Effects of silencing AEG-1 gene by shRNA on vasculogenic mimicry of a glioma xenograft model / 西安交通大学学报(医学版)

【Objective】 To investigate the effects of silencing AEG-1 gene by shRNA on vasculogenic mimicry (VM) of a glioma xenograft model. 【Methods】 U87 glioma cells were infected with AEG-1 shRNA lentivirals. Real-time PCR and Western blotting were used to detect the mRNA and protein expressions of AEG-1 in U87 cells after infected by the AEG-1 shRNA lentivirals. A glioma xenograft model was generated and CD 34/PAS double-staining was performed to detect the VM channels in vivo. The immunohistochemical assay was performed to evaluate the expressions of MMP-2, MMP-9, VE-cadherin, and VEGF in glioma xenograft models. 【Results】 AEG-1 shRNA lentivirals could significantly inhibit the AEG-1 expression in glioma cells (P<0.01). Meanwhile, they also decreased the number of VM in the glioma xenograft model (P<0.01). Furthermore, the expressions of MMP-2, MMP-9, VE-cadherin, and VEGF in glioma significantly decreased in vivo (P<0.01). 【Conclusion】 These results suggest that silencing AEG-1 gene by shRNA can significantly inhibit VM of glioma in vivo, the mechanism of which may partly be through regulating MMP-2, MMP-9, VEGF, and VE-cadherin expressions.

Effects of down-regulation of astrocyte elevated gene-1 expression by siRNA on glioma vasculogenic mimicy / 西安交通大学学报(医学版)

Objective: To investigate the effects of down-regulation of astrocyte elevated gene-1(AEG-1) expression by siRNA on glioma vasculogenic mimicy in vitro. Methods: The expression of AEG-1 in U87 glioma cells was down-regulated by siRNA. An in vitro model of glioma vasculogenic mimicy was established to evaluate the effect of down-regulation of AEG-1 expression on glioma vasculogenic mimicy. Real-time PCR and Western blot were used to detect the mRNA and protein expressions of VEGF and MMP2 in U87 cells after treated with AEG-1 siRNA, respectively. Immunofluorescence was used to detect the expressions of VEGF and MMP2 in glioma vasculogenic mimicy model in vitro. Results: Down-regulation of AEG-1 expression by siRNA could significantly inhibit the glioma vasculogenic mimicy in vitro(P<0.01). Meanwhile, It could also inhibit the expressions of VEGF and MMP2 in glioma cells and glioma vasculogenic mimicy in vitro (P<0.01). Conclusion: These results suggest that down-regulation of AEG-1 expression by siRNA can significantly inhibit the glioma vasculogenic mimicy in vitro, and these effects may be realized partly through regulation of VEGF and MMP2 expression.

Mutational screening of the negative regulatory region of astrocyte elevated gene-1 promoter in hepatocellular carcinoma patients / Triagem mutacional da região reguladora negativa do promotor do gene-1 elevado em astrocito em pacientes com carcinoma hepatocelular

Astrocyte elevated gene-1 (AEG-1) is a positive regulator of tumorigenesis in human cancer cells. Human AEG-1 gene is located in chromosome 8q22 having 12 exons/11 introns. Chromosome 8q22 is known to be a hot spot for genomic alterations in several cancerous cells involving HCC. The aim of the study was assess association between the negative regulatory region of AEG-1 promoter mutations and genetic susceptibility to hepatocellular carcinoma. The negative regulatory region of the human AEG-1 promoter was evaluated in a total of 50 Iranian hepatocellular carcinomas (HCC) patients. For investigating AEG-1 promoter polymorphisms the PCR-sequencing method was used. In this study was found two new mutation C>T (-633) and G>C (-660) in the patient group. But it was not revealed the statistically significant association between any mutations in this region of the AEG-1 promoter with HCC susceptibility. According to presented data, we can say that the negative regulatory region of the AEG-1 promoter mutations did not exihibit significant relevance with hepatocellular carcinoma. We recommend further studies on the efficacy of the AEG-1 promoter in therapeutic targeting of the HCC.

