2016;1862:733–40.View ArticlePubMedGoogle Scholar
- Yuan X, Wu H, Han N, Xu H, Chu Q, Yu S, et al. Notch signaling and EMT in non-small cell lung cancer: biological significance and therapeutic application. J Hematol Oncol. 2014;7:87.View ArticlePubMedPubMed CentralGoogle Scholar
- Zugazagoitia J, Guedes C, Ponce S, Ferrer I, Molina-Pinelo S, Paz-Ares L. Current challenges in cancer treatment. Clin Ther. 2016;38:1551–66.View ArticlePubMedGoogle Scholar
- Zhang X, Lu H, Wang Y, Liu C, Zhu W, Zheng S, et al. Taurine induces the apoptosis of breast cancer cells by regulating apoptosis-related proteins of mitochondria. Int J Mol Med. 2015;35:218–26.PubMedGoogle Scholar
- Cui H, Li T, Wang L, Su Y, Xian CJ. Dioscorea bulbifera polysaccharide and cyclophosphamide combination enhances anti-cervical cancer effect and attenuates immunosuppression and oxidative stress in mice. Sci Rep. 2016;5:19185.View ArticlePubMedPubMed CentralGoogle Scholar
- Qian L, Liu Y, Xu Y, Ji W, Wu Q, Liu Y, et al. Matrine derivative WM130 inhibits hepatocellular carcinoma by suppressing EGFR/ERK/MMP-2 and PTEN/AKT signaling pathways. Cancer Lett. 2015;368:126–34.View ArticlePubMedGoogle Scholar
- Zhou H, Xu M, Gao Y, Deng Z, Cao H, Zhang W, et al. Matrine induces caspase-independent program cell death in hepatocellular carcinoma through bid-mediated nuclear translocation of apoptosis inducing factor. Mol Cancer. 2014;13:59.View ArticlePubMedPubMed CentralGoogle Scholar
- Lu J, Li G, He K, Jiang W, Xu C, Li Z, et al. Luteolin exerts a marked antitumor effect in cMet-overexpressing patient-derived tumor xenograft models of gastric cancer. J Transl Med. 2015;13:42.View ArticlePubMedPubMed CentralGoogle Scholar
- Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell. 2011;144:646–74.View ArticlePubMedGoogle Scholar
- Ha GH, Park JS, Breuer EK. TACC3 promotes epithelial-mesenchymal transition (EMT) through the activation of PI3K/Akt and ERK signaling pathways. Cancer Lett. 2013;332:63–73.View ArticlePubMedGoogle Scholar
- Chen DL, Zeng ZL, Yang J, Ren C, Wang DS, Wu WJ, et al. L1cam promotes tumor progression and metastasis and is an independent unfavorable prognostic factor in gastric cancer. J Hematol Oncol. 2013;6:43.View ArticlePubMedPubMed CentralGoogle Scholar
- Ma Y, Bian J, Zhang F. Inhibition of perillyl alcohol on cell invasion and migration depends on the Notch signaling pathway in hepatoma cells. Mol Cell Biochem. 2016;411:307–15.View ArticlePubMedGoogle Scholar
- Li S, Zhu X, Liu B, Wang G, Ao P. Endogenous molecular network reveals two mechanisms of heterogeneity within gastric cancer. Oncotarget. 2015;6:13607–27.View ArticlePubMedPubMed CentralGoogle Scholar
- Ao P, Galas D, Hood L, Zhu X. Cancer as robust intrinsic state of endogenous molecular-cellular network shaped by evolution. Med Hypotheses. 2008;70:678–84.View ArticlePubMedGoogle Scholar
- Bertrand FE, Angus CW, Partis WJ, Sigounas G. Developmental pathways in colon cancer: crosstalk between WNT, BMP, Hedgehog and Notch. Cell Cycle. 2012;11:4344–51.View ArticlePubMedPubMed CentralGoogle Scholar
- Szasz AM, Lanczky A, Nagy A, Forster S, Hark K, Green JE, et al. Cross-validation of survival associated biomarkers in gastric cancer using transcriptomic data of 1065 patients. Oncotarget. 2016;7:49322–33.PubMedPubMed CentralGoogle Scholar
- Zang M, Zhang Y, Zhang B, Hu L, Li J, Fan Z, et al. CEACAM6 promotes tumor angiogenesis and vasculogenic mimicry in gastric cancer via FAK signaling. Biochim Biophys Acta. 2015;1852:1020–8.View ArticlePubMedGoogle Scholar