Scinderin promotes glioma cell migration and invasion via remodeling actin cytoskeleton

BACKGROUND Glioma is one of the most common intracranial tumors,characterized by invasive growth and poor prognosis.Actin cytoskeletal rearrangement is an essential event of tumor cell migration.The actin dynamics-related protein scinderin(SCIN)has been reported to be closely related to tumor cell migration and invasion in several cancers.AIM To investigate the role and mechanism of SCIN in glioma.METHODS The expression and clinical significance of SCIN in glioma were analyzed based on public databases.SCIN expression was examined using real-time quantitative polymerase chain reaction and Western blotting.Gene silencing was performed using short hairpin RNA transfection.Cell viability,migration,and invasion were assessed using cell counting kit 8 assay,wound healing,and Matrigel invasion assays,respectively.F-actin cytoskeleton organization was assessed using F-actin staining.RESULTS SCIN expression was significantly elevated in glioma,and high levels of SCIN were associated with advanced tumor grade and wild-type isocitrate dehydrogenase.Furthermore,SCIN-deficient cells exhibited decreased proliferation,migration,and invasion in U87 and U251 cells.Moreover,knockdown of SCIN inhibited the RhoA/focal adhesion kinase(FAK)signaling to promote F-actin depolymerization in U87 and U251 cells.CONCLUSION SCIN modulates the actin cytoskeleton via activating RhoA/FAK signaling,thereby promoting the migration and invasion of glioma cells.This study identified the cancer-promoting effect of SCIN and provided a potential therapeutic target for the treatment of glioma.

Evaluation of TAZ expression and its effect on tumor invasion and metastasis in human glioma

Objective:To evaluate the expression TAZ and its role in tumor invasion and metastetsis in human glioma.Methods:The expression of TAZ protein was measured in 48 samples of surgically resected human glioma and 13 samples of normal brain tissues using immunohistochemistry.TAZ was knocked down by a retrovirus-mediated TAZ shRNA in a glioma cell line.SNB19.Transwell cell migration and invasion assays were used to determine migration and invasion of SNB19 cells.Results:The positive expression tale of TAZ protein in glioma tissnes was significantly higher than than in normal brain tissues(79.2%vs.15.4%.P<0.001).Furthermore.clinical analysis suggested thai the positive expression rate of TAZ protein in poorly differentiated tumor tissues was significantly higher as compared with that in well differentiated tissues(96.0%vs.60.9%,P<0.01).TAZ was significantly knocked down by TAZ shRNA(P<0.001),and TAZ knockdown significantly reduced cell migration and invasion(P<0.01.rspectively)in SNB19cells.Conclusions:TAZ protein overexpression is observed in human glioma and its elevated expression is significantly correlated with poor differentiation.TAZ knockdown prominently reduces cell migration and invasion in SNB19 cells,suggesling that TAZ may play a key role in the initiation and progression of human glioma.

Knockdown of HMGB1 improves apoptosis and suppresses proliferation and invasion of glioma cells