Resumo: O gene AEG-1 é um regulador positivo da tumorigênese em células cancerígenas humanas. O gene humano AEG-1 está localizado no cromossomo 8q22 com 12 exons/11 introns. O cromossomo 8q22 é conhecido por ser um hotspot para alterações genômicas em várias células cancerígenas que envolvem o CHC. O objetivo do estudo foi avaliar a associação entre a região reguladora negativa das mutações do promotor AEG-1 e a suscetibilidade genética ao carcinoma hepatocelular. A região reguladora negativa do promotor humano AEG-1 foi avaliada em um total de 50 pacientes iranianos com carcinomas hepatocelulares (CHC). Para investigar os polimorfismos do promotor AEG-1, utilizou-se o método de sequenciação por PCR. Neste estudo foram encontradas duas novas mutações C>T (-633) e G>C (-660) no grupo de pacientes. Mas não foi revelada a associação estatisticamente significante entre quaisquer mutações nessa região do promotor AEG-1 com suscetibilidade ao CHC. De acordo com os dados apresentados, podemos dizer que a região reguladora negativa das mutações do promotor AEG-1 não demonstrou relevância significativa com o carcinoma hepatocelular. Recomendamos estudos adicionais sobre a eficácia do promotor AEG-1 no direcionamento terapêutico do CHC.

MiR-22-3p suppresses cell proliferation by targeting AEG-1 in NSCLC / 实用医学杂志

Objective To investigate the expression and effect of miR-22-3p in non-small cell lung cancer (NSCLC). Methods The miR-22-3p expression level in seventy-six NSCLC tissues and para-cancer tissues was detected by qRT-PCR. The relationship between the expression of miR-22-3p and gender,age,tumor size,histolo-gy grade,pathological type and lymph node metastasis was analyzed. The function of miR-22-3p on the prolifera-tion of NSCLC cells was tested by growth curve assay. Target genes of miR-22-3p were predicted by online software Targetscan. Luciferase reporter assay and qRT-PCR was used to certificate the prediction. Results The expression of miR-22-3p was increased in NSCLC tissues than the para-cancer tissues and was correlated to lymph node metas-tasis. Overexpression of miR-22-3p could suppress the proliferation of A549 cells. Astrocyte-Elevated Gene-1(AEG-1) was predicted to be a target of miR-22-3p. MiR-22-3p was revealed to bind to AEG-13′UTR by luciferase report-er assay. Overexpression of miR-22-3p could inhibit the expression of AEG-1 in A549 cells. Suppression of miR-22-3p could increase AEG-1 expression. Conclusion MiR-22-3p could inhibit the proliferation of NSCLC by tar-geting AEG-1.

Expressions and Clinical Significances of AEG-1 and MMP-9 in the Oral Squamous Cell Carcinoma / 现代生物医学进展

Objective:To study the expressions and clinical significances of astrocyte elevated gene-1 (AEG-1) and matrix metal loproteinase-9 (MMP-9) in the oral squamous cell carcinoma (OSCC).Methods:The expressions of AEG-1 and MMP-9 were detected by immunohistochemistry method in 53 cases of OSCC patients and 30 cases of controls with normal oral mucosa tissues.The relation ship of AEG-1 and MMP-9 expressions with the clinicopathologic features were analyzed.Results:The postive expressions of AEG-1 and MMP-9 in the OSCC(66.03 ％,73.58 ％) were signifiacnatly higher than those of the normal oral mucosa tissue (13.33 ％,16.67 ％) (P＜0.01).The expression of AEG-1 was positively correlated with with the expression of MMP-9 (r=0.474,P＜0.01).The high expressions of AEG-1 and MMP-9 were associated with the differentiation degree,lymph node metastasis and clinical stage,but they were not correlated with the gender,age and smoker.Conclusions:The high expression of AEG-1 and MMP-9 were closely related to the occurrence,development and metastasis of OSCC.Therefore,combined detection of AEG-1 and MMP-9 might be valuable for the early diagnosis and prognostic prediction of OSCC.

AEG-1 promotes metastasis of breast cancer MCF-7 cells / 实用肿瘤学杂志

Objective The objective of this study was to observe the effect of AEG-1 gene on the metastasis in breast cancer MCF-7 cells.Methods AEG-1 siRNAs were transfected into MCF-7 cells to silence AEG-1 expression,and negative siRNA was used as a control.Transwell chamber was used to detect the ability of cell migration and invasion of MCF-7 cells.CCK8 assay was used to detect the cell proliferation of MCF-7 cells.At the same time,the effect of VEGF on the angiogenesis was investigated by detecting the changes of lumen formation in HUVEC cells.Results The migration,invasive and proliferative abilities were significantly inhibited in MCF-7 cells transfected with AEG-1 siRNA.Knockdown AEG-1 was significantly decreased the level of VEGF in the supernatant of MCF-7 cells.Knockdown AEG-1 was also significantly inhibited the angiogenesis activity in HUVEC cells.Conclusion Knockdown AEG-1 can significantly inhibit the migration of MCF-7 cells,including cell migration,invasion,proliferation and angiogenesis.These results suggest that AEG-1 plays an important role in the metastasis process of breast cancer and opens up new ideas for future treatment breast cancer.