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OBJECTIVE: To explore genes potentially co-expressed with cyclin E in gastric cancer and discover possible targets for gastric cancer treatment. METHODS: The Cancer Genome Atlas (TCGA) stomach adenocarcinoma sequencing data were used to predict genes co-expressed with cyclin E. Co-expression genes predicted by cBioPortal online analysis with Pearson correlation coefficient ≥0.4 were analyzed by gene ontology (GO) enrichment annotation using the PANTHER online platform (Ver. 7). Interactions between proteins encoded by these genes were analyzed using the STRING online platform (Ver. 10.5) and Cytoscape software (Ver. 3.5.1). Genes displaying a high degree of connection were analyzed by transcription factor enrichment prediction using FunRich software (Ver. 3). The significant transcription factor and cyclin E expression levels and their impact on gastric cancer progression were analyzed by Western blotting and Kaplan-Meier survival curve analysis. RESULTS: After filtering the co-expression gene prediction results, 78 predicted genes that included 73 protein coding genes and 5 non-coding genes with Pearson correlation coefficient ≥0.4 were selected. The expressions of the genes were considered to be correlated with cyclin E expression. Among the 78 genes co-expressed with cyclin E, 19 genes at the central of the regulatory network associated with cyclin E were discovered. Nuclear transcription factor Y subunit alpha (NF-YA) was identified as a significant transcription factor associated with cyclin E co-expressing genes. Analysis of specimen donors' clinical records revealed that high expression of NF-YA tended to be associated with increased cyclin E expression. The expression of both was associated with progression of gastric cancer. Western blotting results showed that compared with normal tissues, NF-YA and cyclin E were highly expressed in tumor tissues (P < 0.001). Survival curve analysis clearly demonstrated relatively poor overall survival of gastric cancer patients with high cyclin E or high NF-YA expression level, compared to patients with low cyclin E or NF-YA expression (P < 0.05). CONCLUSIONS: NF-YA may promote gastric cancer progression by increasing the transcription of cyclin E and other cell cycle regulatory genes. NF-YA might be a potential therapeutically useful prognostic factor for gastric cancer.
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Previous studies have demonstrated that Licochalcone A possesses anti-inflammatory, anticancer, anti-bacterial, anti-malarial and anti-parasitic activities. In the present study the potential anticancer effects of Licochalcone A on MCF-7 cells were investigated. Licochalcone A significantly decreased cell viability and promoted autophagy and apoptosis, as demonstrated by an MTT assay, acridine orange staining and Annexin V-fluorescein isothiocyanate staining, respectively. Western blot analyses demonstrated that Licochalcone A treatment activated the LC3-II signaling pathway while suppressing the phosphoinositide 3-kinase (PI3K)/RAC-α serine-threonine-protein kinase (Akt)/mammalian target of rapamycin (mTOR) signaling pathway. In addition, Licochalcone A significantly increased caspase-3 activity and significantly decreased B-cell lymphoma-2 expression. The results from the present study indicate that Licochalcone A inhibits PI3K/Akt/mTOR activation, and promotes autophagy and apoptosis in MCF-7 cells.
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OBJECTIVES: The objective of this study was to evaluate the association between metformin therapy and the incidence of gastric cancer (GC) in patients with type 2 diabetes mellitus (T2DM). METHODS: We systemically searched the following databases for studies published between the databases' dates of inception and Nov. 2016: PubMed, Embase, the Cochrane Library, the Web of Science, and the China National Knowledge Infrastructure (CNKI). Hazard ratios (HR)and corresponding 95% confidence intervals (CIs) for the association between metformin therapy and the incidence of GC in patients with T2DM were the outcome measures assessed in this study. STATA 12.0 (Stata Corporation, College Station, Texas, USA) was used to conduct the statistical analysis. RESULTS: A total of seven cohort studies including 591,077 patients met all the criteria for inclusion in the analysis. Our data showed that metformin therapy was associated with a significantly lower incidence of GC in patients with T2DM than other types of therapy (HR=0.763, 95% CI: 0.642Ë0.905). Subgroup analysis showed that patients living in Taiwan benefitted more from metformin therapy than patients living in any other region, as metformin significantly decreased the risk of GC in patients living in Taiwan but did not significantly decrease the risk of GC in patients living in other regions (HR=0.514, 95% CI: 0.384-0.688). The results of the present analysis support the idea that metformin facilitates reductions in the risk of T2DM-related GC. CONCLUSIONS: The risk of GC among patients with T2DM is lower in patients receiving metformin therapy than in patients not receiving metformin therapy.
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Cancer stem cells (CSCs) are defined as a rare subpopulation of undifferentiated cells with biological characteristics that include the capacity for self-renewal, differentiation into various lineages, and tumor initiation. To explore the mechanism of CSCs in esophageal squamous cell carcinoma (ESCC), we focused on Leucine-rich repeat containing G protein-coupled receptor 5 (Lgr5), a target gene of the Wnt signaling pathway, which has been identified as a marker of intestinal stem cells and shown to be overexpressed in several human malignancies. Lgr5 expression was significantly correlated with lymph node metastasis, increased depth of invasion, increased tumor size, advanced differentiation, higher AJCC stage and poorer survival. Silencing of Lgr5 expression in the ESCC cell line KYSE450 by small interfering RNA (siRNA) strongly inhibited cell proliferation, migration and invasion ability, the expression of CSCs-related genes and Wnt/ß-catenin signaling. In addition, Lgr5 was highly expressed in ESCC spheroid body cells, which were identified by high expression of CSCs-related genes, and high tumorigenicity in vivo. Taken together, these results demonstrate that Lgr5 activation of Wnt/ß-catenin signaling is a potential mechanism to promote the progression of ESCC and ESCC stem cell renewal, and Lgr5 may be used as a molecular target for the development of treatments for ESCC.
