LncRNA LINC01410 Induced by MYC Accelerates Glioma Progression via Sponging miR-506-3p and Modulating NOTCH2 Expression to Motivate Notch Signaling Pathway.

Glioma is a common invasive cancer with unfavorable prognosis in patients. Long non-coding RNAs have been reported to participate in modulating diverse cellular processes. Here, we focused on exploring the role of long intergenic non-protein coding RNA 1410 (LINC01410) in glioma and its underlying mechanism. The expression levels and protein levels of genes were analyzed by quantitative real-time PCR (RT-qPCR) analysis and western blot. Loss-of-function assays were performed to assess the function of LINC01410 in glioma cells. The interactions among MYC, LINC01410, microRNA-506-3p (miR-506-3p) and notch receptor 2 (NOTCH2) were validated through Chromatin immunoprecipitation (ChIP), RNA Binding Protein immunoprecipitation (RIP), RNA pull-down and luciferase reporter assays. Our data supported that LINC01410 was up-regulated in glioma cells. Bioinformatics predictions and the integrated experiments identified that MYC activated LINC01410 transcription and LINC01410 promoted the levels of NOTCH2 through sponging miR-506-3p and further motivated Notch signaling pathway. Rescue assays validated that LINC01410 exerted its influential functions on glioma cell proliferation and apoptosis via enhancing NOTCH2 expression. Our studies identified that LINC01410 accelerates the progression of glioma through acting as a ceRNA for miR-506-3p and elevating NOTCH2 expression to further activate the Notch signaling pathway, which indicated that LINC01410 might act as a novel regulator of glioma progression.

CREB1-induced miR-1204 promoted malignant phenotype of glioblastoma through targeting NR3C2.

BACKGROUND: Glioblastoma (GBM) is a subclass of brain malignancy with unsatisfactory prognosis. MicroRNAs (miRNAs) are a group of non-coding RNAs (ncRNAs) that exert key function on tumorigenesis and tumor development. PURPOSES: The purpose of this work was to unravel the biological behavior and mechanism of miR-1204 in GBM. METHODS: Expressions of miR-1204, NR3C2 and CREB1 were detected by RT-qPCR and western blot. Proliferation and apoptosis of GBM cells were detected by CCK-8, colony formation, caspase-3 activity and TUNEL assays. Molecular interplays were examined by ChIP, RIP, and luciferase reporter assays. RESULTS: MiR-1204 level was elevated in GBM cell lines. Functionally, miR-1204 aggravated cell proliferation whereas suppressed cell apoptosis in GBM cells. Mechanistically, cAMP Responsive Element Binding Protein 1 (CREB1) bound to the promoter of miR-1204 and activated the transcription of miR-1204. Furthermore, miR-1204 targeted and inhibited Nuclear receptor subfamily 3 group C member 2 (NR3C2), a tumor suppressor gene in GBM cells. Rescue assays indicated that NR3C2 participated in the regulation of miR-1204 on the malignant phenotype of GBM cells. CONCLUSIONS: We observed for the first time that CREB1-induced miR-1204 promoted malignant phenotype of GBM through targeting NR3C2, indicating that miR-1204 acted as a novel oncogenic miRNA in GBM.

Long noncoding RNA FOXD2-AS1 promotes glioma malignancy and tumorigenesis via targeting miR-185-5p/CCND2 axis.

Glioma is the most aggressive malignant tumor in the adult central nervous system. Abnormal long noncoding RNA (lncRNA) FOXD2-AS1 expression was associated with tumor development. However, the possible role of FOXD2-AS1 in the progression of glioma is not known. In the present study, we used in vitro and in vivo assays to investigate the effect of abnormal expression of FOXD2-AS1 on glioma progression and to explore the mechanisms. FOXD2-AS1 was upregulated in glioma tissue, cells, and sphere subpopulation. Upregulation of FOXD2-AS1 was correlated with poor prognosis of glioma. Downregulation of FOXD2-AS1 decreased cell proliferation, migration, invasion, stemness, and epithelial-mesenchymal transition (EMT) in glioma cells and inhibited tumor growth in transplanted tumor. We also revealed that FOXD2-AS1 was mainly located in cytoplasm and microRNA (miR)-185-5p both targeted FOXD2-AS1 and CCND2 messenger RNA (mRNA) 3'-untranslated region (3'-UTR). miR-185-5p was downregulated in glioma tissue, cells, and sphere subpopulation. Downregulation of miR-185-5p was closely correlated with poor prognosis of glioma patients. In addition, miR-185-5p mimics decreased cell proliferation, migration, invasion, stemness, and EMT in glioma cells. CCND2 was upregulated in glioma tissue, cells, and sphere subpopulation. Upregulation of CCND2 was closely correlated with poor prognosis of glioma patients. CCND2 knockdown decreased cell proliferation, migration, invasion, and EMT in glioma cells. In glioma tissues, CCND2 expression was negatively associated with miR-185-5p, but positively correlated with FOXD2-AS1. FOXD2-AS1 knockdown and miR-185-5p mimics decreased CCND2 expression. Inhibition of miR-185-5p suppressed FOXD2-AS1 knockdown-induced decrease of CCND2 expression. Overexpression of CCND2 suppressed FOXD2-AS1 knockdown-induced inhibition of glioma malignancy. Taken together, our findings highlight the FOXD2-AS1/miR-185-5p/CCND2 axis in the glioma development.

