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SUMMARY: Overexpression of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in various tumor tissues and cell lines was found to promote tumor cell proliferation, migration, and invasion. However, the role of MALAT1 in gastric cancer (GC) is still unclear. We aimed to investigate the correlation between long-chain non-coding RNAs (lncRNAs), MALAT1, MicroRNAs (miRNA) and vascular endothelial growth factor A (VEGFA) in gastric cancer and to disclose underlying mechanism. The correlation between MALAT1 levels and clinical features was analyzed by bioinformatics data and human samples. The expression of MALAT1 was down regulated in AGS cells to detect the cell proliferation, migration, and invasion characteristics, as well as the effects on signal pathways. Furthermore, we validated the role of MALAT1/miR-330-3p axis in GC by dual luciferase reporter gene assays. Expression of MALAT1 was higher in cancer tissues than in para-cancerous tissues. The high MALAT1 level predicted malignancy and worse prognosis. Down-regulation of MALAT1 expression in AGS cells inhibited cell proliferation, migration, and invasion by targeting VEGFA. By dual luciferase reporter gene assay and miR-330-3p inhibitor treatment, we demonstrate that MALAT1 sponged miR-330-3p in GC, leading to VEGFA upregulation and activation of the mTOR signaling pathway. The MALAT1/miR-330-3p axis regulates VEGFA through the mTOR signaling pathway and promotes the growth and metastasis of gastric cancer.
Se descubrió que la sobreexpresión del transcrito 1 de adenocarcinoma de pulmón asociado a metástasis (MALAT1) en varios tejidos tumorales y líneas celulares promueve la proliferación, migración e invasión de células tumorales. Sin embargo, el papel de MALAT1 en el cáncer gástrico (CG) aún no está claro. Nuestro objetivo fue investigar la correlación entre los ARN no codificantes de cadena larga (lncRNA), MALAT1, los microARN (miARN) y el factor de crecimiento endotelial vascular A (VEGFA) en el cáncer gástrico y revelar el mecanismo subyacente. La correlación entre los niveles de MALAT1 y las características clínicas se analizó mediante datos bioinformáticos y muestras humanas. La expresión de MALAT1 se reguló negativamente en las células AGS para detectar las características de proliferación, migración e invasión celular, así como los efectos sobre las vías de señales. Además, validamos el papel del eje MALAT1/miR- 330-3p en GC mediante ensayos de genes indicadores de luciferasa dual. La expresión de MALAT1 fue mayor en tejidos cancerosos que en tejidos paracancerosos. El alto nivel de MALAT1 predijo malignidad y peor pronóstico. La regulación negativa de la expresión de MALAT1 en células AGS inhibió la proliferación, migración e invasión celular al apuntar a VEGFA. Mediante un ensayo de gen indicador de luciferasa dual y un tratamiento con inhibidor de miR-330-3p, demostramos que MALAT1 esponjaba miR-330-3p en GC, lo que lleva a la regulación positiva de VEGFA y la activación de la vía de señalización mTOR. El eje MALAT1/miR-330-3p regula VEGFA a través de la vía de señalización mTOR y promueve el crecimiento y la metástasis del cáncer gástrico.