Sujet(s)
Carcinome épidermoïde/métabolisme , Tumeurs de l'oesophage/métabolisme , Régulation de l'expression des gènes tumoraux , Cellules souches tumorales/métabolisme , Récepteurs couplés aux protéines G/génétique , Adulte , Sujet âgé , Sujet âgé de 80 ans ou plus , Animaux , Marqueurs biologiques tumoraux , Carcinome épidermoïde/mortalité , Carcinome épidermoïde/anatomopathologie , Lignée cellulaire tumorale , Mouvement cellulaire , Prolifération cellulaire , Transformation cellulaire néoplasique/génétique , Modèles animaux de maladie humaine , Tumeurs de l'oesophage/mortalité , Tumeurs de l'oesophage/anatomopathologie , Carcinome épidermoïde de l'oesophage , Femelle , Extinction de l'expression des gènes , Hétérogreffes , Humains , Protéines et peptides de signalisation intercellulaire/génétique , Mâle , Souris , Adulte d'âge moyen , Grading des tumeurs , Stadification tumorale , Pronostic , Voie de signalisation WntRÉSUMÉ
Invasion and metastasis is the major cause of tumor recurrence, difficulty for cure and low survival rate. Excavating key transcription factors, which can regulate tumor invasion and metastasis, are crucial to the development of therapeutic strategies for cancers. PU.1 is a master hematopoietic transcription factor and a vital regulator in life. Here, we report that, compared to adjacent non-cancerous tissues, expression of PU.1 mRNA in metastatic hepatocellular carcinoma (HCC), but not primary HCC, was significantly down-regulated. In addition, levels of PU.1 mRNA in metastatic hepatoma cell lines MHCC97L and MHCC97H were much lower than in non-metastatic Hep3B cells. Transwell invasion assays after PU.1 siRNA transfection showed that the invasion of hepatoma cell lines was increased markedly by PU.1 knockdown. Oppositely, overexpression of PU.1 suppressed the invasion of these cells. However, knockdown and overexpression of PU.1 did not influence proliferation. Finally, we tried to explore the potential mechanism of PU.1 suppressing hepatoma cell invasion. ChIP-qPCR analysis showed that PU.1 exhibited a high binding capacity with miR-615-5p promoter sequence. Overexpression of PU.1 caused a dramatic increase of pri-, pre- and mature miR-615-5p, as well as a marked decrease of miR-615-5p target gene IGF2. These data indicate that PU.1 inhibits invasion of human HCC through promoting miR-615-5p and suppressing IGF2. These findings improve our understanding of PU.1 regulatory roles and provided a potential target for metastatic HCC diagnosis and therapy.
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Carcinome hépatocellulaire/secondaire , Mouvement cellulaire , Régulation de l'expression des gènes tumoraux , Facteur de croissance IGF-II/métabolisme , Tumeurs du foie/anatomopathologie , microARN/génétique , Protéines proto-oncogènes/métabolisme , Transactivateurs/métabolisme , Apoptose , Technique de Western , Carcinome hépatocellulaire/génétique , Carcinome hépatocellulaire/métabolisme , Prolifération cellulaire , Humains , Facteur de croissance IGF-II/génétique , Tumeurs du foie/génétique , Tumeurs du foie/métabolisme , Invasion tumorale , Protéines proto-oncogènes/génétique , ARN messager/génétique , Réaction de polymérisation en chaine en temps réel , RT-PCR , Transactivateurs/génétique , Cellules cancéreuses en cultureRÉSUMÉ
Vasculogenic mimicry (VM) refers to the unique ability of highly aggressive tumor cells to mimic the pattern of embryonic vasculogenic networks, and the presence of VM correlates to an increased risk of metastasis and poor clinical outcome of cancers. Several key molecules, including N-cadherin, have been implicated in VM. However, the role of N-cadherin in the formation of VM in esophageal squamous cell carcinoma (ESCC) had not been elucidated. In this study, firstly we aimed to identify VM patterns in ESCC tissues and to explore their clinical significance. VM was present in 12 out of 56 samples, and ESCC with lymph node metastasis had a higher incidence of VM than that without lymph node metastasis. More importantly, VM channels were associated with the expression of N-cadherin in ESCC tissues. In order to further explore the role of N-cadherin in VM formation and invasion and metastasis in ESCC, secondly, we silenced the expression of N-cadherin with small hairpin RNA in ESCC cell line KYSE-70; herein, we showed that KYSE-70 cells with N-cadherin silencing lost not only the capacity to form tube-like structures on collagen (VM) but also the invasion, metastasis and proliferation ability in KYSE-70 cells in vitro. Taken together, antivascular therapies targeting tumor cell VM may be an effective approach to the treatment of patients with highly metastatic ESCC.
