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At least 307 records · Page 17

MiR-615 inhibits cell proliferation, migration and invasion by targeting EGFR in human glioblastoma

Highlights: • MiR-615 is decreased in GBM. • MiR-615 is a tumor suppressor. • MiR-615 targets EGFR 3′-UTR. MiR-615 and epidermal growth factor receptor (EGFR) are associated with a number of disease processes and pathogenesis. However, little is known about the mechanisms of miR-615 and EGFR in human glioblastoma multiforme (GBM). Here, we found that down-regulation of miR-615 expression occurred in GBM tissues and cells, and was inversely correlated with overall survival, relapse-free survival, WHO grade as well as EGFR expression. We further determined that miR-615 functions as a tumor suppressor by inhibiting GBM cell proliferation, cell cycle, migration and invasion, and promoting cell apoptosis. In-vivo assay validated the inhibition effect of miR-615 on tumor growth and EGFR expression. Luciferase reporter assays demonstrated that miR-615 targeted the 3′-untranslated region (3′-UTR) of EGFR. Besides, over-expression of EGFR reversed the inhibition effects of miR-615, while silencing of EGFR aggravated these inhibition effects. In conclusions, we identified that miR-615 plays a tumor suppressor role in GBM cell proliferation, migration and invasion by targeting EGFR expression, and miR-615 may act as a novel biomarker for early diagnosis or therapeutic targets of GBM.

60 APPLIED LIFE SCIENCES↗

Knockout of NCOA5 impairs proliferation and migration of hepatocellular carcinoma cells by suppressing epithelial-to-mesenchymal transition

Highlights: • Generation of NCOA5-knockout HCC cell lines using CRISPR/Cas9. • Knockout of NCOA5 inhibits the proliferation and migration of HCC cells. • Knockout of NCOA5 depresses the tumor microsphere formation of HCC cells. • Knockout of NCOA5 suppresses EMT. Nuclear receptor coactivator 5 (NCOA5) plays important roles in the development of a variety of malignancies. However, the underlying mechanisms remain obscure. In this study, we successfully generated the NCOA5 knockout hepatocellular carcinoma (HCC) cells by CRISPR/Cas9 - mediated genome editing and found that knockout of NCOA5 inhibited the proliferation and tumor microsphere formation of HCC cells significantly. Moreover, the migration ability of NCOA5 knockout HCC cells declined. Mechanistic analyses indicated that knockout of NCOA5 can suppress the epithelial - mesenchymal transition (EMT) in HCC cells. In conclusion, our findings provide a mechanistic insight into the role of NCOA5 in HCC progression.

60 APPLIED LIFE SCIENCES↗

Loss of nucleolar localization of NAT10 promotes cell migration and invasion in hepatocellular carcinoma

Highlights: • Residues 68–75 and 989–1018 are complete NuLSs of NAT10. • NuLSs-deleted NAT10 translocates from nucleolus to cytoplasm and membrane. • NuLSs-deleted NAT10 facilitates α-tubulin acetylation, stabilizes microtubule and promotes HCC cell migration. • Cytoplasmic and membranous NAT10 is correlated with poor survival of HCC patients. NAT10, a nucleolar acetyltransferase, participates in a variety of cellular processes including ribosome biogenesis and DNA damage response. Immunohistochemistry staining showed that cytoplasmic and membranous NAT10 is related to the clinical pathologic characteristics in human cancer tissues. However, the mechanism about how NAT10 translocates from the nucleolus to cytoplasm and membrane is unclear. Here, we obtain a NAT10 deletion mutant localizing in cytoplasm and membrane. Bioinformatics analysis showed that residues 68–75 and 989–1018 are two potential nuclear localization signals (NLS) of NAT10. GFP-NAT10 deletion mutant (Δ989-1018) predominantly translocates into cytoplasm with faint signal retained in the nucleolus, while GFP-NAT10(Δ68-75) still remains in the nucleolus and nucleoplasm, indicating residues 989–1018 is the main nucleolar localization signal (NuLS). GFP-NAT10-D3, with both fragments (residues 68–75 and 989–1018) deleted, completely excludes from the nucleolus and translocates to cytoplasm and membrane. Therefore, complete NuLSs of NAT10 should include residues 68–75 and 989–1018. The cytoplasmic and membranous NAT10 mutant (Flag-NAT10-D3) colocalizes with α-tubulin in cytoplasm and with integrin on cell membrane. Importantly, Flag-NAT10-D3 promotes α-tubulin acetylation and stabilizes microtubules. Consequently, Flag-NAT10-D3 promotes migration and invasion in hepatocellular carcinoma (HCC) cells. Statistical analysis of immunohistochemistry staining of NAT10 in HCC tissues demonstrates that the cytoplasmic NAT10 is correlated with poorer prognosis compared with nuclear NAT10, while the membranous NAT10 predicts the poorest clinical outcome of the patients. We thus provide the evidence for the function of cytoplasmic and membranous NAT10 in the metastasis and prognosis of HCC patients.