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AIM: To investigate the effect of lncRNA MALAT1 on the proliferation, migration and angiogenesis of retinal vascular endothelial cells and its molecular mechanism.METHODS: The expression levels of lncRNA MALAT1 in plasma of normal control group, diabetic without retinopathy group and diabetic retinopathy group were detected by qPCR and the effect of glucose culture on the expression levels of lncRNA MALAT1 were detected by qPCR too. The expression level of miR-124-3p was detected by qRT-PCR; Western blotting was used to detect the expression level of SOX7; The targeting relationship between lncRNA MALAT1 and miR-124-3p, miR-124-3p and SOX7 were detected by the dual-luciferase reporter system; CCK-8 assay was used to detect cell proliferation activity; Transwell assay was used to detect the migration ability of cells; Angiogenesis of hRMECs cells was measured by in vitro tube formation assay.RESULTS:The expression level of lncRNA MALAT1 in plasma of diabetic retinopathy patients was significantly higher than that of diabetic without retinopathy group and normal control group(P<0.001). In vitro glucose culture significantly promoted the expression of lncRNA MALAT1 in hRMECs cells, as well as the proliferation, migration and angiogenesis of hRMECs cells(all P<0.05). Knockdown of lncRNA MALAT1 significantly inhibited the proliferation, migration and tubule formation of hRMECs cells(all P<0.05). Dual-luciferase reporter gene assay showed that lncRNA MALAT1 targeted with miR-124-3p, and miR-124-3p targeted with SOX7. Overexpression of miR-124-3p significantly inhibited the proliferation, migration and tubule formation of hRMECs cells(all P<0.05). Overexpression of lncRNA MALAT1+miR-124-3p, miR-124-3p+SOX7, and knockdown of lncRNA MALAT1+overexpression of SOX7 all significantly eliminated the inhibitory effect of hRMECs cells(all P<0.05).CONCLUSION: lncRNA MALAT1 promote the proliferation, migration and angiogenesis of retinal endothelial cells in diabetic retinopathy by down-regulating the negative regulation of miR-124-3p on SOX7. Therefore, abnormal upregulation of lncRNA MALAT1 in patients with diabetic retinopathy is a potential biomarker.
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We aimed to investigate the association of long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1 (lnc-MALAT1) with acute ischemic stroke (AIS), and its association with disease severity, inflammation, and recurrence-free survival (RFS) in AIS patients. One hundred and twenty AIS patients and 120 controls were recruited. Venous blood samples from AIS patients (within 24 h after symptoms onset) and controls (at entry to study) were collected to detect plasma lnc-MALAT1 expression by real-time quantitative polymerase chain reaction. AIS severity was assessed by the National Institutes of Health Stroke Scale (NIHSS) score. Plasma concentrations of inflammation factors (including C-reactive protein (CRP), tumor necrosis factor α (TNF-α), interleukin (IL)-6, IL-8, IL-10, IL-17, and IL-22) were measured and RFS was calculated. lnc-MALAT1 expression was decreased in AIS patients compared to controls, and it had a close correlation with AIS (AUC=0.791, 95% CI: 0.735-0.846). For disease condition, lnc-MALAT1 expression negatively correlated with NIHSS score and pro-inflammatory factor expression (including CRP, TNF-α, IL-6, IL-8, and IL-22), while it positively correlated with anti-inflammatory factor IL-10 expression. Furthermore, lnc-MALAT1 expression was elevated in AIS patients with diabetes. For prognosis, no statistical correlation of lnc-MALAT1 expression with RFS was found, while a trend for longer RFS was observed in patients with lnc-MALAT1 high expression compared to those with lnc-MALAT1 low expression.
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Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Idoso , Isquemia Encefálica/diagnóstico , Acidente Vascular Cerebral/diagnóstico , RNA Longo não Codificante/genética , AVC Isquêmico , InflamaçãoRESUMO
@#To investigate whether lncRNA MALAT1 affects the migration and proliferation of breast cancer cells through the regulation with histone methyltransferase SMYD3, the endogenous MALAT1 in the MCF-7 and MDA-MB-231 breast cancer cells were knocked down by siRNA, and then the migration and proliferation of cells were detected by wound healing migration and MTT assay. The effects of si-MALAT1 on the mRNA and protein levels of miRNA-124, SMYD3 and its downstream genes were detected by Real time PCR and Western blot. The results showed that siRNA-targeted knockdown of MALAT1 reduced the migration and proliferation of breast cancer cells, and inhibited the transcriptional expression of SMYD3 and its downstream genes, including N-cadherin, MYL9, MMP9 and CYR61, and up-regulated miR-124. Overexpression of miR-124 reduced the expression of SMYD3 in breast cancer cells, and knockdown of MALAT1 attenuated the promotion of SMYD3 protein expression by miR-124 inhibitors. In addition, SMYD3 overexpression activated MALAT1 transcription, whereas siRNA interference with SMYD3 downregulated MALAT1. These results indicate that LncRNA MALAT1 acted as a competing endogenous RNA(ceRNA)of miR-124 to regulate expression of SMYD3 in breast cancer cells, and SMYD3 can activate the transcription of MALAT1, which will affect the proliferation and migration of breast cancer cells.