Short-term therapeutic effects of low-dose cytarabine plus surgical resection on elderly patients with trigeminal nerve tumor and safety observation.

OBJECTIVE: To evaluate the short-term therapeutic effects of low-dose cytarabine plus surgical resection on elderly patients with trigeminal nerve tumor and to observe the safety. METHODS: A total of 120 elderly patients with trigeminal nerve tumor were divided into a treatment group and a control group by random draw (n=60), and both groups were subjected to resection by stereotactic image-guided endoscopic nasal surgery. Afterwards, the control group was administered with high-dose cytarabine while the treatment group was given low-dose cytarabine for 14 days. RESULTS: Both groups completed treatment, but the effective rate of the treatment group (96.7%) was significantly higher than that of the control group (83.3%) (P < 0.05). The pain scores of the two groups were similar at T0, T1 and T2, but the score of the treatment group at T2 was significantly different from those at T0 and T1 (P < 0.05). During treatment, the treatment group was significantly less prone to complications such as headache, vomiting, vision impairment, nausea and local swelling than the control group (P < 0.05). During three months of follow-up, the appetite, sleep and daily living scores were significantly higher than those of the control group (P < 0.05). CONCLUSION: Stereotactic image-guided surgery was able to treat trigeminal nerve tumor well, and the effect was enhanced by low-dose cytarabine that improved postoperative outcomes and quality of life by dramatically decreasing complications.

[Effects of low-frequency electrical stimulation of hippocampus on the expression of GABAA receptor α1 and ß2 subunits in kainate-kindled rats].

OBJECTIVE: To explore the effects of low-frequency electrical stimulation (LFS) of hippocampus on the expression of GABAA receptor α1 and ß2 subunits in the kainite (KA)-kindled rats and explore its underlying rescue mechanism for temporal lobe epilepsy. METHODS: The SD rats were divided into 4 groups (n = 10):control group:no operation; epilepsy group:KA was stereotactically injected into rat hippocampus to establish KA-kindled model; sham-LFS group:only implanting of electrodes into hippocampus without LFS; LFS group:KA-kindled model with LFS. Then hippocampal tissues were collected and their expressions of GABAA receptor α1 and ß2 subunits measured by immunohistochemistry, quantitative-polymerase chain reaction (q-PCR) and Western blot.Nissl staining was performed to detect the number of damaged hippocampal neurons. RESULTS: There were significant decreases of GABAA receptor α1 and ß2 subunits in kindled hippocampus (P < 0.01). LFS resulted in an effective rescue, but failed to reach normal levels. The damaged neurons could not be reversed according to Nissl staining. CONCLUSION: The hippocampal application of LFS may protect against seizures by modulating the expressions of α1 and ß2 GABAA receptor subunits.

[Effects of hippocampal stimulus on α5 subunit of extrasynaptic GABA(A) receptor in kainic acid-induced epileptic rats].