60 APPLIED LIFE SCIENCES↗

HOXC10 promotes gastric cancer cell invasion and migration via regulation of the NF-κB pathway

Homeobox gene C10 (HOXC10), known to regulate cell differentiation and proliferation, is upregulated in gastric cancer (GC). The mechanism underlying HOXC10 involvement in GC metastasis is unclear. Herein, we found that HOXC10 is overexpressed in GC relative to normal controls. In the 73 GC patients tissue microarrays (TMA) tested, HOXC10 expression was closely related with tumor-node-metastasis (TNM) stage, lymph node metastasis, and distant metastasis. HOXC10 overexpression tended to associate with low 5-year cumulative survival. Cox regression analysis identified HOXC10 as an independent prognostic factor for poor patient survival. HOXC10 promoted GC cell invasion and migration, regulated the expression of activated ATM and markers of NF-κB signaling. HOXC10 may promote invasion and migration of GC by regulating ATM/NF-κB signaling pathway. HOXC10 should be explored as a clinical marker of GC prognosis.

60 APPLIED LIFE SCIENCES↗

SIRT1 promotes proliferation, migration, and invasion of breast cancer cell line MCF-7 by upregulating DNA polymerase delta1 (POLD1)

Highlights: • SIRT1 is increased in clinical breast cancer tissues. • SIRT1 is upregulated in breast cancer cell line MCF-7. • SIRT1 and p125 are positively correlated in clinical breast cancer tissues. • SIRT1 can upregulate the expression level of POLD1/p125 in MCF-7 cells. Sirtuin 1 (SIRT1), class III histone deacetylase, plays an important character in cell proliferation, cell cycle, apoptosis, energy metabolism and DNA repair. In recent years, researchers have attached increasing attention on the role of SIRT1 in tumorigenesis, development and drug resistance. The effect of SIRT1 on breast cancer is still controversial and its exact role remains to be elucidated. In the present study, we investigated the significant role of SIRT1 in breast cancer by exploring the effect of SIRT1 on DNA polymerase delta1 (POLD1), the gene coding for DNA polymerase δ catalytic subunit p125. Immunohistochemistry showed that the protein expression level of SIRT1 was higher in breast cancer tissues relative to adjacent normal tissues. Knockdown of SIRT1 by shRNA decreased the proliferation, migration, and invasion of human breast cancer cell line MCF-7, while the overexpression of SIRT1 promoted the proliferation, migration, and invasion of MCF-7 cells. Clinically, the immunohistochemistry results revealed that the expression of SIRT1 was positively correlated with p125. Further analysis demonstrated that silencing of SIRT1 increased the expression of p53, while the expression level of POLD1/p125 decreased, and the result by overexpressing SIRT1 was opposite. Collectively, these data suggest that SIRT1 is an oncogenic factor in breast cancer cells and can be involved in the progression of breast cancer by inhibiting p53 and activating POLD1. Our finding provides new insights into the mechanisms of breast cancer.