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Antigènes CD/métabolisme , Cadhérines/métabolisme , Carcinome épidermoïde/métabolisme , Carcinome épidermoïde/anatomopathologie , Tumeurs de l'oesophage/métabolisme , Tumeurs de l'oesophage/anatomopathologie , Métastase lymphatique/anatomopathologie , Invasion tumorale/anatomopathologie , Adulte , Sujet âgé , Technique de Western , Lignée cellulaire tumorale , Carcinome épidermoïde de l'oesophage , Femelle , Humains , Immunohistochimie , Mâle , Adulte d'âge moyen , RT-PCR , TransfectionRÉSUMÉ
Stathmin, also called oncoprotein 18, is a founding member of the family of microtubule-destabilizing proteins that play a critical role in the regulation of mitosis. At the same time stathmin has been recognized as one of responsible factors in cancer cells. The aim of this study was to assess stathmin status, its correlations with clinicopathological parameters and its role as a progosnostic marker in EC patients. The protein and mRNA levels of stathmin were examined by immunohistochemistry (IHC) and in situ hybridization in 100EC tissues and adjacent noncancerous tissues. mRNA and protein expression of stathmin in three EC cell lines(EC9706, ECa109, EC1 commonly used in research) were also analyzed using immunocytochemistry, western blot and in situ hybridization. The prognostic value of Stathmin expression within the tumor tissues were assessed by Cox regression and Kaplan-Meier analysis. We showed that stathmin expression was significantly higher in EC tissues than in adjacent noncancerous tissues. High stathmin immunostaining score in the EC was positively correlated with tumor differentiation, Tumor invasion, Lymph node metastases, and TNM stage. In addition, we demonstrated that three EC cell lines examined, were constitutively expressing a high level of stathmin. Of those, EC-1 showed the strongest mRNA and protein expression for the stathmin analyzed. Kaplan-Meier analysis showed that significantly longer 5-year survival rate was seen in EC patients with high Stathmin expression, compared to those with low expression of Stathmin expression. Furthermore, multivariate Cox proportional hazard analyses revealed that Stathmin was an independent factors affecting the overall survival probability. In conclusion, our data provide a basis for the concept that stathmin might be associated with EC development and progression.. High levels of Stathmin expression in the tumor tissues may be a good prognostic marker for patients with EC.
Sujet(s)
Marqueurs biologiques tumoraux/génétique , Carcinomes/génétique , Tumeurs de l'oesophage/génétique , ARN messager/analyse , Stathmine/génétique , Adulte , Sujet âgé , Marqueurs biologiques tumoraux/métabolisme , Carcinomes/métabolisme , Carcinomes/anatomopathologie , Carcinome épidermoïde/génétique , Carcinome épidermoïde/métabolisme , Carcinome épidermoïde/anatomopathologie , Lignée cellulaire tumorale , Évolution de la maladie , Tumeurs de l'oesophage/métabolisme , Tumeurs de l'oesophage/anatomopathologie , Femelle , Humains , Immunohistochimie , Mâle , Adulte d'âge moyen , Analyse multifactorielle , Stadification tumorale , Pronostic , Modèles des risques proportionnels , Stathmine/métabolismeRÉSUMÉ
OBJECTIVE: To study the demethylation effect of arsenic trioxide (As2O3) on ERα-negative human breast cancer MDA-MB-435s cells and its possible mechanisms, and to observe its treatment efficacy in combination with tamoxifen (TAM) after ERα re-expression. METHODS: MTT assay was used to examine the inhibitory effect of As2O3 treatment alone or in combination with TAM on cell proliferation. A nude mouse xenograft model was used to further examine the treatment efficacy in vivo. MSP was used to detect the methylation status of ERα gene after treated with As2O3 in MDA-MB-435s cells and the transplanted tumor tissues. RT-PCR was used to detect the mRNA expression of DNMT1 and Erα. Western bolt was used to detect the DNMT1 and ERα protein expression. The diameter of xenograft tumors was measured weekly, and the tumor growth curve was drawn. RESULTS: The level of proliferation of the MDA-MB-435s cells was significantly suppressed after treatment with different concentration of As2O3 alone or As2O3 combined with TAM, and the 4 µmol/L As2O3 + TAM treatment for 72 h showed the highest inhibition rate (62.6%). 1, 2, 4 µmol/L As2O3 had demethylation effect on MDA-MB-435s cells, and the DNMT1 mRNA and protein expression was inhibited and accompanied by ERα mRNA and protein re-expression. The unmethylation specific bands of ERα gene were enhanced after treated by As2O3 alone or As2O3 combined with TAM in the xenograft tumors. The expression of DNMT1 mRNA and protein was inhibited, and accompanied by ERα mRNA and protein re-expression. An significant decrease of volume and weight of the xenograft tumors in the As2O3 treated alone or combined with TAM groups was observed compared with those of the normal saline group or TAM alone group (P < 0.05), and the 4 mg/kg As2O3 + TAM group had the highest inhibition rate of tumor weight (79.5%) and volume (76.4%). CONCLUSIONS: ERα can be re-expressed in ERα-negative breast cancer MDA-MB-435s cells after treated with As2O3 by inhibiting the DNMT1 activity. MDA-MB-435s cells are re-sensitized to endocrine therapy after ERα re-expression. As2O3 combined with TAM may provide a new therapeutic approach for patients with ERα-negative breast cancer in the clinic.
