Prognostic thirteen-long non-coding RNAs (IncRNAs) could improve the survival prediction of gastric cancer / El pronóstico de 13 largos RNA no codificantes (IncRNA) podría mejorar la predicción de supervivencia del cáncer gástrico

OBJECTIVE: Accumulating evidence has demonstrated that long non-coding RNAs (lncRNAs) play important regulatory roles in the tumorigenesis and progression of gastric cancer (GC). The aim of this study was to construct the prognostic predictive model of lncRNAs signature and improve the survival prediction of GC. PATIENTS AND METHODS: The expression profiling of lncRNAs in large GC cohorts was performed from The Cancer Genome Atlas (TCGA) databases using the lncRNAs-mining approach, including training data set (N=160) and testing data set (N=159). A 13-lncRNAs signature significantly associated with overall survival (OS) in the training data set was selected. The prognostic value of this 13-lncRNAs signature was then confirmed in the test validation set and the entire validation set, respectively. RESULTS: Based on lncRNA expression profiling of 319 patients with stomach adenocarcinoma (STAD), prognostic 13-lncRNAs signature was found to be significantly associated with the prognosis of GC. Compared to patients with low-risk scores, patients with high-risk scores had a significantly shorter survival time. Moreover, functional enrichment analysis indicated that this 13-lncRNAs signature was potentially involved in multiple biological processes, such as DNA replication and cell cycle signaling pathway. CONCLUSIONS: The prognostic model of the 13-lncRNAs signature established by our study could improve the survival prediction of GC to a greater extent

OBJETIVO: Las pruebas acumuladas demostraron que los ARN no codificantes de larga duración (ARNlC) desempeñaban los importantes papeles reguladores en la tumorigénesis y la progresión del cáncer gástrico (CG). El objetivo de este estudio fue construir el modelo predictivo de pronóstico de la firma de los lncRNA y mejorar la predicción de supervivencia del GC. PACIENTES Y MÉTODOS: El perfil de expresión de los lncARN en grandes cohortes de GC se realizó a partir de las bases de datos del Atlas del Genoma del Cáncer (TCGA) utilizando el enfoque de minería de lncARN, incluyendo el conjunto de datos de entrenamiento (N=160) y el conjunto de datos de pruebas (N=159). Se eligió la firma de 13 lncARN significativamente asociada con la supervivencia general (OS) en la serie de capacitación. El valor pronóstico de esta firma de 13-lncARN se confirmó luego en la serie de validación de pruebas y en toda la serie de validación, respectivamente. RESULTADOS: Basado en el perfil de expresión de lncRNA de 319 pacientes con adenocarcinoma de estómago (STAD), se encontró que la firma de 13-lncRNA de pronóstico estaba significativamente asociada con el pronóstico de GC. En comparación con los pacientes con puntuaciones de bajo riesgo, los pacientes con puntuaciones de alto riesgo tuvieron un tiempo de supervivencia significativamente más corto. Además, el análisis de enriquecimiento funcional indicó que esta firma de 13-lncARN estaba potencialmente involucrada en múltiples procesos biológicos, como la replicación del ADN y la vía de señalización del ciclo celular. CONCLUSIONES: El modelo de pronóstico de la firma de 13-lncARN establecido por nuestro estudio podría mejorar mejor la predicción de supervivencia del GC

AICAR and Decitabine Enhance the Sensitivity of K562 Cells to Imatinib by Promoting Mitochondrial Activity.

Although the advent of tyrosine kinase inhibitors (TKIs) has dramatically improved the survival of patients with chronic myeloid leukaemia (CML), acquired drug resistance and TKI-insensitive leukaemic stem cells (LSCs) remain major obstacles to a CML cure. In recent years, the reprogramming of mitochondrial metabolism has emerged as a hallmark of cancers, including CML, and in turn may be exploited for therapeutic purposes. Here, we investigated the effects of several drugs on the mitochondrial function of the CML cell line K562 and found that 5-aminoimidazole-4-carboxamide ribotide (AICAR) and decitabine could effectively increase the ATP content and mitochondrial biogenesis. In addition, these two drugs induced cell cycle arrest and a decrease in colony-forming capacity and promoted K562 cell differentiation. Moreover, we demonstrated that treatment with AICAR or decitabine enhanced the sensitivity of K562 cells to imatinib, as evidenced by a combination treatment assay. Altogether, our findings indicate that TKIs combined with mitochondrial regulation may provide a therapeutic strategy for the treatment of CML.