Background： Effective methods for managing patients with solitary pulmonary nodules（SPNs） depend critically on the predictive probability of malignancy.Methods： Between July 2009 and June 2011, data on gender, age, cancer history, tumor familial history, smoking status, tumor location, nodule size, spiculation, calcification, the tumor border, and the final pathological diagnosis were collected retrospectively from 154 surgical patients with an SPN measuring 3-30 mm. Each final diagnosis was compared with the probability calculated by three predicted models—the Mayo, VA, and Peking University（PU） models. The accuracy of each model was assessed using area under the receiver operating characteristics（ROC） and calibration curves.Results： The area under the ROC curve of the PU model [0.800; 95% confidence interval（CI）： 0.708-0.891] was higher than that of the Mayo model（0.753; 95% CI： 0.650-0.857） or VA model（0.728; 95% CI： 0.623-0.833）; however, this finding was not statistically significant. To varying degrees, calibration curves showed that all three models overestimated malignancy.Conclusions： The three predicted models have similar accuracy for prediction of SPN malignancy, although the accuracy is not sufficient. For Chinese patients, the PU model may has greater predictive power.Background： Here, we introduced our short experience on the application of a new CUSA Excel ultrasonic aspiration system, which was provided by Integra Lifesciences corporation, in skull base meningiomas resection.Methods： Ten patients with anterior, middle skull base and sphenoid ridge meningioma were operated using the CUSA Excel ultrasonic aspiration system at the Neurosurgery Department of Shanghai Huashan Hospital from August 2014 to October 2014. There were six male and four female patients, aged from 38 to 61 years old（the mean age was 48.5 years old）. Five cases with tumor located at anterior skull base, three cases with tumor on middle skull base, and two cases with tumor on sphenoid ridge.Results： All the patents received total resection of meningiomas with the help of this new tool, and the critical brain vessels and nerves were preserved during operations. All the patients recovered well after operation.Conclusions： This new CUSA Excel ultrasonic aspiration system has the advantage of preserving vital brain arteries and cranial nerves during skull base meningioma resection, which is very important for skull base tumor operations. This key step would ensure a well prognosis for patients. We hope the neurosurgeons would benefit from this kind of technique.Background： The purposes of this study were to explore the effects of high mobility group protein box 1（HMGB1） gene on the growth, proliferation, apoptosis, invasion, and metastasis of glioma cells, with an attempt to provide potential therapeutic targets for the treatment of glioma. Methods： The expressions of HMGB1 in glioma cells（U251, U-87 MG and LN-18） and one control cell line（SVG p12） were detected by real time PCR and Western blotting, respectively. Then, the effects of HMGB1 on the biological behaviors of glioma cells were detected： the expression of HMGB1 in human glioma cell lines U251 and U-87 MG were suppressed using RNAi technique, then the influences of HMGB1 on the viability, cycle, apoptosis, and invasion abilities of U251 and U-87 MG cells were analyzed using in a Transwell invasion chamber. Also, the effects of HMGB1 on the expressions of cyclin D1, Bax, Bcl-2, and MMP 9 were detected. Results： As shown by real-time PCR and Western blotting, the expression of HMGB1 significantly increased in glioma cells（U251, U-87 MG, and LN-18） in comparison with the control cell line（SVG p12）; the vitality, proliferation and invasive capabilities of U251 and U-87 MG cells in the HMGB1 siR NA-transfected group were significantly lower than those in the blank control group and negative control（NC） siR NA group（P〈0.05） but showed no significant difference between the blank control group and NC siR NA group. The percentage of apoptotic U251 and U-87 MG cells was significantly higher in the HMGB1 siR NA-transfected group than in the blank control group and NC siR NA group（P〈0.05） but was similar between the latter two groups. The HMGB1 siR NA-transfected group had significantly lower expression levels of Cyclin D1, Bcl-2, and MMP-9 protein in U251 and U-87 MG cells and significantly higher expression of Bax protein than in the blank control group and NC siR NA group（P〈0.05）; the expression profiles of cyclin D1, Bax, Bcl-2, and MMP 9 showed no significant change in both blank control group and NC siR NA group. Conclusions： HMGB1 gene may promote the proliferation and migration of glioma cells and suppress its effects of apoptosis. Inhibition of the expression of HMGB1 gene can suppress the proliferation and migration of glioma cells and promote their apoptosis. Our observations provided a new target for intervention and treatment of glioma.

SIGNIFICANCE OF THE EXPRESSION OF GREEN FLUORESCENT PROTEIN ON DETECTION OF GLIOMA INVASION IN VIVO

Objective: To investigate the invasion and metastasis of glioma in vivo by xenotransplanted tumor established by implanting C6 glioma cells transfected with green fluorescent protein (GFP) gene in vitro into the brain of SD rats. Methods: C6 cells were transfected with a plasmid vector (pEGEP-N3) containing the GFP gene. Stable GFP-expressing clones were isolated and performed examination by flow cytometry and electron microscope. GFP-expressing cells were stereotactically injected into the brain parenchyma of SD rats to establish xenotransplanted tumor. Four weeks later rats were killed and continuous brain sections respectively were examined by HE staining, immunohistochemistry method and fluorescence microscopy for detection of tumor cell invasion. Xenotransplanted tumor was primarily cultured to determine the storage of exotic GFP gene in vivo. Results: There were not obvious changes in cell cycle and ultrastructure for the cells transfected with GFP gene. C6 cells transfected with GFP gene maintained stable high-level GFP expression in the central nervous system during their growth in vivo. GFP fluorescence clearly demarcated the primary tumor margin and readily allowed for the detection of distant invasion on the single-cell level, which was evidently superior to HE and immunohistochemistry staining. There was not GFP gene loss of transfected cells in vivo. Conclusions: It is suggested that C6 cells transfected with GFP gene can be visualized by fluorescent microscopy after intracranial implantation. This model is an excellent experimental animal model in research on invasion of glioma.