60 APPLIED LIFE SCIENCES↗

The peptide mimicking small extracellular loop domain of CD82 inhibits tumor cell migration, adhesion and induces apoptosis by inhibiting integrin mediated signaling

Highlights: • The peptide mimicking EC1 amino acid sequence (EC1-mP) has the functional role of CD82 intact molecule. • EC1-mP inhibits cancer cell migration, adhesion and induces cancer cells apoptosis. • EC1-mP surpresses the expression of integrin α5 and β1, inhibits integrin mediated signaling. • This novel finding will improve our understanding of the mechanism by which CD82 inhibits metastasis, . • EC1-mP may be a promising candidate for developing anti-metastasis drugs. Within the extracellular domains of metastasis suppressor CD82, the large extracellular loop (EC2) has received much of the attention and its structure and function have been studied in detail. However, little attention has been given to the small extracellular loop (EC1 domain). To investigate the function role of EC1 in metastasis suppression of CD82, the peptide mimicking EC1 amino acid sequence (EC1-mP) was synthesized and its effect on cancer cells behavior was examined. Here, we reported that EC1-mP strongly inhibited cancer cell migration in vitro, attnuated the ability of cancer cells adhesion to fibronectin, and induced the apoptosis. Furthermore, the EC1-mP was showed to supprese the expressions of integrins α5 and β1, as well as decreased the phosphorylation of FAK and expression of ILK in SW620 cells. Taken together, these results demonstrate that this small peptide has the functional role of CD82 intact molecule. This novel finding will improve our understanding of the mechanism by which CD82 inhibits metastasis, and suggested that EC1 mimic peptide may be a promising candidate for developing anti-metastasis drugs.

60 APPLIED LIFE SCIENCES↗

ZKSCAN3 promotes breast cancer cell proliferation, migration and invasion

Highlights: • ZKSCAN3 is upregulated in breast cancer tissue and cell lines. • Knocking down ZKSCAN3 suppresses breast cancer cell proliferation, enhances cell apoptosis and blocks cell cycle. • Knocking down ZKSCAN3 significantly inhibits breast cancer cell migration and invasion. • Knocking down ZKSCAN3 significantly inhibits breast tumor growth in vivo. • Knocking down ZKSCAN3 reduces activity of the Akt/mTOR pathway. ZKSCAN3, a zinc-finger transcription factor, which has been shown to be upregulated in several human cancer. However, the expression level, function and mechanism of ZKSCAN3 in breast cancer remains unknown. In the current study, immunohistochemistry, western blot and quantitative real time polymerase chain reaction (qRT-PCR) results showed that ZKSCAN3 was overexpressed in breast cancer tissue compared with normal breast tissue. Through analyzing the clinicopathological characteristics, we demonstrated that positive ZKSCAN3 expression predicted poor prognosis of patients with breast cancer. The expression level of ZKSCAN3 protein/mRNA in breast cancer cells (MCF-7 and MDA-MB-231) was higher than its expression in normal breast cells (HBL-100). Knocking down ZKSCAN3 via its short hairpin RNA (shRNA) in MCF-7 and MDA-MB-231 inhibited cell viability, migration and invasion. Western blot analysis showed that ZKSCAN3 silencing lead to significant decreases in the expression of Cyclin D1, B-cell lymphoma-2 (Bcl-2), and matrix metalloproteinase (MMP)-2/MMP-9, as well as increases in the expression of Bcl2 Associated X Protein (Bax) in breast cancer cells. Additionally, ZKSCAN3-shRNA expression markedly suppressed tumor growth in breast cancer xenograft mice. Finally, we demonstrated that silencing of ZKSCAN3 was able to inhibit Akt/mTOR signaling pathway by blocking p-Akt and p-mTOR protein expression in breast cancer cells. These results demonstrate that ZKSCAN3 plays a significant role in the progression of breast cancer. Therefore, ZKSCAN3 is a potential therapeutic target for breast cancer.