Prognostic thirteen-long non-coding RNAs (IncRNAs) could improve the survival prediction of gastric cancer.

OBJECTIVE: Accumulating evidence has demonstrated that long non-coding RNAs (lncRNAs) play important regulatory roles in the tumorigenesis and progression of gastric cancer (GC). The aim of this study was to construct the prognostic predictive model of lncRNAs signature and improve the survival prediction of GC. PATIENTS AND METHODS: The expression profiling of lncRNAs in large GC cohorts was performed from The Cancer Genome Atlas (TCGA) databases using the lncRNAs-mining approach, including training data set (N=160) and testing data set (N=159). A 13-lncRNAs signature significantly associated with overall survival (OS) in the training data set was selected. The prognostic value of this 13-lncRNAs signature was then confirmed in the test validation set and the entire validation set, respectively. RESULTS: Based on lncRNA expression profiling of 319 patients with stomach adenocarcinoma (STAD), prognostic 13-lncRNAs signature was found to be significantly associated with the prognosis of GC. Compared to patients with low-risk scores, patients with high-risk scores had a significantly shorter survival time. Moreover, functional enrichment analysis indicated that this 13-lncRNAs signature was potentially involved in multiple biological processes, such as DNA replication and cell cycle signaling pathway. CONCLUSIONS: The prognostic model of the 13-lncRNAs signature established by our study could improve the survival prediction of GC to a greater extent.

Monitoring and Analysis of Chinese Chronic Myeloid Leukemia Patients Who Have Stopped Tyrosine Kinase Inhibitor Therapy.

Discontinuation of tyrosine kinase inhibitor (TKI) therapy after achieving a persistent deep molecular response (DMR) is an urgently needed treatment goal for chronic myeloid leukemia (CML) patients and has been included in the National Comprehensive Cancer Network (NCCN) guidelines (version 2.2017) for CML. Indeed, various studies have confirmed the feasibility of discontinuing TKI therapy. In this study, we analyzed data from 45 CML patients who had discontinued TKI therapy. Univariate analysis was performed to predict factors that were potentially related to treatment-free remission (TFR) and identify the differences between early relapse and late relapse. Out of the 45 patients, 20 exhibited molecular relapse after a median follow-up of 18 months (range, 1-54 months), and the estimated TFR at 24 months was 40%. The univariate analysis revealed that a high Sokal score and interruptions or dose reductions during TKI treatment were the only baseline factors associated with poor outcomes. Our results indicate that TKI discontinuation could be successfully put into practice in China.

Anti-PD-1 therapy for clinical treatment of lymphoma: a single-arm meta-analysis.

Anti-PD1 antibodies exhibit satisfactory efficacy in treating certain types of lymphoma. We conducted this meta-analysis to explore subtypes benefiting from this treatment and the best anti-PD1 therapeutic modalities. METHODS: A quantitative meta-analysis was performed via a systematic search in PubMed, Web of Science, and The Cochrane Library. The pooled overall response rate (ORR), progression-free survival (PFS), complete remission rate (CRR), overall survival (OS) and adverse events (AEs) were calculated and compared. Data were analyzed using a random-effects meta-analysis to determine risk ratios. Heterogeneity across studies was analyzed using Q and I2 statistics. RESULTS: Thirteen articles were selected, and 9 studies were included in the meta-analysis. There was evidence of significant heterogeneity among the studies. According to PD-L1 expression subgroup analysis, the PD-L1-positive group exhibited significantly better outcomes than the PD-L1-negative group (Z=5.481, p=0.000), with pooled ORRs of 0.74 (95% CI: 0.67-0.81) and 0.2 (95% CI: 0.11-0.3), respectively. For PD-L1-positive and PD-L1-negative patients, the pooled CRRs, PFS and OS were 0.21 (95% CI: 0.14-0.29), 0.76 (95% CI: 0.71-0.81) and 1.0 (95% CI: 0.98-1.0) and 0.05 (95% CI: 0.01-0.11), 0.20 (95% CI: 0.09-0.39) and 0.64 (95% CI: 0.45-0.80), respectively; differences were all statistically significant (Z=2.248, p=0.025; Z=3.555, p=0.000; and Z=3.039, p=0.002, respectively). The pooled incidence of treatment-related all-grade AEs and grade-3/4 AEs was 0.84 (95% CI: 0.75-0.92) and 0.21 (95% CI: 0.15-0.29), respectively. CONCLUSION: Patients with PD-L1 overexpression in relapsed or refractory lymphoma benefited more from anti-PD-1 therapy. Moreover, treatment with approved PD-1 inhibitors was well tolerated.