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Antinéoplasiques/pharmacologie , Composés de l'arsenic/pharmacologie , Tumeurs du sein/anatomopathologie , Récepteur alpha des oestrogènes/métabolisme , Oxydes/pharmacologie , Tamoxifène/administration et posologie , Charge tumorale/effets des médicaments et des substances chimiques , Animaux , Antinéoplasiques/administration et posologie , Antinéoplasiques hormonaux/administration et posologie , Protocoles de polychimiothérapie antinéoplasique/pharmacologie , Trioxyde d'arsenic , Composés de l'arsenic/administration et posologie , Tumeurs du sein/génétique , Tumeurs du sein/métabolisme , Lignée cellulaire tumorale , Prolifération cellulaire/effets des médicaments et des substances chimiques , DNA (Cytosine-5-)-methyltransferase 1 , DNA (cytosine-5-)-methyltransferase , Méthylation de l'ADN , Relation dose-effet des médicaments , Récepteur alpha des oestrogènes/génétique , Femelle , Humains , Souris , Souris de lignée BALB C , Souris nude , Oxydes/administration et posologie , ARN messager/métabolisme , Tests d'activité antitumorale sur modèle de xénogreffeRÉSUMÉ
OBJECTIVE: To detect the expression of histone deacetylase 6 (HDAC6) in laryngeal squamous cell carcinoma, and to analyze the effects of downregulation of HDAC6 expression on cell cycle, proliferation and migration of laryngeal squamous cell carcinoma cell line Hep-2 cells, and to explore their possible molecular mechanisms. METHODS: Immunohistochemistry was used to detect the expression of HDAC6 protein in 55 cases of laryngeal squamous cell carcinoma and 20 cases of normal laryngeal mucosa. HDAC6 siRNA and control siRNA were transfected into Hep-2 cells via lipofectamine 2000, and the interfering effect was analyzed using Western blotting. The effects of downregulation of HDAC6 expression on cell cycle, proliferation and migration were determined by cell counting kit-8 (CCK-8), flow cytometry and Boyden chamber, respectively. Finally, Western blotting was used to detect the expressions of cell cycle, proliferation and migration related proteins. RESULTS: There was a high level expression of HDAC6 protein in laryngeal squamous cell carcinoma, and its expression was not related to age and sex of the patients (P > 0.05), but closely associated with the degree of histological differentiation, TNM staging and lymph node metastasis (P < 0.05). HDAC6 siRNA effectively down-regulated the expression of HDAC6 protein in laryngeal squamous cell carcinoma cell line Hep-2 cells, and downregulation of its expression obviously inhibited cell proliferation, arrested cell cycle at G(0)/G(1) phase and decreased cell migration ability in Hep-2 cells. Additionally, the downregulation of HDAC6 protein expression markedly decreased the expressions of cyclin D1, cyclin E, cdk2 and MMP-9 proteins, but increased the expressions of p21 and E-cadherin proteins. CONCLUSIONS: HDAC6 may play a pivotal role in the carcinogenesis and development of laryngeal squamous cell carcinoma. The downregulation of HDAC6 expression-mediated cell proliferation inhibition, cell cycle arrest and decreased cell migration ability may be closely associated with the decrease of cyclin D1, cyclin E, cdk2 and MMP-9 proteins and increase of p21 and E-cadherin proteins.
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Carcinome épidermoïde/anatomopathologie , Cycle cellulaire , Mouvement cellulaire , Prolifération cellulaire , Histone deacetylases/métabolisme , Tumeurs du larynx/anatomopathologie , Adulte , Sujet âgé , Antigènes CD , Cadhérines/métabolisme , Carcinome épidermoïde/génétique , Carcinome épidermoïde/métabolisme , Lignée cellulaire tumorale , Cycline D1/métabolisme , Cycline E/métabolisme , Kinase-2 cycline-dépendante/métabolisme , Régulation négative , Femelle , Histone deacetylase 6 , Histone deacetylases/génétique , Humains , Tumeurs du larynx/génétique , Tumeurs du larynx/métabolisme , Métastase lymphatique , Mâle , Matrix metalloproteinase 9/métabolisme , Adulte d'âge moyen , Stadification tumorale , Protéines oncogènes/métabolisme , Protéines proto-oncogènes p21(ras)/métabolisme , Petit ARN interférent/génétique , TransfectionRÉSUMÉ
OBJECTIVE: To investigate the expression and significance of matrix metalloproteinases (MMP-2, MMP-9) and tissue inhibitor of matrix metalloproteinase (TIMP-2, TIMP-1) in non-melanoma skin cancer (NMSC). METHODS: Thirty six patients with squamous cell carcinoma (SCC) and 32 patients with basal cell carcinoma (BCC), confirmed by pathology, were selected, and 30 cases of normal skin were selected as control. The expression of MMP-2, MMP-9, TIMP-1 and TIMP-2 in all samples were examined by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR). The expression rate, expression intensity and expression level of each factor were recorded. The results were compared between the groups. RESULTS: The expression rates of MMP-2 and MMP-9 in the control group were 30.0% and 36.7%, the expression levels of MMP-2 and MMP-9 in the control group were 57.216 ± 12.785 and 59.318 ± 13.262, all significantly lower than those in the tumor edge and center of the SCC and BCC groups (P < 0.01). The expression rates of TIMP-1 and TIMP-2 in the control group were 96.7% and 100%, their expression levels were 121.738 ± 25.516 and 122.612 ± 25.964, all significantly higher than those in the SCC and BCC groups (P < 0.01). The expression levels of MMP-2 and MMP-9 in the tumor center and edge of SCC group were significantly higher than those in the corresponding parts of the BCC group, while the expression levels of TIMP-1 and TIMP-2 were significantly lower than those in the BCC group (P < 0.01). The expression levels of MMP-2 and MMP-9 in the tumor edge of the SCC and BCC groups were significantly higher than those in the tumor centers (P < 0.01), while the expression levels of TIMP-1and TIMP-2 were significantly lower than those in the tumor centers (P < 0.01). CONCLUSION: MMP-2, MMP-9 and TIMP-2, TIMP-1 may play an important role in the development, progression, invasion and metastasis of non-melanoma skin cancer.