60 APPLIED LIFE SCIENCES↗

Interleukin-22 promotes triple negative breast cancer cells migration and paclitaxel resistance through JAK-STAT3/MAPKs/AKT signaling pathways

Highlights: • The “only IL-22” producing (Th22) cells were gradually increased in normal, paratumor and tumor tissues of TNBC patients. • IL-22 promotes TNBC cells (MDA-MB 231 and MDA-MB 468) migration and induces paclitaxel resistance. • IL-22 induces TNBC cells migration and paclitaxel resistance through JAK-STAT3/MAKPs/AKT signaling pathways. Owing to the absence of any targeted therapies, triple negative breast cancer (TNBC), which accounts for approximately 15% of all breast cancers, typically have a poorer outcome. In an attempt to find out the TNBCs' microenvironment, we ran into the endogenous expression of a newly discovered cytokine known as Interleukin (IL)-22.

60 APPLIED LIFE SCIENCES↗

c-Fos is necessary for HGF-mediated gene regulation and cell migration in Schwann cells

Highlights: • HGF up-regulates c-Fos expression in distal SCs in injured sciatic nerves. • HGF increase c-Fos expression in primary Schwann cells. • HGF induce c-Fos expression in SCs by ERK/CREB pathway. • c-Fos protein has role in various HGF-mediated responses in SCs. We previously reported that the expression of hepatocyte growth factor (HGF) was highly induced after peripheral nerve damage, and that c-Fos is one of many cellular genes whose expressions are affected by the increased level of HGF[1]. c-Fos is an important component of AP-1 heterodimer, but its role has not been clearly understood in the context of HGF and Schwann cells (SCs). In this study, we investigated the relationship between HGF and c-Fos. First, it was confirmed that the c-Fos was increased in SCs after nerve injury, while this effect abrogated by PHA-665752, an inhibitor of c-met receptor. When primary SCs were treated with recombinant HGF protein, c-Fos expression was regulated in a typical quick, transient fashion at both RNA and proteins levels. HGF-mediated induction of c-Fos expression was highly suppressed by specific inhibitors of ERK and CREB, respectively. The knock down of c-Fos expression by siRNA almost completely blocked various HGF-mediated effects in SCs, such as induction of gene expression of GDNF, LIF, and c-Myc, and migration of SCs, indicating that c-Fos might play a key role in HGF effects. Taken together, our results suggested that c-Fos plays a key role(s) in HGF-mediated effects on neurotrophic genes and cell migration.

60 APPLIED LIFE SCIENCES↗

MicroRNA-613 promotes colon cancer cell proliferation, invasion and migration by targeting ATOH1

The aim of the present study is to investigate the expression and function of miR-613 in colon cancer (CC) and illuminate the molecular mechanisms underlying miR-613-regulated CC progression. Our data demonstrated that miR-613 was upregulated in CC tissue samples (P = 0.009) and human CC cell lines (HCT-116 and Lovo; P = 0.001 and P = 0.003, respectively), which also promoted the proliferation, invasion and migration of CC cells (P < 0.05). The dual-luciferase reporter assay confirmed that Atonal homolog1 (ATOH1) was the target mRNA of miR-613. Rescue experiments showed that ATOH1 overexpression vector significantly reversed the stimulative effects of miR-613 mimic on the progression of HCT-116 and Lovo cells (P < 0.001). Positive ATOH1 expression in CC tissues was significantly associated with lower grade (χ2 = 3.592, P = 0.043), lower TNM stage (χ{sup 2} = 3.537, P = 0.048) and better overall survival (P=0.041). Jun N-terminal kinase 1 (JNK1) pathway and Mucin 2 (MUC2) were the potential downstream proteins of miR-613/ATOH1. miR-613 is an oncogene in CC and promotes the proliferation, invasion and migration of CC cells by targeting ATOH1 likely via activating JNK1 pathway and upregulating MUC2.