BCR-ABL1-positive microvesicles malignantly transform human bone marrow mesenchymal stem cells in vitro.

The intercellular communication between leukemia cells and bone marrow mesenchymal stem cells (BM-MSCs) plays more important role in chronic myeloid leukemia (CML) than we previously understood. Recently, we found that microvesicles released from human leukemia cell line K562 (K562-MVs) containing BCR-ABL1 mRNA malignantly transformed normal hematopoietic transplants. Here, we investigated whether K562-MVs contribute to the transformation of human bone marrow mesenchymal stem cells (BM-MSCs). We showed that K562-MVs could be integrated into co-cultured normal BM-MSCs and dose-dependently enhanced the proliferation of BM-MSCs. Meanwhile, K562-MVs (400 ng/mL) significantly increased the expression of BCR-ABL1 in these BM-MSCs, accompanied by the enhanced secretion of TGF-ß1. These BM-MSCs in turn could trigger the TGF-ß1-dependent proliferation of K562 cells. Moreover, we confirmed the presence of BCR-ABL1 in circulating MVs from 11 CML patients. Compared to the normal BM-MSCs, the BM-MSCs from CML patients more effectively increased the BCR-ABL1 expression and TGF-ß1 secretion in K562 cells as well as the proliferation of K562 cells. Our findings enrich the mechanisms involved in the interaction between leukemia cells and BM-MSCs and provide novel ways to monitor minimal residual disease and worthwhile approaches to treat CML.

Detection of cytokine expression patterns in the peripheral blood of patients with acute leukemia by antibody microarray analysis.

The cytokines of acute leukemia (AL) patients have certain expression patterns, forming a complex network involved in diagnosis, progression, and prognosis. We collected the serum of different AL patients before and after complete remission (CR) for detection of cytokines by using an antibody chip. The expression patterns of cytokines were determined by using bioinformatics computational analysis. The results showed that there were significant differences in the cytokine expression patterns between AL patients and normal controls, as well as between acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL). In confirmatory test, ELISA revealed the expression of uPAR in AL. Moreover, the bioinformatic analysis showed that the differentially expressed cytokines among the AL groups were involved in different biological behaviors and were closely related with the development of the disease. It was concluded that the cytokine expression pattern of AL patients is significantly different from that of healthy volunteers. Also, differences of cytokine expression patterns exist between AML and ALL, and between before and after CR in the same subtype of AL, which holds important clinical significance for revealing disease progression.

[Exogenous VEGF promotes hematopoietic stem cell mobilization].

This study was aimed to investigate the effect of exogenous VEGF on hematopoietic stem cell mobilization and immune system. The C57BL/6J mice were randomly divided into the normal control group, VEGF short-term group (5 d) and VEGF long-term group (27 d). Mice in the experimental group were injected ip with VEGF (100 ng/d); mice in control group were injected ip with PBS. The white blood cell (WBC) count and the ratio of lymphocyte in the peripheral blood at different time point were assayed by hemacytometer. The percentage of hematopoietic stem cell (HSC), lymphocyte subgroup, regulatory T cell (Treg), myeloid-derived suppressor cells (MDSC) in the peripheral blood and spleen of different groups were detected by flow cytometry. The morphological changes of spleen and spleen index of mice in the control and long-term group were observed by microscopy. The results showed that the absolute number of WBC in the peripheral blood of mice significantly increased after injection of VEGF, and the peak value was at day 3. The percentage of Lin(-)Sca-1(+)CD117(+) cells in the peripheral blood and spleen of the long-term group were significantly higher than that in the normal control group (P < 0.05). The spleen of the mice in VEGF long-term group was larger than that of the control group, the spleen index also increased (P < 0.05), and remarkable extramedullary hematopoietic signs were found in the HE stained sections. There was no significant change in the total ratio of lymphocytes in the peripheral blood after injection, but the percentage of CD3(+) cells and the CD3(+)/B220(+) ratio in the long-term group deceased; the percentages of Treg and Gr-1(+)CD11b(+) MDSC in the experimental groups increased (P < 0.05), which more significantly increased in the long-term group than that in the short-term group (P < 0.05). It is concluded that the exogenous VEGF promotes hematopoietic stem cell mobilization, and at same time up-regulates the many kinds of suppressive immune cell levels which leads to changes of immuno-function.