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To study the effect of small interfering RNA targeting metastasis-associated lung adenocarcinoma transcript1 (si-MALAT1) combining with curcumin on the invasion and migration abilities of human colon cancer SW480 cells, and to explore the involved molecular mechanism. The recombinant lentiviral vector expressing si-MALAT1 was constructed, and its titer was determined by gradient dilution method. The colon cancer SW480 cells with stable expression of si-MALAT1 was established, followed by treatment with curcumin at different concentrations. The effect of curcumin or si-MALAT1 alone and the combination of the two on the cell activity was detected by MTT assay. The cell invasion and migration abilities were detected by transwell and scratch-wound assay. The relative expression level of MALAT1 was detected by RT-qPCR. The protein expression was determined by Western blot analysis. The IC50 of curcumin alone was 77.69 mmol/L, which was 51.17 mol/L when combined with curcumin and random sequence. The IC50 of curcumin was 30.02 mmol/L when combined with si-MALAT1. The increased susceptibility multiples was 2.58. The wound healing rates were 30.9% and 67.5% after treatment with si-MALAT1 combined with curcumin for 24 hrs and 48 hrs, respectively. The numbers of invasion cells were 200±12, 162±13, 66±8, 53±4 and 16±3 after treatment with si-MALAT1 combined with curcumin for 48 hrs. The relative expression level of lncRNA-MALAT1 in the curcumin group was 68%, and the relative expression level of lncRNA-MALAT1 in si-MALAT1group was 56%, and that for the combination treatment group was about 21%. The protein expression levels of β- catenin, c-myc and cyclinD1 were significantly down-regulated upon treatment with certain concentration of si-MALAT1 alone or combined with curcumin.si-MALAT1 could significantly inhibit the invasion and migration of SW480 cells by enhancing the sensitivity of SW480 cells to curcumin. The mechanism involved mignt be related to the down-regulation of β-catenin, c-myc and cyclinD1 proteins.
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Inibição de Migração Celular/efeitos dos fármacos , Neoplasias do Colo , Curcumina/farmacologia , Neoplasias/prevenção & controle , RNA , RNA Interferente Pequeno/efeitos dos fármacosRESUMO
Objective:To investigate the relationship between LncRNA MALAT-1 and miR-205 in non-small cell lung cancer and the mechanism of biological behavior in lung cancer cells.Methods: The expression of LncRNA MALAT-1 in different non-small cell lung cancer was detected by qPCR.The interaction between MALAT-1 and miR-205 was detected by double luciferase reporter gene.Transwell invasion assay and scratch test were used to detect the changes of invasion and migration abilities of lung cancer cells after silencing MALAT-1 and the recovery the abilities after inhibition of miR-205 expression.The tumor volume and quality of lung cancer cells were detected by subcutaneous tumor formation in nude mice after silencing MALAT-1.Results:Compared with other lung cancer cell lines,the expression of MALAT-1 was the highest and the expression level of miR-205 was the lowest in A549 cells.The double luciferase assay confirmed that MALAT-1 could specifically bind to 3′UTR of miR-205,and could regulate the expression and activity of miR-205.Inhibition of MALAT-1 expression could reduce the migration and invasion of lung cancer cells;while inhibiting the expression of miR-205 level,the migration and invasion of lung cancer cells could recovery.The tumor volume and quality of lung cancer cells were reduced after silencing MALAT-1 in subcutaneous tumorigenesis of nude mice.Conclusion: MALAT-1 can regulate the expression of miR-205 and affect the invasion and migration of lung cancer cell line A549.