The effects of antisense PTEN gene transfection on the growth and invasion of glioma cells

Objective:To study the effects of antisense PTEN gene on the growth and invasion of glioma cells. Methods:A pcDNA3. 1/Hygro (-) recombinant plasmid containing antisense PTEN gene fragment was constructed. Glioma cells of primary culture were transfected with antisense PTEN gene vector and stably transfected clones were selected. Then, the different growth and invasion abilities and the different MMP9 mRNA expressions of three kinds of cells were observed, including the transfected cells, untransfected cells and the cells transfected with empty vector. Results :The abilities of growth and invasion of the transfected cells and the expressions of MMP9 mRNA were obviously enhanced. Conclusion: Antisense PTEN gene could have a negative impact on the growth and invasion of primary culture glioma cells.

Effects of RNAi-mediated Gene Silencing of LRIG1 on Proliferation and Invasion of Glioma Cells

The effects of RNAi-mediated gene silencing of LRlG1 on proliferation and invasion of the human glioma cell line U251-MG and the possible mechanisms were explored in this study. The plasmids pGenesil2-LRIG1-shRNA1 and pGenesil2-LRIG1-shRNA2 were transfected into U251-MG glioma cells respectively by using Lipofectamine 2000 and the transfected cells in which the LRIG1 expression was stably suppressed were selected by G418. The cells transfected with negative shRNA served as control. The expression levels of LRIG1 mRNA and protein were measured by qRT-PCR and Western blotting, respectively. The cell cycle was analyzed by flow cytometry. The results showed that LRIG1 mRNA expression was reduced by 70% and 58% and LRIG1 protein expression by 58% and 26% in U251-MG cells transfected with pGenesil2-LRIG1-shRNAl and pGenesil2-LRIG1-shRNA2 relative to the negative shRNA-transfected U251-MG cells. The proliferative capacity of the LRIG1 specific siRNA-transfected cells was stronger than that of control cells. Cell cycle analysis showed that silencing LRIG1 significantly increased the percentage of S phase cells and the proliferation index (P<0.01). Moreover, silencing LRIG1 could promote the invasion of U251-MG cells (P<0.05). These findings suggested that LRIG1-targeting siRNA can exert a dramatically inhibitory effect on RNA transcription and protein expression of LRIG1, and LRIG1 down-regulation could promote the proliferation of U251-MG cells, arrest U251-MG cells in S phase, and enhance the invasion of U251-MG cells.

EXPRESSION OF MATRIX METALLOPROTEINASE-2 AND VASCULAR ENDOTHELIAL GROWTH FACTOR IN HUMAN GLIOMA AND THEIR RELATION TO THE INVASION OF THE TUMOR

Objective To study the expression of vascular endothelial growth factor (VEGF) and matrix metalloproteinase-2 (MMP-2) in different grade human glioma. To investigate their relation to the pathological grade and invasion of the tumor. Methods The expression of MMP-2 and VEGF were determined by immunohistochemical technique in 48 cases of human glioma and 10 specimens of normal brain tissue. Results The expression levels of MMP-2 and VEGF in human glioma were positively related to tumor grades (P<0.01), and their expressions in the glioma of grade Ⅲ and Ⅳ were significantly different from those in the glioma of grade Ⅰ-Ⅱand normal brain tissue (P<0.01). The expression of MMP-2 was positively correlated to that of VEGF (P<0.01). Conclusion MMP-2 and VEGF were highly expression in human glioma and were positively related to the tumor grades. The synergic interaction of MMP-2 and VEGF promoted the angiogenesis and invasion of human glioma.

