Prognostic value and mechanism of long non-coding RNA DLEU1 in osteosarcoma / 中国骨伤

OBJECTIVE@#To investigate the prognostic value and mechanism of long non-coding RNA DLEU1(LncRNA DLEU1) in osteosarcoma.@*METHODS@#The tissue samples and clinical data of 86 patients with osteosarcoma treated by orthopaedic surgery in our hospital from January 2012 to December 2014 were retrospectively collected. The expression of LncRNA DLEU1 in pathological tissues was detected by qRT-PCR, then the patients were divided into high and low expression of LncRNA DLEU1 groups. Osteosarcoma cell line HOS was divided into two groups, down-regulated expression group (si-DLEU1 group) and negative control group (si-NC group). LncRNA DLEU1 siRNA and negative control sequence were transfected by Lipofectamine 3000. Chi-square test was used to analyze the relationship between the expression of LncRNA DLEU1 and the clinicopathological factors of osteosarcoma. Kaplan-Meier method was used to compare the difference of the overall survival rate of osteosarcoma patients between the high and low expression groups of LncRNA DLEU1. The risk factors affecting the overall survival rate of osteosarcoma were analyzed by single factor and multifactor analysis. The number of invasive cells in the two groups was determined and compared by Transwell assay.@*RESULTS@#The expression of LncRNA DLEU1 in osteosarcoma tissue was higher than that in adjacent tissues (P<0.001). The expression of LncRNA DLEU1 in human osteosarcoma cell lines (MG-63, U-2 OS, and HOS) was significantly higher than that in human osteoblast line hFOB 1.19 (P<0.001). The expression of LncRNA DLEU1 was significantly correlated with Enneking stage (P<0.001), distant metastasis (P=0.016), and histological grade (P=0.028). The 1-year overall survival rate of the LncRNA DLEU1 high expression group was significantly higher than that of the low expression group (90.7% vs 60.5%, P<0.001). The 5-year overall survival rate of the LncRNA DLEU1 high expression group was significantly higher than that of the low expression group (32.6% vs 11.6%, P<0.001). Univariate analysis showed that Enneking stage (P<0.001), tumor size (P=0.043), distant metastasis (P<0.001), histological grade (P<0.001), and expression of LncRNA DLEU1 (P<0.001) were risk factors for overall survival of osteosarcoma patients. Multivariate analysis showed that high expression of LncRNA DLEU1 [HR=1.948, 95% CI(1.141, 3.641), P=0.012] and distant metastasis[HR=4.108, 95% CI(2.169, 7.780), P<0.001] were independent risk factors for overall survival of osteosarcoma patients. The number of invasive cells in si-DLEU1 group was significantly lesser than that in si-NC group(139±13 vs 357±31, P<0.001).@*CONCLUSION@#High expression of LncRNA DLEU1 is a molecular marker affecting the prognosis of osteosarcoma patients. Downregulation of LncRNA DLEU1 can inhibit the invasion of osteosarcoma cells.

Expression of LncRNA DLEU1 in Esophageal Squamous Cell Carcinoma and Its Effect on Proliferation and Migration of ESCC Cells / 肿瘤防治研究

Objective To investigate the expression of LncRNA DLEU1 in ESCC tissues, and its effect on the proliferation and migration of ESCC cells. Methods We collected 58 cases of ESCC tissues and corresponding para-cancerous tissues. RT-qPCR was used to detect the relative expression levels of DLEU1 in ESCC tissues and cells. Log-rank test was used to analyze the relation between the expression of DLEU1 and clinicopathological features. Kaplan-Meier analysis was used to investigate the correlation between the expression of DLEU1 and the survival of ESCC patients. Multivariate Cox regression model was used to evaluate the effect of DLEU1 on the prognosis of ESCC patients. Effects of DLEU1 on the proliferation and migration of Eca9706 cells were evaluated by CCK-8 and wound healing assays, respectively. Results DLEU1 was highly expressed in ESCC tissues (P < 0.01) and significantly correlated with tumor size, TNM stage and lymph node metastasis (all P < 0.05). High expression of DLEU1 was negatively correlated with poor prognosis of ESCC patients (P < 0.01), and DLEU1 was also an independent prognostic risk factor (P < 0.05). Moreover, knockdown of DLEU1 significantly inhibited the proliferation and migration of Eca9706 cells, compared with the control group (P < 0.01). Conclusion DLEU1 is highly expressed in ESCC tissues. The expression of DLEU1 is an independent risk factor for the prognosis of ESCC patients and promotes ESCC cell proliferation and migration.

