Up-regulation of circular RNA hsa_circ_01844 induces apoptosis and suppresses proliferation and migration of glioblastoma cells.

BACKGROUND: Previous studies have demonstrated that various circular RNAs are involved in the malignant proliferation of cancers, such as liver cancer, lung cancer, breast cancer, and others. The potential role of circular RNAs in glioblastoma, however, is still uncertain. In this study, we aimed to study the potential role of hsa_circ_01844 in glioblastoma. METHODS: Using reverse transcription-polymerase chain reaction (RT-PCR) method, hsa_circ_01844 expression was measured in five glioblastoma samples and five normal brain samples. To evaluate the potential function of hsa_circ_01844 in glioblastoma, hsa_circ_01844 was overexpressed in glioblastoma cell lines (U251 and U87 cells). Using these two cell lines, in vitro experiments including the flow cytometry assay, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay, Transwell assay, and cell apoptosis assay were performed to investigate the role of hsa_circ_01844 in glioblastoma. Student t test and one-way analysis of variance were used for statistical analysis. RESULTS: The expression of circular RNA hsa_circ_01844 was lower in glioblastoma tissues when compared with the normal brain tissues by RT-PCR method (0.034 ±â€Š0.036 vs. 1.630 ±â€Š0.891, P < 0.001). Using two glioblastoma cell lines, we found that overexpression of hsa_circ_01844 in glioblastoma cells suppressed their proliferation, colony formation, migration, and increased the apoptotic rate compared with empty vector group and blank control group (all P < 0.05). CONCLUSION: Hsa_circ_01844 shows decreased expression in glioblastoma and its overexpression induces apoptosis and inhibits proliferation, migration, and invasion of glioblastoma cells.

Overexpression of G-protein-coupled receptors 65 in glioblastoma predicts poor patient prognosis.

OBJECTIVE: G-protein-coupled receptors 65 (GPR65), identified as an acid-sensing receptor, is overexpressed in several malignancies and promote tumor development. Our aim was to investigate the expression and prognostic value of GPR65 in glioblastoma. MATERIALS AND METHODS: We determined the expression of GPR65 protein using immunohistochemistry in tissue microarrays containing 11 Grade I, 107 Grade II, 47 Grade III, and 102 Grade IV gliomas and 16 normal brains. Then we evaluated its association with pathological grades, prognosis, and recurrence. The Cancer Genome Atlas (TCGA) group (N=528) was further employed to examine transcriptional level of GPR65 in glioblastoma and the correlation between GPR65 expression and clinical outcome. RESULTS: In our cohort, GPR65 expression was positively related to glioma pathological grade (p<0.01) and elevated in glioblastoma (p<0.01). High expression of GPR65 was associated with significantly short overall survival (OS) (p=0.013) and progression-free survival (PFS) (p=0.029), and could be identified as an independent risk factor for OS of glioblastoma patients (Hazard Ratio [HR]=1.596, p=0.037). As an aiding evidence, increased GPR65 mRNA expression was also found in TCGA glioblastoma group (p<0.001) and its high level predicted a poor clinical outcome (OS, p=0.003; PFS, p=0.001). CONCLUSION: Our findings suggest that GPR65 is overexpressed in glioblastoma and its high expression predicts unfavorable clinical outcome for patients. Targeting GPR65 may serve as a potential therapy for treating glioblastoma.

[Functional improvement of patients with progressive muscular dystrophy by bone marrow and umbilical cord blood mesenchymal stem cell transplantations].

OBJECTIVE: To investigate the feasibility of employing double transplantations of autologous bone marrow mesenchymal stem cells (BMSC) and umbilical cord mesenchymal stem cells (UMSC) in the treatment of progressive muscular dystrophy (PMD). METHODS: A total of 82 cases were treated by the double transplantations of BMSC and CB-MSC. They were diagnosed by clinical manifestations, CK, LDH, genetic analysis, electromyography, MRI and pathologic examination of biopsied muscle specimens from July 2007 to July 2008. Control group was self-made at before and after treatment and cases were followed up for 3 - 12 months. treatment method: Eighty-two patients underwent the double transplantations of bone mesenchymal stem cell (BMSC) and human umbilical cord blood MSC (CB-MSC). (1) BMSC: 80 - 150 ml bone marrow sample was collected through a puncture at bilateral posterior superior iliac spine. Ficoll density gradient centrifuge was employed to separate individual monocyte for induced differentiation. (2) CB-MSC: 80 - 160 ml umbilical cord blood was harvested and processed likewise as above. (3) Stem cell transplantation: Both BMSC and CB-MSC were collected and prepared into 1 x 10(8)/ml and 1 x 10(7)/ml cell suspension respectively. They were transplanted in divided does into the extremity muscle and vein. The clinical and laboratory parameters were monitored at 3, 6, 9 and 12 months. RESULTS: It was found that 31 cases (37.8%) obtained a remarkable efficacy, 37 cases (45.1%) were effective and 14 cases (17.1%) had no change. Total effective rate was 82.9%. Seventy patients (85.4%) felt limbs warmly, appetite improved, gained weight, had better appetite and action were nimble. Activity of daily living scale (ADL) in 72 patients (87.8%) increased as compared with pre-treatment (P < 0.01). LDH decreased at post-treatment [(475 +/- 223) u/L vs (410 +/- 216) u/L, P < 0.05, t = 6.650]. Creatine kinase [(2952 +/- 2259) u/L vs (2841 +/- 2092) u/L, P = 0.223, t = 1.094] and creatine [(26 +/- 12) micromol/L vs (25 +/- 11) micromol/L, P = 0.306, t = 1.029] decreased slightly. Adherence to therapy among Children and no adverse reaction was reported during the course of treatment. CONCLUSION: The double transplantation of BMSC and CB-MSC is convenient, safe and effective in the treatment of progressive muscular dystrophy and can be considered as a new therapy of PMD. MSC represents a possible tool of cellular therapeutics for PMD.