GDNF promotes malignant glioma cells migration and invasion by upregulating serpine 1 expression

Glial cell line derived neurotrophic factor(GDNF)isa neurotrophic factor,which has obvious nutritional effect on a variety of neurons.In glioma,it promotes the migration and invasion of glioma cells.However,uncovering the underlying mechanism of promoting malignant progression of glioma is still a challenging work.Our sequencing results showed that the expression of Serpine 1 was up-regulated in rat C6 cells after GDNF treatment.In the present study,we found that GDNF promoted Serpine 1 expression at the level of mRNA and protein in C6 cells.We also observed that increasing migration and invasion in C6 cells was stimulated by exogenous GDNF.Serpine 1 knockdown significantly inhibited the migration and invasion of C6 cells.Further investigations revealed that the exogenous GDNF treatment increased the phosphorylation of SMAD2.By contrast,the knockdown of Serpine 1 resulted in the opposite effects.

Research on the effects of PIAS3 expression on the invasion of glioma TJ905 cells

Objective To investigate the function and possible mechanisms of PIAS3 expression on the invasion of TJ905 cells. Methods PIAS3 overexpression vectors were constructed and PIAS3 siRNA were chemically synthesized, which were separately transfected into TJ905 cells for upregulation or

PPM1D Silencing by Lentiviral-mediated RNA Interference Inhibits Proliferation and Invasion of Human Glioma Cells

To construct a lentiviral shRNA vector targeting human protein phosphatase 1D magnesium-dependent（PPM1D） gene and detect its effectiveness of gene silencing in human gliomas,specific siRNA targets with short hairpin frame were designed and synthesized.DNA oligo was cloned into the pFU-GW-iRNA lentiviral expression vector,and then PCR and sequencing analyses were conducted to verify the constructs.After the verified plasmids were transfected into 293T cells,the lentivirus was produced and the titer of virus was determined.Real-time quantitative PCR and Western blot were performed to detect the PPM1D expression level in the infected glioma cells.PCR and Western blot analyses revealed the optimal interfering target,and the virus with a titer of 6×10^8 TU/mL was successfully packaged.The PPM1D expression in human glioma cells was knocked down at both mRNA and protein levels by virus infection.The expression of PPM1D mRNA and protein was decreased by 76.3% and 87.0% respectively as compared with control group.The multiple functions of human glioma cells after PPM1D RNA interference were detected by flow cytometry and cell counting kit-8（CCK-8）.Efficient down-regulation of PPM1D resulted in significantly increased cell apoptosis and reduced cell proliferation and invasion potential in U87-MG cells.We have successfully constructed the lentiviral shRNA expression vector capable of stable PPM1D gene silencing at both mRNA and protein levels in glioma cells.And our data gave evidence that the reduced cell growth observed after PPM1D silencing in glioma cells was at least partly due to increased apoptotic cell death.

Decreased invasion ability of hypotaurine synthesis deficient glioma cells was partially due to hypomethylation of Wnt5a promoter

Glioma is one of the lethal central nervous system tumors.The infiltrative and invasive growth nature makes it difficult to identify the boundary between glioma and the normal tissues,resulting in inevitable recurrence after surgery operation.Gliomas do not metastasize,so to prevent the residual tumor from proliferating or invading is a key challenge.Previous report indicated that hypotaurine could facilitate glioma invasion and suppress demethylases’activities.Using a hypotaurine synthesis deficient U251 cell line,we proved that the cells invasion ability was impaired.Gene expression profile analysis exhibited that knocking down one of the key enzymes of hypotaurine synthesis,2-aminoethanethiol dioxygenase(ADO),significantly affected the extracellular matrix-receptor process.Of that process,Wnt5a expression was severely upregulated by decreased intracellular ADO expression.Cells cultured at the presence of hypotaurine showed a decrease in intracellular Wnt5a protein and mRNA levels.This phenotype was due to hypermethylation of Wnt5a promoter,which was most likely the result of hypotaurine’s inhibiting demethylases activities.Collectively,this study demonstrated that hypotaurine synthesis deficient U251 cells were prone to epigenetic modification and Wnt5a seemed to be a tumor suppressor under that circumstance.This tumor suppression effect is warranted to be reevaluated in real tumor samples and the relevant evidence might contribute to develop new glioma interference strategies.