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Matrix metalloproteinase 2/métabolisme , Matrix metalloproteinase 9/métabolisme , Tumeurs cutanées/métabolisme , Inhibiteur tissulaire de métalloprotéinase-1/métabolisme , Inhibiteur tissulaire de métalloprotéinase-2/métabolisme , Sujet âgé , Carcinome basocellulaire/génétique , Carcinome basocellulaire/métabolisme , Carcinome basocellulaire/anatomopathologie , Carcinome épidermoïde/génétique , Carcinome épidermoïde/métabolisme , Carcinome épidermoïde/anatomopathologie , Femelle , Humains , Immunohistochimie , Mâle , Matrix metalloproteinase 2/génétique , Matrix metalloproteinase 9/génétique , Adulte d'âge moyen , ARN messager/métabolisme , Tumeurs cutanées/génétique , Tumeurs cutanées/anatomopathologie , Inhibiteur tissulaire de métalloprotéinase-1/génétique , Inhibiteur tissulaire de métalloprotéinase-2/génétiqueRÉSUMÉ
Curcumin (1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione), is extracted from the plant Curcuma longa. It was recently reported for its anticancer effect on several types of cancer cells in vitro however, the molecular mechanisms of this anticancer effect are not fully understood. In the present study, we evaluated the effects of curcumin on human mammary epithelial carcinoma MCF-7 cells. Cells were treated with curcumin and examined for cell viability by MTT assay. The cells invasion was demonstrated by transwell assay. The binding activity of NF-κB to DNA was examined in nuclear extracts using Trans-AM NF-κB ELISA kit. Western blot was performed to detect the effect of curcumin on the expression of uPA. Our results showed that curcumin dose-dependently inhibited (P < 0.05) the proliferation of MCF-7 cells. Meanwhile, the adhesion and invasion ability of MCF-7 cells were sharply inhibited when treated with different concentrations of curcumin. Curcumin also significantly decreased (P < 0.05) the expression of uPA and NF-κB DNA binding activity, respectively. It is concluded that curcumin inhibits the adhesion and invasion of MCF-7 cells through down-regulating the protein expression of uPA via of NF-κB activation. Accordingly, the therapeutic potential of curcumin for breast cancer deserves further study.
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Tumeurs du sein/traitement médicamenteux , Tumeurs du sein/anatomopathologie , Curcumine/usage thérapeutique , Évolution de la maladie , Facteur de transcription NF-kappa B/métabolisme , Transduction du signal , Activateur du plasminogène de type urokinase/métabolisme , Adhérence cellulaire/effets des médicaments et des substances chimiques , Lignée cellulaire tumorale , Survie cellulaire/effets des médicaments et des substances chimiques , Curcumine/pharmacologie , ADN tumoral/métabolisme , Femelle , Humains , Invasion tumorale , Métastase tumorale , Liaison aux protéines/effets des médicaments et des substances chimiques , Transduction du signal/effets des médicaments et des substances chimiquesRÉSUMÉ
BACKGROUND: The potential application of retinoic acid receptor activators, such as all trans-retinoic acid (ATRA), for treating various cancers have been studied both pre-clinically and clinically. Whether ATRA has an anticancer effect on human esophageal squamous cancer cell (ESCC) is still unknown. We have explored the anticancer effect of ATRA in ESCC, and in this study, the effects of ATRA on levels and patterns of expression of the vascular endothelial growth factor (VEGF) signal transduction pathway in transplantable tumor growth of the human ESCC cell line, EC9706, in nude mice. METHODS: The animal model of the ESCC xenograft was made by subcutaneous implantation of tumor cells into nude mice. Reverse transcription-polymerase chain reaction (RT-PCR), Western blotting and immunohistochemical assays were used to detect the expression of the VEGF signal transduction pathway in ESCC xenograft tissues. RESULTS: Compared to the control group, the tumor inhibition rates in the low dose ATRA, high dose ATRA, and 5-FU groups were 83.21%, 88.32%, 91.02%, respectively. The protein and mRNA levels of VEGF were down-regulated after being treated with ATRA and 5-FU compared to the control group (P < 0.05). The study also revealed that ATRA specifically down-regulated VEGF and the component of the VEGF signal transduction pathway of CD31, CD34, and CD105 (component of the TGF-ß receptor) in ESCC xenograft tissues (P < 0.05). CONCLUSIONS: ATRA can significantly inhibit tumor growth and has anticancer effects on transplantable tumor growth of human ESCC cell line EC9706 in nude mice. These findings indicate that ATRA specifically down regulated VEGF and the components of VEGF signal transduction, which may be an important mechanism responsible for the neoangiogenesis inhibition of ESCC cells.
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Carcinome épidermoïde/traitement médicamenteux , Tumeurs de l'oesophage/traitement médicamenteux , Néovascularisation pathologique/traitement médicamenteux , Trétinoïne/usage thérapeutique , Animaux , Technique de Western , Carcinome épidermoïde/métabolisme , Lignée cellulaire tumorale , Tumeurs de l'oesophage/métabolisme , Humains , Immunohistochimie , Souris , Souris nude , Néovascularisation pathologique/métabolisme , Réaction de polymérisation en chaine en temps réel , Facteur de croissance endothéliale vasculaire de type A/métabolisme , Tests d'activité antitumorale sur modèle de xénogreffeRÉSUMÉ
OBJECTIVE: The aim of this study was to assess the TOP2A RNA expression and the relationship of TOP2A protein expression with metastasis-free interval in breast cancer patients. METHODS: TOP2A expression was analyzed prior to surgery in 86 patients. The level of TOP2A gene amplification was analyzed by fluorescence in situ hybridization (FISH), its RNA expression level with RT-PCR, and their correlation with TOP2A protein expression was assessed by immunohistochemistry (IHC). The correlation between RNA expression level and metastasis-free interval in breast cancer patients was also analyzed. RESULTS: Aberrations (amplification or deletion) of TOP2A copy number was observed in 25.6% (22/86) of the cases. TOP2A protein expression was detected in 66.3% (57/86) of the samples. There was a significant correlation between the TOP2A RNA expression and protein expression (P < 0.001). TOP2A gene expression was significantly associated with the metastasis-free interval in the breast cancer patients (P = 0.001). There was no significant correlation between TOP2A gene amplification and TOP2A protein expression (P = 0.211). CONCLUSIONS: TOP2A RNA level is an objective and reliable prognostic indicator in breast cancer.