Serum deprivation elevates the levels of microvesicles with different size distributions and selectively enriched proteins in human myeloma cells in vitro.

AIM: To investigate the effects of serum deprivation (SD) on microvesicles (MVs) secreted from human myeloma cells and the implications for disease progression. METHODS: RPMI 8226, U266, and KM3 human myeloma cells were incubated in medium containing 10% (non-SD) or 1% fetal bovine serum (SD) and MVs were isolated. The levels and size distribution of MVs were analyzed with flow cytometry. The protein profiles of MVs were studied using 2D SDS-PAGE, MALDI-TOF-MS, and Western blotting. NF-κB activation was analyzed using EMSA. Angiogenesis was examined in Eahy926 endothelial cells. RESULTS: Exposure of RPMI 8226 cells to SD for 24 h did not alter the number of apoptotic cells. However, SD increased the number of MVs from RPMI 8226, U266, and KM3 cells to 2.5-, 4.3-, and 3.8-fold, respectively. The size distribution of SD MVs was also significantly different from that of non-SD MVs. Three proteins ZNF224, SARM, and COBL in SD MVs were found to be up-regulated, which were involved in cell cycle regulation, signal transduction and metabolism, respectively. Co-culture of SD MVs and RPMI 8226 cells increased NF-κB activation in the target RPMI 8226 cells. Furthermore, SD MVs from RPMI 8226 cells significantly increased the microtubule formation capacity of Eahy926 endothelial cells compared with non-SD MVs. CONCLUSION: SD elevates the levels of microvesicles with different size distribution and selectively enriched proteins in human myeloma cells in vitro. The selectively enriched proteins, especially ZNF224, may play key roles in regulation of myeloma cells, allowing better adaptation to SD.

Microvesicles secreted from human multiple myeloma cells promote angiogenesis.

AIM: To investigate whether human multiple myeloma (MM) cells secrete microvesicles (MVs) and whether the MVs secreted from MM cells (MM-MVs) promote angiogenesis. METHODS: RPMI8226 human MM cells and EA.hy926 human umbilical vein cells were used. MVs isolated from RPMI8226 cells were characterized under laser confocal microscopy, electron microscopy and with flow cytometry. The fusion of MM-MVs and EA.hy926 cells was studied under confocal microscopy, and the transfer of CD138 to EA.hy926 cells was demonstrated with flow cytometry. The proliferation, invasion and tube formation of EA.hy926 cells in vitro were evaluated using MTT, transwell migration and tube formation assays, respectively. The vasculization of EA.hy926 cells in vivo was studied using Matrigel plug assay. The expression of IL-6 and VEGF was analyzed with PCR and ELISA. RESULTS: MM-MVs from the RPMI 8226 cells had the characteristic cup-shape with diameter of 100-1000 nm. Most of the MM-MVs expressed phosphatidylserine and the myeloma cell marker CD138, confirming that they were derived from myeloma cells. After added to EA.hy926 cells, the MM-MVs transferred CD138 to the endothelial cells and significantly stimulated the endothelial cells to proliferate, invade, secrete IL-6 and VEGF, two key angiogenic factors of myeloma, and form tubes in vitro and in vivo. CONCLUSION: Our results confirm the presence of MVs in MM cells and support the idea that MM-MVs are newfound mediators for myeloma angiogenesis and may serve as a therapeutic target to treat MM.

Amplification of functional myeloid-derived suppressor cells during stem cell mobilization induced by granulocyte colony-stimulation-factor.

