ABSTRACT
Abstract Objectives Emerging evidence has demonstrated that LINC01857 exerts a pivotal function in many cancers. However, its function in Pancreatic Ductal Adenocarcinoma (PDAC) still remains unclear. This study was designed to investigate the regulatory character of LINC01857 in PDAC. Methods Bioinformatic tools and databases were used to seek potential miRNAs and mRNAs. Gene expression was evaluated by Reverse Transcription quantitative real-time Polymerase Chain Reaction (RT-qPCR), and western blot was used for protein level detection. A subcellular fraction assay was done to ascertain the location of LINC01857 in PANC-1 and BxPC-3 human pancreatic cancer cells. CCK-8, EdU, wound healing and Transwell assays were performed to inquire into the influence of LINC01857, and SPARC -related Modular Calcium-binding protein-2 (SMOC2) on cell viability, proliferation, migration, and invasion, respectively. The interaction between LINC01857 and its downstream genes was explored by RNA immunoprecipitation and luciferase reporter assays. Results LINC01857 levels were significantly elevated in PDAC. Knockdown of LINC01857 significantly restrained the proliferation, migration, invasion, and Epithelial-Mesenchymal Transition (EMT) process of PDAC cells. MiR-19a-3p was a downstream target of LINC01857, and miR-19a-3p levels were significantly decreased in PDAC cells. In addition, SMOC2 expression had a negative correlation with that of miR-19a-3p, and SMOC2 was a downstream target of miR-19a-3p. Furthermore, SMOC2 upregulation partially abolished the inhibitive influence of LINC01857 downregulation on cell proliferation, migration, invasion, and the EMT process. Conclusion LINC01857 promotes malignant phenotypes of PDAC cells via upregulation of SMOC2 by interacting with miR-19a-3p. HIGHLIGHTS LINC01857 is upregulated in PAAD and promotes malignant cellular behaviors. LINC01857 interacts with miR-19a-3p to regulate SMOC2 expression. LINC01857 promotes malignant cellular phenotypes by upregulating SMOC2.
ABSTRACT
@#AIM: To investigate whether long-chain non-coding RNA(lncRNA)KCNQ1OT1 affects the proliferation, apoptosis and oxidative stress of retinal epithelial cells induced by high glucose(HG)through miR-19a-3p/TSHZ3. <p>METHODS: Cell counting kit 8(CCK-8)was used to detect the cell viability of human retinal epithelial cells ARPE-19 stimulated with 5, 15, 45, 135mmol/L HG. The ARPE-19 cells were divided into NC group, 45mmol/L HG group, si-NC+45mmol/L HG group, si-lncRNA KCNQ1OT1+45mmol/L HG group, miR-NC+45mmol/L HG group, miR-19a-3p mimics+45mmol/L HG group, si-con+45mmol/L HG group, si-TSHZ3+45mmol/L HG group, pcDNA+si-lncRNA KCNQ1OT1+45mmol/L HG group, pcDNA-TSHZ3+si-lncRNA KCNQ1OT1+45mmol/L HG group. CCK-8 was used to detect cell viability, qRT-PCR was used to detect the expressions of lncRNA KCNQ1OT1, miR-19a-3p and TSHZ3 mRNA, Western Blot was used to detect TSHZ3, activation-cysteine-containing aspartate proteolytic enzyme 3(Cleaved-caspase-3), B-cell lymphoma/leukemia-2(Bcl-2)related X protein(Bax)protein expressions, and enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of oxidative stress indicators reactive oxygen species(ROS)and malondialdehyde(MDA). The dual luciferase activity was used to detect the targeted binding between lncRNA KCNQ1OT1 and miR-19a-3p, miR-19a-3p and TSHZ3. <p>RESULTS: 15, 45, 135mmol/L HG inhibited the survival rate of ARPE-19 cells, and the subsequent select the HG concentration 45mmol/L with a cell survival rate of about 50%. 45mmol/L HG increased the expression levels of lncRNA KCNQ1OT1, TSHZ3 mRNA, TSHZ3 protein, the apoptosis rate, Cleaved-caspase-3 and Bax protein expressions, ROS and MDA levels in ARPE-19 cells, and reduced cell survival rate and the expression level of miR-19a-3p(<i>P</i><0.05). Low expression of lncRNA KCNQ1OT1, TSHZ3 or high expression of miR-19a-3p improved the survival rate of ARPE-19 cells induced by HG, and reduced apoptosis rate, Cleaved-caspase-3 and Bax protein expressions, ROS and MDA levels(<i>P</i><0.05). lncRNA KCNQ1OT1 targeted miR-19a-3p, miR-19a-3p targeted TSHZ3, and lncRNA KCNQ1OT13 regulated the expression of TSHZ3 through miR-19a-3p. The effect of lncRNA KCNQ1OT1 low expression on the survival rate, apoptosis and oxidative stress of ARPE-19 cells induced by HG was reversed by the overexpression of TSHZ3.<p>CONCLUSION: The low expression of lncRNA KCNQ1OT13 promotes the proliferation of retinal epithelial cells induced by high glucose, and inhibits their apoptosis and oxidative stress through miR-19a-3p/TSHZ3.