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Antigènes néoplasiques/métabolisme , Tumeurs du sein/génétique , Carcinome canalaire du sein/génétique , ADN topoisomérases de type II/métabolisme , Protéines de liaison à l'ADN/métabolisme , Antigènes néoplasiques/génétique , Tumeurs du sein/traitement médicamenteux , Tumeurs du sein/métabolisme , Tumeurs du sein/chirurgie , Carcinome canalaire du sein/traitement médicamenteux , Carcinome canalaire du sein/métabolisme , Carcinome canalaire du sein/chirurgie , Carcinome intracanalaire non infiltrant/traitement médicamenteux , Carcinome intracanalaire non infiltrant/génétique , Carcinome intracanalaire non infiltrant/métabolisme , Carcinome intracanalaire non infiltrant/chirurgie , Carcinome lobulaire/traitement médicamenteux , Carcinome lobulaire/génétique , Carcinome lobulaire/métabolisme , Carcinome lobulaire/chirurgie , Traitement médicamenteux adjuvant , ADN topoisomérases de type II/génétique , Protéines de liaison à l'ADN/génétique , Survie sans rechute , Femelle , Amplification de gène , Délétion de gène , Régulation de l'expression des gènes tumoraux , Humains , Adulte d'âge moyen , Traitement néoadjuvant , Protéines liant le poly-adp-ribose , ARN/métabolisme , Induction de rémissionRÉSUMÉ
BACKGROUND AND OBJECTIVES: Stathmin plays a critical role in the regulation of mitosis and mediates the development of malignant tumors. Here, we investigated the potential role of stathmin in cell cycle and apoptosis in esophageal squamous cell carcinoma (ESCC). METHODS: A stathmin short hairpin RNA (shRNA) plasmid was employed to downregulate stathmin expression in the ESCC cell line EC9706 cells. Cell proliferation was measured by cell counting, MTT, and colony formation assay. Cell migration was measured by Boyden chamber. Western blot was used to analyze the expressions of stathmin, survivin, and apoptosis-related proteins in transfected cells. Cell cycle and apoptosis were determined by flow cytometry and DNA ladder. Oncogenicity assay in nude mice was utilized to analyze phenotypic changes of transfected cells in vivo. RESULTS: After transfection with stathmin shRNA plasmid, stathmin expression markedly decreased in EC9706 cells. Stathmin downregulation significantly inhibited cell proliferation, cell migration in vitro, and tumorigenicity in vivo, meanwhile arrested cell cycle in the G2/M phase and induced cell apoptosis. Further, stathmin downregulation resulted in downregulation of Bcl-2 and survivin proteins, activation of Caspase-3. CONCLUSIONS: These findings demonstrate that stathmin may play an essential role in carcinogenesis of ESCC, which will lay a foundation for target therapy of ESCC.
Sujet(s)
Apoptose/génétique , Carcinome épidermoïde/métabolisme , Régulation négative , Tumeurs de l'oesophage/métabolisme , Stathmine/métabolisme , Animaux , Carcinome épidermoïde/anatomopathologie , Caspase-3/métabolisme , Cycle cellulaire/génétique , Lignée cellulaire tumorale , Mouvement cellulaire , Prolifération cellulaire , Tumeurs de l'oesophage/anatomopathologie , Cytométrie en flux , Humains , Protéines IAP/métabolisme , Mâle , Souris , Souris nude , Phénotype , Plasmides , Protéines proto-oncogènes c-bcl-2/métabolisme , Petit ARN interférent , Survivine , TransfectionRÉSUMÉ
OBJECTIVE: To investigate the inhibitory effect and apoptosis induction on human esophageal carcinoma EC9706 cell by Fufangkushen. METHODS: The experiment of Fufangkushen was designed into three groups including 25.00 µl/ml group, 6.25 µl/ml group and control group in vitro. The method of MTT was used to evaluate the growth inhibition effects. Proliferating cell nuclear antigen (PCNA) was detected by immunohistochemistry (IHC) in vitro. The morphological changes of cells were observed under inverted microscope. FACS was used to analyze the distribution of cell cycle and apoptosis. The expressions of Bcl-2, Fas and caspase-3 in EC9706 cells were detected by Western blotting. The clone formation in plate was used to test the capacity of cell clone formation. Nude mice experiments were conducted to investigate the tumor inhibition of Fufangkushen in vivo. The mice were divided into 3 groups of 200 µl/d treatment, 25 µl/d treatment and saline control. PCNA and Bcl-2 were detected by IHC. And the apoptotic index was detected by terminal transferase dUTP nick end labeling (TUNEL) on xenograft of nude mice. RESULTS: The proliferative capacities of 25.00 µl/ml group were lower than that of the control group at 48, 72, 96 h respectively (all P < 0.01). IHC showed the PCNA expressions, cell clone formation rate were both lower than that of control group (in 25.00 µl/ml treatment group both P < 0.05). Many apoptotic cells could be observed. And the apoptotic rate was higher in 25.00 µl/ml group than that in the control group ((25.2 ± 7.3)% vs (3.4 ± 1.5)%, P < 0.01). After a treatment of Fufangkushen, the activation of caspase-3 and the Fas were higher ((21.3 ± 4.4)% vs (1.8 ± 0.6)%, (30.2 ± 8.3)% vs (5.4 ± 1.6)%, both P < 0.01), the Bcl-2 were lower (P < 0.01) were observed in vitro. Comparing with the saline control group, the tumor weight in 200 µl/d treatment group were lower ((987 ± 386) vs (1935 ± 838) mg, P < 0.01) and the apoptotic index higher ((33.8 ± 8.7)% vs (5.3 ± 1.4)%, P < 0.01). CONCLUSION: Fufangkushen can inhibit the proliferation of EC9706 cells and induce the cellular apoptosis. The mechanism of apoptosis is probably associated with the arrest of cell cycle, the up-regulation of Fas, the down-regulation of Bcl-2 and the activation of caspase-3 in ESCC EC9706 cells.