60 APPLIED LIFE SCIENCES↗

Upregulation of 24(R/S),25-epoxycholesterol and 27-hydroxycholesterol suppresses the proliferation and migration of gastric cancer cells

Highlights: • Detection of oxysterols in human gastric tumour tissues and gastric juice. • 24(R/S),25-EC and 27HC were increased in human gastric tumour tissues. • Proliferation of gastric cancer cells were suppressed by 24(R/S),25-EC/27HC. • Migration of gastric cancer cells were suppressed by 24(R/S),25-EC/27HC. • Roles of LXRs in the regulation of proliferation/migration by 24(R/S),25-EC/27HC. Gastric cancer (GC) is one of the most common cancers and is the second-leading cause of cancer-associated morbidity worldwide. Oxysterols are oxidized derivatives of cholesterol that may be important in many biological processes, but the levels and roles of oxysterols in gastric tumours remain to be elucidated. The levels of cholesterol, oxysterols and sulfated oxysterols in human gastric tumour tissues, adjacent normal mucosal tissues, cancerous gastric juice and gastric juice obtained from healthy subjects were detected by LC-MS. It was found that the levels of 24(R/S),25-EC and 27HC in human gastric tumour tissues and cancerous gastric juice were significantly increased compared with those of adjacent normal mucosal tissues and gastric juice from healthy subjects. Compared with normal gastric mucosal tissue, the levels of sulfated 25-hydroxycholesterol (25HC3S) and the ratio of 25HC3S/25HC were decreased in human gastric tumour tissues, which might be related to the dramatically decreased SULT2A1 expression in gastric tumour tissue. Both 24(R/S),25-EC and 27HC suppressed gastric cancer proliferation, which was not altered by LXRα-siRNA treatment. The suppression of cell proliferation induced by 27HC was attenuated by LXRβ-siRNA, but the suppression of cell proliferation induced by 24(R/S),25-EC was intensified by LXRβ-siRNA. Both 24(R/S),25-EC and 27HC dramatically inhibited HGC-27 cell migration, which was attenuated by the co-transfection of cells with LXRα-siRNA and LXRβ-siRNA, but not LXRα-siRNA or LXRβ-siRNA alone. In conclusion, the accumulated 24(R/S),25-EC and 27HC in human gastric tumour tissues might play important roles in gastric cancer development.

60 APPLIED LIFE SCIENCES↗

Hsa{sub c}ircRNA{sub 1}03809 regulated the cell proliferation and migration in colorectal cancer via miR-532–3p / FOXO4 axis

Highlights: • The expression of hsa{sub c}ircRNA{sub 1}03809 is down-regulated in the CRC tissues and cells. • Knockdown of hsa{sub c}ircRNA{sub 1}03809 promotes cell proliferation in CRC. • Knockdown of hsa{sub c}ircRNA{sub 1}03809 promotes cell migration in CRC. • Hsa{sub c}ircRNA{sub 1}03809 acts as a suppressor via regulation of miR-532–3P/FOXO4 axis in CRC. Circular RNAs(circRNAs) are a class of non-coding RNAs that are widely expressed in a variety of cell species. The role they play in cancers is poorly understood, especially in colorectal cancer (CRC). Hsa{sub c}ircRNA{sub 1}03809 (hsa{sub c}irc{sub 0}072088, circZFR)has been demonstrated to be lowly expressed in colorectal cancer tissues and is associated with stage and lymph node metastasis of cancer tissues. Real-time quantitative PCR (qRT-PCR) was used to verify the relationship of hsa{sub c}ircRNA{sub 1}03809 between colorectal cancer and paired adjacent tissue in clinical tissue samples. Then, the proliferative capacity, migration ability, cell cycle, and apoptosis were measured using wound-healing assay, CCK8, transwell assay, flow cytometry, and the like, when hsa{sub c}ircRNA{sub 1}03809 expression in SW620 and COCA-2. The qRT-PCR, western bolt and other experiments verify that the expression of hsa{sub c}ircRNA{sub 1}03809 can regulate the expression of miR-532–3P and FOXO4. Hsa{sub c}ircRNA{sub 1}03809 was found to be significantly down regulated in CRC tissues and cell lines and compared with paired adjacent non-tumorous tissues and normal FHC cells. Hsa{sub c}ircRNA{sub 1}03809 participates in the regulation of biological functions through the miR-532–3P/FOXO4 axis in the CRC. Hsa{sub c}ircRNA{sub 1}03809 may be a potential novel gene target for the diagnosis and treatment of CRC.