Tumor Suppressor miR-637 Is Associated with Cellular Migration, Invasion, and Glioma Diagnosis

Objective: Abnormal miRNA expression is observed in several human tumors;moreover, normal cell regulation can be disrupted by tumor-suppressive or oncogenic miRNAs. We aimed to investigate the role of miR-637 in gliomas. Methods: We assessed miR-637 expression in 98 and 16 gliomas and non-tumoral brain tissues, respectively, using in situ hybridization. We calculated receiver operating characteristic curves to determine the specificity and sensitivity of miR-637 biomarkers. Next, the effects of miR-637 on glioma cell migration and invasion were determined by using the transwell assay. Candidate target genes were identified through Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis. Results: There was significant miR-637 downregulation in glioma tissues (P < 0.001). Further, it showed potential as a diagnostic biomarker for gliomas. In addition, miR-637 suppressed glioma cell migration and invasion. Conclusions: Our findings suggest that miR-637 inhibits glioma invasion and migration and could be a potential diagnostic marker for gliomas. Future studies should examine the potential mechanisms underlying miR-637 as a diagnostic marker and therapeutic target for gliomas.

Role of nestin in glioma invasion

AIM: To determine the role for the intermediate filament protein nestin in glioma invasion. METHODS: We examined the expression and function of nestin in gliomas(Grades Ⅱ-Ⅳ as defined by the World Health Organization). We determined nestin expression using Immunohistochemical methods. To elucidate nestin's biological function(s), we reduced m RNA levels by 61% and 87% in two glioblastomaderived neurosphere lines using short hairpin RNAs and determined the effect of reduced nestin expression on glioma cell proliferation and invasion using MTS and matrigel migration assays, respectively. We also utilized quantitative real time polymerase chain reaction assaysto determine the effect of reduced nestin expression on the expression of other markers associated with glioma stem cells and their differentiated progenies. RESULTS: We found a significant correlation between nestin immunoreactivity and astrocytoma tumor grade, with 36% of grade Ⅱ, 75% of grade Ⅲ, and 100% of grade Ⅳtumors expressing significant levels of the protein when assessed using immunohistochemistry. Reduction in nestin expression had no effect on cell growth in culture, but did retard the capacity of one line to migrate in-vitro on matrigel. Interestingly, in the line whose migration was not affected, m RNA levels of a second intermediate filament, synemin(also knowns as desmuslin), were elevated following introduction of sh RNA targeting nestin. As synemin was not induced in the line which required nestin for migration, it is a possibility that synemin may compensate for the loss of nestin in this process. CONCLUSION: Nestin expression is prominent in high-grade astrocytomas. Nestin is not required for cell growth but it may, however, be required for cell motility.

Nectin-like Molecule 1 Inhibits the Migration and Invasion of U251 Glioma Cells by Regulating the Expression of An Extracellular Matrix Protein Osteopontin

Objective To investigate the molecular mechanism of nectin-like molecule 1(NECL1) inhibiting the migration and invasion of U251 glioma cells.Methods We infected U251 glioma cells with adeno-nectin-like molecule 1(Ad-NECL1) or empty adenovirus(Ad).Transwell and wound healing assays were performed to observe the migration of U251 cells incubated with the cell supernatant from Ad-NECL1 or Ad infected U251 cells.DNA microarray was applied to screen the gene expression profile after the restoration of NECL1 in U251 glioma cell lines.The differential expression of osteopontin(OPN),a gene related to migration and invasion,was further analyzed with semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR),Western blot,and immunohistochemistry.Results The restoration of NECL1 inhibited migration of U251 cells significantly(P<0.05).Altogether 195 genes were found differentially expressed by microarray,in which 175 were up-regulated and 20 down-regulated,including 9 extracellular matrix proteins involved in the migration of cells.Both mRNA and protein expressions of OPN,the most markedly reduced extracellular matrix protein,were found decreased in U251 cells after restoration of NECL1.Immunohistochemical assay also detected an increase of OPN in glioma tissues,related with the progressing of malignant grade.Conclusion A link might exist between NECL1 and the extracellular matrix protein OPN in inhibiting the migration and invasion of U251 glioma cells.

Relation of Cystatin C and Cathepsin B Expression to the Pathological Grade and Invasion of Human Gliomas

OBJECTIVE To explore the relation of cystatin C and cathepsin B expression to the pathological grade and invasion of human gliomas. METHODS A immunohistochemical method was used to detect the expression of cystatin C and cathepsin B in 57 glioma samples. RESULTS The expression of cystatin C in high-grade （Grade Ⅲ-Ⅳ ）gliomas was significantly weaker than that in low-grade（Grade Ⅰ-Ⅱ, P=0.0001）. On the other hand, the expression of cathepsin B in high-grade gliomas was significantly stronger than that in low-grade （P=0.0001）. Cystatin C expression correlated inversely with cathepsin B expression in gliomas （P=0.01）. CONCLUSION Cystatin C and cathepsin B expression is related to the pathological grade and invasion of gliomas. Combining detection of cystatin C and cathepsin B expressions might provide significant information for clinical assessment of maglignant phenotypes and invasion of gliomas.