Sujet(s)
Apoptose/effets des médicaments et des substances chimiques , Carcinome épidermoïde/anatomopathologie , Médicaments issus de plantes chinoises/pharmacologie , Tumeurs de l'oesophage/anatomopathologie , Animaux , Caspase-3/métabolisme , Lignée cellulaire tumorale , Humains , Souris , Souris nude , Transplantation tumorale , Protéines proto-oncogènes c-bcl-2/métabolisme , Antigènes CD95/métabolismeRÉSUMÉ
OBJECTIVE: To explore the correlation of IGF-1R expression with clinical features of esophageal squamous cell carcinoma (ESCC) and to investigate the effect of silencing IGF-1R by siRNA on the proliferation of esophageal cancer cell line EC9706 cells. METHODS: Immunohistochemistry was used to detect the expresion of IGF-1R in 80 specimens of ESCC and 18 specimens of normal esophageal mucosa. IGF-1R siRNA was transfected into esophageal squamous cell carcinoma EC9706 cells, and the effect of RNAi was assessed by Western blot. The proliferation of EC9706 cells was determined by drawing growth curve, MTT assay and plate colony-forming assay. RESULTS: The total and strong positive rates of IGF-1R expression were 86.3% and 51.3% in ESCC, and 61.1% and 11.1% in normal esophageal epithelium, respectively. The total and strong positive rates of IGF-1R expression in patients with lymph node metastasis were 94.4% and 74.1%, significantly higher than 69.2% and 3.9%, respectively, in those without lymph node metastasis (P<0.01). A significantly higher IGF-1R expression was associated with lower histological grade (P<0.05). The total and strong rates of IGF-1R expression in 39 patients of stages III and IV were 97.4% and 71.8% , significantly higher than the 75.6% and 31.7%, respectively, in 41 cases of stages I and II (P<0.01). IGF-1R RNAi significantly inhibited IGF-1R expression and the growth of EC9706 cells. The clone formation rate of RNAi-IGF-1R transfected cells was 19.1%, significantly lower than that of 52.3% in non-transfected cells and 49.0% in empty vector-transfected EC9706 cells (P<0.05). CONCLUSIONS: The overexpression of IGF-1R is colerated with lymph node metastasis, differentiation and clinical stage. Down-regulation of IGF-1R can inhibit the proliferation of esophageal cancer EC9706 cells in vitro.
Sujet(s)
Carcinome épidermoïde/anatomopathologie , Prolifération cellulaire , Tumeurs de l'oesophage/anatomopathologie , Interférence par ARN , Petit ARN interférent/génétique , Récepteur IGF de type 1/métabolisme , Sujet âgé , Carcinome épidermoïde/génétique , Carcinome épidermoïde/métabolisme , Lignée cellulaire tumorale , Régulation négative , Tumeurs de l'oesophage/génétique , Tumeurs de l'oesophage/métabolisme , Femelle , Régulation de l'expression des gènes tumoraux , Humains , Métastase lymphatique , Mâle , Adulte d'âge moyen , Grading des tumeurs , Stadification tumorale , Récepteur IGF de type 1/génétique , TransfectionRÉSUMÉ
OBJECTIVE: To observe the relationship between expression of retinoic acid receptor-ß(RAR-ß) in esophageal squamous cell carcinoma (ESCC) and chemotherapy response. METHODS: Fifty-two cases advanced ESCC patients treated by DDP and 5-FU, DDP 80 mg/m(2), divided into 5 days; 5-FU 375 mg/m(2), d1-5. Immunohistochemistry was used to examine the expression of RAR-ß in ESCC. Fifty cases normal esophageal tissue were used as controls. RESULTS: RAR-ß immunoreactivity was recognized in both cytoplasm and nucleus, RAR-ß positive rate was lower in ESCC compared with normal tissue (61.5% vs 92%, P < 0.05). The 52 cases ESCC patients were treated 228 chemotherapy cycles, the overall response rate (OR) was 71.2%. The OR in RAR-ß positive patients was 84.4% (27/32), significant higher than RAR-ß negative patients 50.0% (10/20) (P < 0.05). The time-to-progression (TTP) for RAR-ß positive patients was 5.9 months, the median survival period was 12.1 months, 2 years survival rate was 56.7%; whereas TTP for RAR-ß negative patients was 2.1 months, the median survival period was 5.8 months, 2 years survival rate was 32.9%. There was significant difference between the 2 groups (P < 0.05). CONCLUSION: RAR-ß protein expression by immunohistochemistry may be a useful indicator to predict the chemotherapy response and clinical outcome for ESCC, meanwhile it may be an avenue for target therapy.
Sujet(s)
Carcinome épidermoïde/métabolisme , Tumeurs de l'oesophage/métabolisme , Récepteurs à l'acide rétinoïque/métabolisme , Adulte , Sujet âgé , Sujet âgé de 80 ans ou plus , Protocoles de polychimiothérapie antinéoplasique , Carcinome épidermoïde/traitement médicamenteux , Tumeurs de l'oesophage/traitement médicamenteux , Femelle , Humains , Mâle , Adulte d'âge moyenRÉSUMÉ
OBJECTIVE: To investigate the impact of all-trans retinoic acid (ATRA) on chemosensitivity to esophageal squamous cell carcinoma EC9706 cells in vitro and its mechanism. METHODS: EC9706 cells were routinely cultured as the control group. The experimental group was divided into three groups. The ATRA group with ATRA in final concentration of 1 µmol/L; the 5-Fu group with 5-Fu in final concentration of 50 mg/L; the combined treatment group with ATRA in final concentration of 1 µmol/L and 5-Fu 50 mg/L. The cell apoptosis was detected by terminal deoxynucleotidy transferase mediated dUTP nick end labelling (TUNEL). The cell cycle and apoptosis were detected by flow cytometry. RESULTS: The results of TUNEL showed that in the combined treatment group appeared a large number of apoptotic cells, and their nuclei were stained brown, with a positive rate of 89.7%. There was a significant difference in the comparison with the ATRA group (38.3%) and 5-Fu group (40.3%) (P < 0.05). The flow cytometry showed that the ATRA + 5-Fu group had a significantly higher apoptosis rate (76.9% ± 2.7%) than that in the ATRA group (38.2% ± 2.6%) and 5-Fu group (45.2% ± 2.3%) (P < 0.05). The ratio of cells in G(1) phase increased in the ATRA + 5-Fu group (83.4% ± 3.0%), significantly higher than (48.2% ± 2.5%) in the ATRA group and (53.2% ± 2.6%) in the 5-Fu group (P < 0.05). The ratio of cells in S + G(2)/M phase was decreased in the ATRA + 5-Fu group, with a significant difference (P < 0.05) when compared with other groups. There was no significant difference between the ATRA group and 5-Fu group (P > 0.05) in the apoptosis rate and the proportion of cells at different phases. CONCLUSION: ATRA can induce apoptosis of esophageal carcinoma EC9706 cells in vitro. The combination of ATRA and 5-Fu may enhance the chemotherapeutic efficacy.