60 APPLIED LIFE SCIENCES↗

Defective rapid cell shape and transendothelial migration by calpain-1 null neutrophils

Highlights: • The role of calpain-1 in neutrophil shape change was investigated. • Neutrophils were obtained from knock-out mice and assessed ex vivo. • These cells failed to migrate through endothelial monolayers. • Spontaneous cell spreading was grossly abberant. • Calpain-1 null cells failed to spread in response to IP{sub 3} -induced Ca{sup 2+} signals. It has been proposed that Ca{sup 2+} activation of calpain-1 is an important trigger for rapid cell spreading by neutrophils. In this paper, we have investigated this by assessing the ex vivo functioning of neutrophils from calpain-1 null mice, Calpain-1 null neutrophils failed to migrate through TNF-activated endothelial monolayers. The failure to transmigrate through endothelial monolayers was therefore unlikely to be due to a failure of chemotaxis as chemotaxis by adherent calpain-1 null neutrophils towards fMLP was unpaired. In contrast, the capacity of calpian-1 neutrophils to spontaneously spread was limited to smaller diameters than for wild type cells. Photolytic uncaging of IP{sub 3} with Individual wild type neutrophils resulted in a large Ca{sup 2+} signal and rapid cell spreading. In contrast, calpain-1 neutrophils failed to spread in response to the IP{sub 3}-induced Ca{sup 2+} signal. This work has therefore demonstrated that the presence of calpain-1 was required for effective rapid cell spreading by neutrophils.

60 APPLIED LIFE SCIENCES↗

Migration energy barriers and diffusion anisotropy of point defects on tungsten surfaces

We report tungsten is one of the most promising candidates for plasma-facing materials in future fusion devices, owing to its high performance under extreme irradiation conditions. However, irradiation-induced surface morphology varies significantly, depending on the irradiation type, fluence, and flux. Therefore, it is critical to examine the dynamics of point defects on tungsten (W) surfaces to understand how irradiation affects surface morphology. In this study, we employ the Self-Evolving Atomistic Kinetic Monte Carlo (SEAKMC) method to search for potential migration paths of point defects on W (1 0 0), (1 1 0) and (1 1 1) surfaces. The first-principles calculations are then used to accurately determine the migration energy barriers. The obtained paths and barriers are incorporated into a kinetic Monte Carlo (KMC) model to determine trajectories and diffusivities, which are described by diffusion tensors to demonstrate their anisotropic features. Multiple diffusion mechanisms have been identified on different surfaces, with various anisotropy factors. Particularly, point defects on the W (1 1 0) surface have the highest diffusivities, with anisotropy factors independent of temperatures. In comparison, the anisotropy of the W (1 0 0) surface decreases as temperature increases, while the W (1 1 1) surface is isotropic for point defects. This study provides insights into defect transport properties on different surfaces, which are essential for understanding the early stages of irradiation-induced microstructural evolutions and surface morphology of tungsten.

36 MATERIALS SCIENCE↗

Particle Migration in Large Cross-Section Ceramic On-Demand Extrusion Components

Ceramic On-Demand Extrusion (CODE) is a direct ink writing process which allows for the creation of near theoretically dense ceramic components with large cross-sections due to oil-assisted drying. Here, Yttria-stabilized zirconia (YSZ) colloidal pastes were used in CODE to produce dense (multi-road infill and ≳ 98% relative density), large continuous volume (> 1 cm 3 ), and high fidelity (nozzle diameters ≲ 1 mm) structural ceramic components with nanoparticle feedstocks (~d 50 ≲ 1 µm). However, many of these printed components underwent significant particle migration after forming. The reason for this particle migration defect was investigated using the coffee-ring effect for dilute solutions and rheological methods for dense suspensions. Modifications to the colloidal paste, such as changes in solids loading, pH, or surfactant concentration were explored as to their effectiveness to mitigate the defect. Ultimately, paste formulation and printing trade-offs are discussed with respect to the post-printing defect and as to general direct-write patterning.