Raf/MEK/ERK Signaling Pathway Is Involved in the Inhibition of Glioma Cell Proliferation and Invasion in the Ketogenic Microenvironment

Objective:A high-fat,low-carbohydrate ketogenic diet has been used to treat malignant glioma,in which the Raf/MEK/ERK signaling pathway is overactivated.However,whether the Raf/MEK/ERK signaling pathway is involved in the therapeutic effect of ketone bodies remains unknown.In this study,we investigated the effects of a major ketone body,3-hydroxybutyric acid(3-HBA),on the proliferation and metastasis of malignant glioblastoma cells and the underlying mechanism.Methods:Two human malignant glioblastoma cell lines(U87 and U251)were treated with different concentrations of 3-HBA with or without the Raf inhibitor PAF C-16 for 24 h.Cell proliferation,cell cycle,cell invasion,and phospholipase D1(PLD1)activity were determined.Protein and gene expression levels of Raf/MEK/ERK signaling pathway members were examined.Results:3-HBA significantly decreased cell proliferation,invasion,and intracellular PLD1 activity in both U87 and U251 glioblastoma cell lines.3-HBA treatment significantly increased the proportion of cells in the G1 phase and decreased the proportion of cells in S phase in U87 cells.In the U251 line,the proportion of treated cells in S phase was increased and proportion of cells in G2 was decreased.3-HBA treatment also significantly decreased the protein expression levels of Raf,MEK,p-MEK,ERK,p-ERK,and PLD1 while increasing p53 expression;an effect that was similar to treatment with the Raf inhibitor.Co-treatment of 3-HBA with the Raf inhibitor further enhanced the effects of the 3-HBA in both cell lines.Conclusion:We confirmed that a ketogenic microenvironment can inhibit glioma cell proliferation and invasion by downregulating the expression of PLD1 through the Raf/MEK/ERK signaling pathway.

miR-200a suppresses glioma cell migration and invasion via directly targeting MAGED4

Melanoma-associated antigen D4(MAGED4)is overexpressed in glioma and has been associated with WHO grade and survival.Here we investigated the exact function of MAGED4 during glioma progression and its regulatory mechanism.Cell migration and invasion were evaluated following up-and down-regulation of MAGED4 expression in glioma cell lines U87MG,U251,A172 and SHG44.Furthermore,we combined bioinformatics tools with biological validation assays to determine the miRNA targeting MAGED4.

LncRNA WEE2-AS1 knockdown inhibits the proliferation,migration and invasion of glioma cells via regulating miR-29b-2-5p/TPM3 axis

Glioma is a general malignant tumor with a dismal prognosis.Long noncoding RNAs(lncRNAs)have been implicated in the initiation and processes of tumors.An investigation of the GEPIA database revealed that long noncoding RNA WEE2 antisense RNA 1(WEE2-AS1)is upregulated in glioma tissues compared to normal brain tissues,and validation with quantitative real-time polymerase chain reaction(qRT–PCR)revealed that WEE2-AS1 expression was consistent with the database prediction.Fluorescence in situ hybridization(FISH)assays revealed that WEE2-AS1 was localized primarily in the cytoplasm.Clone formation experiment and EDU assay were used to detect cell proliferation ability,and Transwell assay was used to detect cell migration and invasion ability,Western-blot assay and immunofluorescence were used to determine TPM3 protein level.Functional experiments revealed that the downregulation of WEE2-AS1 impeded cell proliferation,migration,and invasion in glioma cell lines.Furthermore,downregulation of WEE2-AS1 suppressed tumor growth in vivo.Bioinformatics predictions and integrated experiments indicated that WEE2-AS1 promoted tropomyosin 3(TPM3)expression by sponging miR-29b-2-5p.A dual-luciferase reporter assay was conducted to uncover the binding of WEE2-AS1 and miR-29b-2-5p and that of miR-29b-2-5p and TPM3.Additionally,a series of rescue assays showed that WEE2-AS1 promotes proliferation,migration,and invasion by targeting miR-29b-2-5p to regulate TPM3 expression.Ultimately,the results of this study indicate that WEE2-AS1 plays an oncogenic role in glioma and may promote further investigations of the diagnostic and prognostic value of WEE2-AS1 in glioma.