Expression of lncRNA DLEU1 in non-small cell lung cancer and its effect on metastasis / 中国肿瘤临床

Objective: To investigate the expression pattern and clinical significance of long non-coding RNA deleted in lymphocytic leu-kemia 1 (DLEU1) in non-small cell lung cancer (NSCLC), and to further evaluate its effect on tumor metastasis. Methods: Paired cancer and adjacent tissues were obtained from 42 patients with NSCLC that underwent surgical resection of cancer in The Third Xiangya Hos-pital of Central South University from January 2008 to December 2012, at the Department of Cardiothoracic Surgery. The expression features of DLEU1 in NSCLC tissue were detected by real-time quantitative polymerase chain reaction assay. Statistical methods were used to analyze the relationship between the expression of DLEU1 and the clinicopathological features and the survival time of pa-tients with NSCLC. Effects of DLEU1 on migration and invasion of NSCLC cells were evaluated by in vitro wound healing and cell inva-sion assays, respectively. Western blot was performed to investigate the protein levels of E-cadherin, N-cadherin, and Vimentin, which are the biomarkers of epithelial to mesenchymal transition (EMT). Results: Of the 42 NSCLC samples, DLEU1 expression was up-regulat-ed in 35 samples (83.33%) and down-regulated in 7 samples (16.67%). The expression level of DLEU1 in NSCLC tissue was 2.11 times that in the adjacent tissues (P<0.05). The expression of DLEU1 in four NSCLC cell lines (A549, H1299, SPCA1, and H358) was higher than that in normal lung 16HBE epithelial cells, and of these, A549 cells had the highest DLEU1 expression (P<0.05). Highly expressed DLEU1 was positively correlated with lymph-node metastasis (P<0.05), but was not significantly associated with other parameters, in-cluding patient gender, age, smoking history, primary tumor size, histological grade, and the TNM stage. The survival time of patients with high DLEU1 expression was significantly shorter than that of patients with low DLEU1 expression (P<0.05). In vitro interference with DLEU1 expression significantly inhibited the migration and invasion of A549 and SPCA1 cells compared with that of the control group (P<0.05). Upon interference with DLEU1 expression , protein levels of E-cadherin increased, whereas those of N-cadherin and Vi-mentin decreased in A549 cells in comparison with the control group. Conclusions: The expression of DLEU1 is up-regulated in NSCLC tissues and cell lines, correlates with lymph-node metastasis and survival times in patients with NSCLC, and promotes tumor cell migra-tion and invasion by regulating EMT, suggesting that DLEU1 may be a potential therapeutic target for NSCLC.

Detection of chromosome 13 (13q14) deletion among Sudanese patients with multiple myeloma using a molecular genetics fluorescent in situ hybridization technique (FISH)

Multiple myeloma (MM) is a neoplastic plasma cell dyscrasia with an incidence of 4-4.5 per 100,000 population per year. It is regarded as the second most prevalent blood cancer (10%) after non-Hodgkin lymphoma. The objective of this study was to investigate the mutational change in chromosome 13 (13q14) among Sudanese MM patients and to identify the association between extent of plasma cell infiltration in the bone marrow, albumin level and deletion of 13q14 by an analytical case control study. Materials and Methods: 15 patients were enrolled in the study. 11 bone marrow samples were collected from MM patients at different stages of the disease and 4 samples were from patients with conditions other than MM as control. Plasma cells were counted from bone marrow smears and fluorescence-in-situ hybridization (FISH) was performed using Fluorophore labeled DLEU1 (13q14) LSI (local specific identifier) probe designed as a dual-colour assay to detect deletion at 13q14. Heparanized sample was taken for estimation of serum albumin in all patients. Results: 13q14 deletion was detected in 6 (54.5%) MM patients while one (9.1%) patient showed monosomy. All relapsed MM (27.3%) had 13q14 deletion. Surprisingly almost all patients studied had normal albumin level. The study could not show whether the deletion is implicated in the pathogenesis of multiple myeloma.