36 MATERIALS SCIENCE↗

Interstitial migration behavior and defect evolution in ion irradiated pure nickel and Ni-xFe binary alloys

Transition from long-range one-dimensional to short-range three-dimensional migration modes of interstitial defect clusters greatly reduces the damage accumulation in single-phase concentrated solid solution alloys under ion irradiation. A synergetic investigation with experimental, computational and modeling approaches revealed that both the resistance to void swelling and the delay in dislocation evolution in Ni-Fe alloys increased with iron concentration. This was attributed to the gradually increased sluggishness of defect migration, which enhances interstitial and vacancy recombination. Transition from long-range one-dimensional defect motion in pure nickel to short-range three-dimensional motion in concentrated Ni-Fe alloys is continuum, not abrupt, and within an iron concentration range up to 20%. Lastly, the gradual transition process can be quantitatively characterized by the mean free path of the interstitial defect clusters.

36 MATERIALS SCIENCE↗

Overexpression of miR-133 decrease primary endothelial cells proliferation and migration via FGFR1 targeting

Highlights: • FGFR1 signaling can bring about angiogenesis in cancer. • Through downregulating FGFR1, miR-133 caused reductions in bFGF activity. • Reduced bFGF activity result in diminished proliferation and migratory ability of ECs. • Overexpression of miR-133 could reduce the rate of cell division. • MiR-133 could disturb capillary network formation of transfected ECs as well. Angiogenesis is one of the essential hallmarks of cancer that is controlled by the balance between positive and negative regulators. FGFR1 signaling is crucial for the execution of bFGF-induced proliferation, migration, and tube formation of endothelial cells (ECs) and onset of angiogenesis on tumors. The purpose of this study is to identify whether or not miR-133 regulates FGFR1 expression and accordingly hypothesize if it plays a crucial role in modulating bFGF/FGFR1 activity in ECs and blocking tumor angiogenesis through targeting FGFR1. The influences of miR-133 overexpression on bFGF stimulated endothelial cells were assessed by cell growth curve, MTT assaying, tube formation, and migration assays. Forced expression of miR-133 caused significant reductions in bFGF-induced proliferation and migratory ability of ECs. MiR-133 Expression was negatively correlated with both mRNA and protein levels of FGFR1 in the transfected ECs isolated from peripheral blood. Moreover, overexpression of miR-133 drastically reduced the rate of cell division and disturbed capillary network formation of transfected ECs. These findings suggest that miR-133 plays an important function in bFGF-induced angiogenesis processes in ECs and provides a rationale for new therapeutic approaches to suppress tumor angiogenesis and cancer.

60 APPLIED LIFE SCIENCES↗

ADAM17 promotes cell migration and invasion through the integrin β1 pathway in hepatocellular carcinoma

ADAM17 is believed to promote tumor development by facilitating both cell proliferation and migration. In this study, we investigated the involvement of ADAM17 and the activation of the integrin pathway in the regulation of the malignant properties of hepatocellular carcinoma cells and tissues. ADAM17 was positively correlated with active integrin β1, which was determined using a human tissue microarray and an N-nitrosodiethylamine-induced HCC mouse model. We found elevated ADAM17 and active integrin β1 levels in HCC tissues compared with adjacent liver tissues, and the active integrin β1 levels were associated with tumor size and TNM grade. High ADAM17 and active integrin β1 levels in tumor tissues were significantly associated with poor survival of HCC patients. RNAi-mediated ADAM17 knockdown and integrin β1 blockade significantly attenuated the migration and invasion of HCC cells, and overexpression of ADAM17 showed the reverse effects. ADAM17 interference attenuated the intrahepatic growth and metastasis of HCC cells in an orthotopic xenograft model. ADAM17-knockdown cells showed diminished levels of active integrin β1, p-FAK, p-AKT, MMP-2 and MMP-9. ADAM17 knockdown significantly attenuated the translocation of the Notch1 intracellular domain into the nucleus, whereas overexpression of the Notch1 intracellular domain rescued the translocation and enhanced the activation of integrin β1. Our data provide evidence for ADAM17 as an important determinant of malignant properties via regulation of integrin β1 activation and Notch1 signaling. Inhibition of ADAM17 may provide viable therapeutic potential for preventing HCC metastasis.

60 APPLIED LIFE SCIENCES↗