LncRNA NORAD mediates KMT2D expression by targeting miR-204-5p and affects the growth of gastric cancer.

Background: Long non-coding ribonucleic acids (lncRNAs) are a class of non-coding RNAs implicated in the development of many malignancies, including gastric cancer (GC). In this study, we investigated the functions and molecular mechanisms of non-coding RNA activated by deoxyribonucleic acid damage (NORAD) in GC. Methods: NORAD expression at the messenger RNA levels was determined by quantitative reverse transcriptase (RT)-polymerase chain reaction assays. Cell proliferation, migration, and invasion were detected by Cell Counting Kit-8 assays, in-vivo tumor formation assays, and Transwell assays. Cell-cycle distribution was detected by a flow cytometry analysis. NORAD location was detected by nucleocytoplasmic fractionation assays. The interaction between NORAD and the microRNA-204-5p (miR-204-5p)/Lysine Methyltransferase 2D (KMT2D) axis was verified by dual-luciferase reporter gene assays and RNA binding protein immunoprecipitation (RIP) assays. Western blot was used to study the phosphatase and tensin homolog (PTEN)/phosphoinositide 3-kinases (PI3K)/protein kinase B (AKT) signaling pathway. Results: NORAD was upregulated in the GC tissues and cell lines. The silencing of NORAD repressed cell proliferation and the Growth 2 (G2)/Mitosis (M) cell-cycle transition in GC. NORAD also regulated KMT2D expression by targeting miR-204-5p and mediated PTEN/PI3K/AKT signaling in GC. Conclusions: We found that NORAD acts as an oncogene in GC. Our findings might provide a novel therapeutic target for GC.

Long Intergenic Non-Protein Coding RNA 1089 Suppresses Cell Proliferation and Metastasis in Gastric Cancer by Regulating miRNA-27a-3p/Epithelial-Mesenchymal Transition (EMT) Axis.

AIM: To explore the expression and biological function of long intergenic non-protein coding RNA 1089 (LINC01089) in gastric cancer (GC) progression and its underlying mechanism. METHODS: LINC01089 and microRNA-27a-3p (miR-27a-3p) expressions were detected with the quantitative real-time polymerase chain reaction (RT-qPCR). Cell proliferation, migration and invasion were evaluated by Cell Counting Kit-8 (CCK-8) and Transwell assay. Epithelial-mesenchymal transition (EMT)-related proteins were also measured by Western blot. The relationship between LINC01089 and miR-27a-3p was revealed by a bioinformatics analysis and dual-luciferase reporter assay. RESULTS: LINC01089 was significantly down-regulated in GC tissues, as well as GC cell lines. GC patients with lower LINC01089 expression were more likely to have poor outcomes. Overexpression of LINC01089 significantly suppressed GC cells growth, migration and invasion and forbade the EMT process. LINC01089 was directly targeted at miR-27a-3p. The transfection of miR-27a-3p mimics reversed the inhibitory effects on proliferative and metastatic abilities of GC cells with LINC01089 overexpression. CONCLUSION: LINC01089 inhibits cell proliferation and metastasis in GC by targeting miR-27a-3p/EMT axis, which should be considered as a promising therapeutic target.

Dysregulation of CircRNA_0001946 Contributes to the Proliferation and Metastasis of Colorectal Cancer Cells by Targeting MicroRNA-135a-5p.

This study was aimed to evaluate the potential function of circ-0001946 in the progression of colorectal cancer (CRC) and the related regulatory mechanism. First, the expression levels of circRNA_0001946 and microRNA-135a-5p (miR-135a-5p) in normal and CRC tissues were measured by quantitative real-time polymerase chain reaction (RT-qPCR). In addition, cell proliferation was assessed by the Cell Counting Kit-8 (CCK-8) assay, cell migration and invasion were evaluated by Transwell assays, and the cell cycle patterns were determined by flow cytometry. The relationship between the expression levels of circ_0001946 and miR-135a-5p was determined by dual-luciferase reporter assays. Our data showed that the expression of circ_0001946 was upregulated in CRC tissues, which was negatively correlated with tumor size, histologic grade, lymphatic metastasis, and TMN stage, and patients with circ_0001946 overexpression were more likely to have a poor prognosis. In addition, in vitro experiments showed that silencing circ_0001946 inhibited the epithelial-mesenchymal transition (EMT) pathway and markedly suppressed CRC cell growth, migration, and invasion. Furthermore, we discovered that the transfection of miR-135a-5p mimics could reverse the antitumor effects of circRNA_0001946 downregulation. To summarize, this study revealed that circRNA_0001946 might act as a tumor promoter by activating the miR-135a-5p/EMT axis and may be a promising treatment target for CRC.

Next-Generation Sequencing Analysis of mRNA Profile in Cisplatin-Resistant Gastric Cancer Cell Line SGC7901.