The effects of granulocyte colony-stimulation-factor (G-CSF) on stem cell mobilization and its impact on the amplification of myeloid-derived suppressor cells (MDSCs) of donor mice were examined. A mouse model of stem cell mobilization was established by consecutive subcutaneous injection of 100 µg/kg G-CSF for 5 days. The blood from the donor mice was routinely examined during mobilization. Stem cells and MDSCs were analyzed by flow cytometry. The immunosuppressive molecules derived from MDSCs in serum and spleen, including hydrogen dioxide (H2O2) and nitric oxide (NO), and the activity of nitric oxide synthase (NOS) were determined during the mobilization. Apoptosis of T lymphocytes was assessed by using Annexin-V/PI. During stem cell mobilization, the number of lymphocytes and white blood cells in the peripheral blood was increased, and peaked on the 4th day. The number of stem cells in G-CSF-treated mice was significantly greater than that in controls (P<0.01). The expansions of MSDCs were also observed after G-CSF mobilization, with a more notable rate of growth in the peripheral blood than in the spleen. The activity of NOS and the production of NO were increased in the donor mice, and the serum H2O2 levels were approximately 4-fold greater than the controls. Consequently, apoptosis of T lymphocytes was increased and showed a positive correlation with the elevated percentage of MDSCs. It was concluded that G-CSF could provide sufficient peripheral blood stem cells for transplantation. Exogenous administration of G-CSF caused the accumulation of MDSCs in the peripheral blood and the spleen, which could lead to apoptosis of T lymphocytes and may offer a new strategy for the prevention and treatment of graft versus host disease.

[Research progress of mesenchymal stem cell-derived microvesicle].

Mesenchymal stem cell-derived microvesicle (MSC-MV) is a membrane secretory system which includes microparticle and exosome, and MSC-MV is released by MSC in resting or activated state. MSC-MV selectively package the biological active substances such as lipids, proteins, mRNA and miRNA but not loads them randomly. It has definitive effect of reducing tissue injury, promoting morphological and functional recovery of the injured tissue, and this effect is probably mediated by miRNA. What is more, the MSC-MV may also possess the biological function of immunological regulation, modulation of cell growth and differentiation. The generation, constitution, and function of MSC-MV are reviewed in this article.

[Metabolic syndrome and microalbuminuria accompanied with hyperglycemia in community subjects].

OBJECTIVE: To examine the prevalence of metabolic syndrome (MS) and microalbuminuria (MAU) in subjects with different levels of glucose tolerance and probe the risk factors for the development of MAU. METHODS: A total of 951 subjects were divided into 3 groups according to the results of oral glucose tolerance test (OGTT). Among them, there were 674 subjects with normal glucose tolerance (NGT), 195 with impaired glucose regulation (IGR) and 82 newly-diagnosed cases of type 2 diabetes mellitus (T2DM). MAU was diagnosed if urine albumin-to-creatinine ratio (UACR) was 30 - 300 mg/g. RESULTS: (1) Compared to the NGT subjects, both the IGR and newly-diagnosed T2DM subjects had the significantly higher levels of age, body mass index (BMI), waist circumference, fasting plasma glucose, 2-hour post OGTT glucose (2 hPG), glycated hemoglobin, glycated albumin, fasting insulin, homeostasis model of assessment-insulin resistance (HOMA-IR), triglyceride, systolic blood pressure (SBP) and UACR but a lower level of high density lipoprotein-cholesterol (HDL-C); (2) The incident rate of MS and MAU in the IGR and newly-diagnosed T2DM subjects was 54.4% (106/195), 12.3% (24/195) and 61.0% (50/82), 12.2% (10/82) respectively versus 9.1% (61/674) and 4.9% (33/674) in the NGT subjects. The incident rate of abdominal obesity, hypertension, hypertriglyceridemia, low HDL-C, MS and MAU in the IGR and newly-diagnosed T2DM subjects was significantly higher than that of the NGT subjects (all P < 0.05); (3) Multiple regression analysis showed that BMI, SBP and 2 hPG were independently associated with UACR. Logistic regression analysis indicated that BMI, SBP and 2 hPG were independent risk factors of MAU. CONCLUSIONS: The prevalence of MS and MAU is significantly higher in the IGR and newly-diagnosed T2DM subjects. BMI, SBP and 2 hPG are independent risk factors of MAU.