ABSTRACT
@# Objective: To explore the mechanism of miR-19a-3p regulating cell adhesion molecule 2 (CADM2) to inhibit the proliferation and metastasis of renal carcinoma cells via the AKT signaling pathway. Methods: A total of 42 patients with renal cancer admitted to Department of Nephrology, the First Affiliated Hospital of Suzhou University from April 2012 to November 2017 were enrolled to collect samples of surgically resected renal carcinoma tissues and paracancerous tissues. Expression of miR-19a-3p was detected in renal carcinoma tissues and 4 types of renal carcinoma cell lines such as 786-O by quantitative Real-time polymerase chain reaction (qPCR). The effects of miR-19a-3p knockdown on proliferation, invasion and epithelial mesenchymal transition (EMT) of renal carcinoma 786-O cells were evaluated by CCK-8 assay, Transwell assay and immunofluorescence, respectively. Subsequently, dual luciferase reporter assay was used to verify whether CADM2 was a target gene of miR-19a-3p. Furthermore, Wb was applied to detect the regulatory effect of miR-19a-3p onAKT signaling pathway through CADM2. Results: miR-19a-3p expression was significantly up-regulated in renal carcinoma tissues and cell lines (all P<0.01). Knockdown of miR-19a-3p could inhibit proliferation, invasion and EMT process of 786-O cells; furthermore, the results indicated that CADM2 was a direct target of miR-19a-3p and its expression was down-regulated (P <0.05 or P<0.01). Additionally, knockdown of miR-19a-3p obviously suppressed proliferation, migration and EMT process of 786-O cells via up-regulating CADM2 and blocking AKT pathway (all P<0.05 or P<0.01), thus alleviating the occurrence and development of renal carcinoma. Conclusion: The study demonstrates that miR-19a-3p has a high expression level in renal carcinoma tissues; knockdown of miR-19a-3p could significantly inhibit the proliferation, migration and EMT process of renal carcinoma tissues, and its mechanism may be associated with miR-19a-3p/CADM2/AKT axis.
ABSTRACT
MTUS1 (microtubule-associated tumor suppressor 1) has been identified that can function as a tumor suppressor gene in many malignant tumors. However, the function and mechanisms underlying the regulation of MTUS1 are unclear. In the present study, we reported that miR-19a and miR-19b (miR-19a/b) promote proliferation and migration of lung cancer cells by targeting MTUS1. First, MTUS1 was proved to function as a tumor suppressor in lung cancer and was linked to cell proliferation and migration promotion. Second, an inverse correlation between miR-19a/b expression and MTUS1 mRNA/protein expression was noted in human lung cancer tissues. Third, MTUS1 was appraised as a direct target of miR-19a/b by bioinformatics analysis. Fourth, direct MTUS1 regulation by miR-19a/b in lung cancer cells was experimentally affirmed by cell transfection assay and luciferase reporter assay. Finally, miR-19a/b were shown to cooperatively repress MTUS1 expression and synergistically regulate MTUS1 expression to promote lung cancer cell proliferation and migration. In conclusion, our findings have provided the first clues regarding the roles of miR-19a/b, which appear to function as oncomirs in lung cancer by downregulating MTUS1.
Subject(s)
Female , Humans , Male , A549 Cells , Cell Movement , Cell Proliferation , Gene Expression Regulation, Neoplastic , Lung Neoplasms , Genetics , Metabolism , Pathology , MicroRNAs , Genetics , Metabolism , RNA, Neoplasm , Genetics , Metabolism , Tumor Suppressor Proteins , GeneticsABSTRACT
Objective To investigate the effect of knocking down miR-19a on proliferation and apoptosis of human glioma cell line U251 in vitro.Methods U251 cells were cultured routinely.MiR-19a inhibitor was transfected into U251 cells by Lipofectamine 2000.At the same time,the negative control group and blank control group were established.The expression level of miR-19a was detected by RT-PCR.and cell proliferation was analyzed by CCK-8 assay.The changes of cell apoptosis and cycle were monitored by flow cytometry.Results Compared with the negative control group and blank control group,qRT-PCR showed that the expression level of miR-19a was significantly reduced after transfection (F=124.72,P ＜ 0.01).CCK-8 revealed that proliferation ability of miR-19a inhibitor group was significantly suppressed.The cell survival rate in miR-19a inhibitor group after 48 h was (48.27-±8.23) ％,compared with the blank control group (100.00 ％),the difference was statistically significant (t =12.45,P ＜ 0.01).After low-expression of m iR-19a,the G0/G1 phase cells were increased and S phase cells were decreased.The low-expression of miR-19a could induce cell apoptosis.Conclusions Low-expression of miR-19a can inhibit cell proliferation,block G1/S transition and induce apoptosis in human glioma cell line U251.miR-19a may serve as an attractive target of gene therapy for glioblastoma.