Role of DJ-1-induced PTEN down-regulation in migration and invasion of human glioma cells

Background and Objective: DJ-1, a suppressor of PTEN, promotes metastasis of different tumors, but its function and mechanisms in glioma metastasis remain unclear. This study aimed to investigate the effect of the DJ-1 protein on the migration and invasion of human glioma cells, and to explore potential mechanisms. Methods: The eukaryotic expression vector pEGFP/DJ-1 and small interfering RNA (siRNA) were constructed and transfected into human glioma SWO-38 cells. The expression of DJ-1 and PTEN in SWO-38 cells were detected by Western blot. Cell migration and invasion were detected by transwell assay. Results: After transfection of pEGFP/DJ-1, the expression of DJ-1 (1.28 ± 0.15 vs. 0.89 ± 0.04, P < 0.05) and focal adhesion kinase (FAK) phosphorylation (0.76 ± 0.12 vs. 0.51 ± 0.04, P < 0.05) were increased, whereas the expression of PTEN (0.74 ± 0.2 vs. 1.04 ± 0.14, P < 0.05) was suppressed. After transfection of DJ-1 siRNA, both DJ-1 (0.33 ± 0.04 vs. 0.88 ± 0.06, P < 0.05) and p-FAK levels (0.33 ± 0.01 vs. 0.44 ± 0.05, P < 0.05) were decreased, but PTEN expression (1.1 ± 0.06 vs. 0.81 ± 0.12, P < 0.05) was increased. Transwell assay data showed that pEGFP/DJ-1 transfection promoted SWO-38 cell migration (57.2 ± 6.50 vs. 40.4 ± 5.0, P < 0.05) and invasion (54.6 ± 4.9 vs. 27 ± 6.7, P < 0.05), whereas DJ-1 siRNA transfection inhibited SWO-38 cells migration (54.4 ± 6.9 vs. 73.4 ± 7.6, P < 0.05) and invasion (44.6 ± 5.8 vs. 69.2 ± 9.2, P < 0.05). Conclusion: Over-expression of DJ-1 promotes SWO-38 cell migration and invasion possibly through the DJ-1 and the PTEN/FAK pathway.

Important role of lymphovascular and perineural invasion in prognosis of colorectal cancer patients with N1c disease

BACKGROUND Lymphovascular invasion(LVI)and perineural invasion(PNI)are associated with decreased survival in colorectal cancer(CRC),but its significance in N1c stage remains to be clearly defined.AIM We retrospectively identified 107 consecutive patients who had CRC with N1c disease radically resected at our hospital.Tumors were reviewed for LVI and PNI by one pathologist blinded to the patients’outcomes.Disease-free survival(DFS),overall survival(OS)and cancer-specific survival(CSS)were determined using the Kaplan-Meier method,with LVI and PNI prognosis differences determined by multivariate analysis using the Cox multiple hazards model.Results were compared using log-rank test.The receiver operating characteristic(ROC)curve was used to evaluate the prognostic predictive ability.RESULTS The median follow-up time was 63.17(45.33-81.37)months for DFS,with 33.64%(36/107)of patients experiencing recurrence;21.5%of tumors were found to be LVI positive and 44.9%PNI positive.The 5-year DFS rate was greater for patients with LVI-negative tumors compared with LVI-positive tumors(74.0%vs 35.6%),and PNI was similar(82.5%vs 45.1%).On multivariate analysis,LVI[hazard ratio(HR)=3.368,95%confidence interval(CI):1.628-6.966,P=0.001]and PNI(HR=3.055,95%CI:1.478-6.313,P=0.002)were independent prognostic factors for DFS.All patients could be divided into three groups of patients with different prognosis according to LVI and PNI.The 5-year ROC curve for LVI,PNI and their combination prediction of DFS was 0.646,0.709 and 0.759,respectively.Similar results were seen for OS and CSS.CONCLUSION LVI and PNI could serve as independent prognostic factors of outcomes in N1c CRC patients.Patients with LVI or PNI should be given more attention during treatment.