BACKGROUND Cisplatin-resistant gastric cancer (GC) occurs in patients with GC treated with cisplatin-based chemotherapy, which results in disease progression and early recurrence during the treatment. MATERIAL AND METHODS To understand the initiation and developmental mechanism underlying cisplatin-resistant GC, we developed cisplatin-resistant SGC7901 cells (SGC7901/DDP) from the parental cells (SGC7901/S) by continuous exposure to increasing concentrations of cisplatin and subjected these 2 cell lines to RNA sequencing analysis. The data were verified by quantitative polymerase chain reaction and their functional role was evaluated by cell counting kit 8 assay and cell apoptosis and cell cycle flow cytometric analysis. Bioinformatics analysis was performed to classify the differentially-expressed genes (DEGs) involved in the development of cisplatin resistance. RESULTS In comparison with SGC7901/S cells, SGC7901/DDP cells showed a total of 3165 DEGs (2014 upregulated and 1151 downregulated, fold change ≥2, and adjusted P value <0.001). qRT-PCR confirmed the reliability of the RNA sequencing results. Depletion of the top 5 upregulated mRNAs reversed the resistant index, increased apoptotic SGC7901/DDP cells, and arrested the cells at G2/M phase. Gene ontology analysis revealed that the DEGs mainly regulate metabolic process, immune system, locomotion, cell adhesion, cell growth, cell death, cytoskeleton organization, cell binding, signal transducing activity, and antioxidant activity. Kyoto Encyclopedia of Genes and Genomes analysis showed that the DEGs were mainly involved in the PI3K-Akt signaling pathway, Rap1 signaling pathway, proteoglycans in cancer, regulation of actin cytoskeleton, and pathways in cancer. CONCLUSIONS The present study is the first to interrogate mRNAs profiles in human GC cells with cisplatin resistance using RNA sequencing, which may assist in discovering potential therapeutic targets for cisplatin-resistant GC patients.

Successful treatment of posttraumatic phlegmasia cerulea dolens by reconstructing the external iliac vein: a case report.

INTRODUCTION: Phlegmasia cerulea dolens is a rare condition caused by complete venous occlusion leading to impaired arterial flow. To prevent progression to limb gangrene, prompt diagnosis and treatment initiation are paramount. Here we report a rare case of posttraumatic phlegmasia cerulea dolens after ligation of the iliac vein to save the patient's life, with successful treatment by reconstructing the external iliac vein. This is the first report of posttraumatic phlegmasia cerulea dolens induced by iliac vein ligation. CASE PRESENTATION: A 49-year-old Chinese man was admitted to a local hospital for severe knife trauma with massive intraperitoneal bleeding. During exploratory laparotomy, he was diagnosed with traumatic rupture of his left external iliac vein without injury to the iliac artery. The proximal and distal parts of his injured external iliac vein were ligated to control the bleeding and rescue him, but his left leg quickly became severe swollen, cyanotic and pulseless. He was diagnosed with posttraumatic phlegmasia cerulea dolens after being transferred to our university hospital. After a retrievable filter was placed in his inferior vena cava via his right femoral vein, he underwent reopening of his abdomen followed by successful surgical reconstruction of his left iliac vein. He was treated with anticoagulation therapy postoperatively and his signs and symptoms improved markedly. He was discharged in a stable condition, with nearly full resolution of symptoms, 35 days after the operation. CONCLUSIONS: Our case demonstrates that ligation of an injured iliac vein may induce phlegmasia cerulea dolens in a posttraumatic scenario; prompt reconstruction of the iliac vein to restore the venous drainage is an effective treatment for phlegmasia cerulea dolens with impending gangrene.

[Management and prognosis of acute arterial embolism: a multivariable analysis of 346 patients].

OBJECTIVE: To evaluate the management of acute arterial embolism (AAE) and its prognostic factors. METHODS: The clinical data of 346 AAE patients treated at our hospital between January 1998 and October 2008 were retrospectively reviewed. The prognostic factors, including age, gender, extremities, location of embolism, ischemic duration, ischemic categories, and therapeutic methods, postoperative complications were evaluated by multivariate Logistic regression analysis. RESULTS: There were 210 males and 136 females with a mean age of (63 ± 14) years old. Fifty-six patients occurred in the upper extremities and 290 patients in the lower extremities. The causes included cardiogenic embolism (n = 301), vasogenic embolism (n = 33) and unknown origin (n = 12). The duration of ischemia ranged from 1 h to 7 d. Only 44 patients were admitted ≤ 8 h and the remainder > 8 h. The categories of extremity ischemia were level I (n = 17), level IIA (n = 69), level IIB (n = 221) and level III (n = 39). The procedures included embolectomy (n = 280), interventional thrombolysis (n = 19) and conservative treatment (n = 47). Thirteen patients (3.76%) died of complications during the perioperative periods. And 44 (12.72%) underwent amputations and 289 (83.53%) had excellent clinical outcome with extremity salvage. During a 5-year follow-up period, 38 patients had a recurrent embolism. The Logistic regression analysis showed that ischemic duration, ischemic category, therapeutic methods and complications had significant prognostic influences (all P < 0.05). And other factors such as age, gender, extremities and the location of embolism had insignificant influences (all P > 0.05). CONCLUSION: Embolectomy is the first-choice therapy for AAE with an excellent outcome. Ischemic duration, ischemic grading, surgical treatment and complications have significant prognostic influences. Systematic medical treatments, such as effective anticoagulation, are vital in the prevention of recurrent AAE.