Sujet(s)
Apoptose/effets des médicaments et des substances chimiques , Carcinome épidermoïde/anatomopathologie , Tumeurs de l'oesophage/anatomopathologie , Fluorouracil/pharmacologie , Trétinoïne/pharmacologie , Antimétabolites antinéoplasiques/pharmacologie , Antinéoplasiques/pharmacologie , Cycle cellulaire/effets des médicaments et des substances chimiques , Lignée cellulaire tumorale , Synergie des médicaments , HumainsRÉSUMÉ
OBJECTIVE: To construct a eukaryotic expression vector of stathmin gene and investigate its effect on esophageal squamous cell carcinoma (ESCC) EC9706 cell line. METHODS: Stathmin cDNA coding sequence was amplified by RT-PCR and cloned into a eukaryotic expression vector pcDNA3.1(+). EC9706 cells were transfected with this recombinant plasmid pcDNA3.1-stathmin by lipofectamine. And the stable transfectants were selected with G418 medium. Stathmin protein expression was detected with Western blot in transfected EC9706 cell lines. Morphologic change of stable transfectants was observed under microscope. The proliferation of transfected cells was measured by cell counting, MTT and in vitro formation assay of flat. Flow cytometry was used to detect the cell cycle. Nude mice were adopted to investigate the in vivo tumorigenic characteristics of the transfected cells. RESULTS: A 450 bp coding sequence of stathmin cDNA was amplified by RT-PCR and then cloned into pcDNA3.1(+) plasmid to harvest the recombinant plasmid pcDNA3.1-stathmin. The recombinant plasmid pcDNA3.1-stathmin and blank vector were transfected respectively into EC9706 cells. The up-regulated expression of stathmin protein was validated by Western blot (P < 0.01). Compared with control, EC9706 cells transfected with pcDNA3.1-stathmin appeared swollen and multi-nuclear with a cell mitotic arrest; doubling generation time of pcDNA3.1-stathmin transfectants was prolonged (25 - 28 h); The in vitro cell proliferation ability and clone formation rate (34.5% ± 6.9%) decreased, cell cleavage was blocked at G(2)/M phase (21.7% ± 3.4%) and the oncogenicity of inoculated cells in nude mice decreased (all P < 0.01). CONCLUSIONS: The up-regulated expression of stathmin protein triggered by the recombinant plasmid pcDNA3.1-stathmin can inhibit the proliferation and oncogenicity of ESCC EC9706 cells. This molecule may be a promising therapeutic target in ESCC patients.
Sujet(s)
Carcinome épidermoïde/métabolisme , Tumeurs de l'oesophage/métabolisme , Stathmine/métabolisme , Animaux , Carcinome épidermoïde/génétique , Lignée cellulaire tumorale , Prolifération cellulaire , Tumeurs de l'oesophage/génétique , Femelle , Vecteurs génétiques , Humains , Souris , Souris nude , Plasmides , Stathmine/génétique , Transfection , Régulation positiveRÉSUMÉ
OBJECTIVE: To explore the expression of stathmin gene in esophageal squamous cell carcinoma (ESCC) and its correlation to oncogenesis of ESCC. METHODS: Three ESCC cell lines, 75 ESCC samples, 25 tumor-adjacent samples and 30 normal esophageal mucosa samples were examined for the expression of stathmin mRNA and protein by in situ hybridization and immunohistochemistry, respectively. The correlations of stathmin expression to the clinicopathological features of the patients were analyzed. RESULTS: Overexpression of stathmin mRNA and protein was found in 3 ESCC cell lines EC9706, Eca109 and EC-1, with the positive expression rates exceeding 80%. The positive rates of stathmin mRNA and protein in ESCC samples were 82.7% and 81.3%, respectively. There were significant differences in the relative contents of stathmin mRNA and protein among normal mucosa tissue, tumor-adjacent tissue and cancer tissue (chi2=19.204 and 25.03, respectively, P<0.01). In addition, a positive correlation was noted between stathmin mRNA and protein expressions in ESCC (r=0.413, P=0.000). The relative contents of stathmin mRNA and protein were significantly correlated to the differentiation degree, lymph node metastasis, invasive depth and TNM stage of ESCC (P<0.05). CONCLUSIONS: The expression of stathmin mRNA and protein is upregulated in ESCC with correlation to the differentiation degree, lymph node metastasis, invasive depth and TNM stage of ESCC, suggesting the possible involvement of stathmin in the oncogenesis of ESCC. Combined detection of stathmin mRNA and protein may prove valuable for early diagnosis and prognosis of ESCC, and stathmin may serve as a potential molecular target for biotherapy of the tumor.