OBJECTIVE: To observe the effects of electrical hippocampal stimulation of α5 subunit of extrasynaptic GABA(A) receptor in kainic acid-induced epileptic rats, explore the optimal therapeutic parameters and elucidate the possible mechanism of electrical stimulation for hippocampal epilepsy. METHODS: A total of 40 healthy male Sprague Dawley (SD) rats were randomly divided into 5 groups of epilepsy, sham operation, low-frequency stimulation (LFS), high-frequency stimulation (HFS) and normal control (n = 8 each). The expression of α5 subunit of extrasynaptic GABA(A) receptor as well as the association with the curative effects of LFS were evaluated by behavioristics, real-time fluorogenic quantitative polymerase chain reaction (PCR) and Western blot. RESULTS: The kainic acid kindling rats had class V attack with a modeling success rate of 67%. After treatment, as compared with sham operation and epilepsy groups, the number of seizures decreased significantly in LFS and HFS treatment groups (P < 0.01). But LFS and HFS treatment groups were compared, the number of seizures has no statistical significance (P > 0.05). When sham operation and epilepsy groups were compared, the number of seizures was not statistically significant (P > 0.05). The paired inter-group comparisons showed that LFS and HFS treatment groups and sham operation and epilepsy groups had significant differences in α5 subunit mRNA and protein (P < 0.01). No statistical significance existed between LFS and HFS treatment groups (P > 0.05) or sham operation and HFS treatment groups (P > 0.05). As compared with normal control group, LFS treatment and HFS groups had significant difference (P < 0.01). CONCLUSION: Electrical hippocampal stimulation can effectively inhibit seizures. Both LFS and HFS are equally effective, but the former offers greater benefits. And an inhibition of extrasynaptic α5 subunit plays an important role in seizure during hippocampal stimulation.

[The effects of low-frequency electric stimulus on hippocampal of Effects of low-frequency electric stimulus on hippocampal of α5 subunit of extra synapse GABAA receptor in kainic acid-induced epilepsy rats].

OBJECTIVE: To explore the effects of low-frequency electric stimulus (LFS) on hippocampal of α5 subunit of extrasynaptic GABAA receptor in kainic acid-induced epilepsy rats and explore the possible mechanism of LFS on hippocampus to treat epilepsy. METHODS: A total of 32 male Sprague-Dawley rats were divided randomly into 4 groups:epileptic, pseudo-surgery, LFS treatment and control. The expression of α5 subunit of extrasynaptic GABAA receptor and the association with the curative effects of LFS were evaluated by behavioristics, real-time fluorogenic quantitative polymerase chain reaction (PCR) and Western blot. RESULT: After treatment, the number of seizures in LFS group were significantly lower than that in the epileptic and pseudo-surgery groups (2.88 ± 0.83 vs 8.50 ± 0.93 and 8.88 ± 0.83) (P < 0.01). The results of real-time PCR demonstrated that the expression of mRNA of α5 subunits in LFS treatment group was obviously higher than that in epileptic and pseudo-surgery groups (0.74 ± 0.20 vs 0.30 ± 0.16 and 0.31 ± 0.16) (P < 0.01), but was lower than 1.10 ± 0.23 in control group. Western blot demonstrated that the protein expression of α5 subunits in LFS (0.75 ± 0.09) and control (0.88 ± 0.09) groups were dramatically higher than that in epileptic (0.22 ± 0.08) and pseudo-surgery (0.26 ± 0.08) groups (P < 0.01). CONCLUSION: Low-frequency electric stimulus on hippocampus is effective in controlling epileptic seizures. And the mechanism may be related with the expression level of α5 subunits of extrasynaptic GABAA receptor.

[Act of Arc siRNA via lentivirus on the memory consolidation of amygdala-kindled rats].

OBJECTIVE: To explore the memory consolidation related role of gene Arc in amygdala-kindled rats. METHODS: A total of 60 male amygdala-kindled rats were divided equally into 3 groups: 0.9% sodium chloride injection, null vector lentivirus injection and silencing lentivirus injection. Quantitative polymerase chain reaction (qPCR) and Western blot were performed to confirm the silencing efficiency of Arc. And the passive avoidance test and Morris' water maze were used to test the memory consolidation ability of rats in three groups. RESULTS: The memory consolidation ability became significantly impaired in the Arc silencing group in both of passive avoidance test and water maze comparing to the other groups. And no significant difference existed in memory consolidation ability between 0.9% sodium chloride and null vector lentivirus injection groups. CONCLUSION: The ability of memory consolidation is impaired in Arc silencing amygdala-kindled rats and Arc may play a significant role in the consolidation of memory in amygdala-kindled rats.

[Expression of α(5) subunit of extrasynaptic GABA(A) receptor in insular-kindled rats].

OBJECTIVE: To explore the expression of α(5) subunit of extrasynaptic GABA(A) receptor in insular electrical kindled rats and analyze its significance. METHODS: A total of 96 male Sprague-Dawley rats were divided randomly into kindled, sham-operated and control groups (n = 32). And each group was further divided into 2 sub-groups at different time points. Kindled group: insular electrical kindled via chronic electrical stimulation; Sham-operated group: electrode implantation without electrical stimulation; CONTROL GROUP: no operation at all. The number of hippocampal neurons was detected by Nissl staining; the mRNA of α(5) subunit of hippocampus by quantitative-polymerase chain reaction (q-PCR); the expression of α(5) subunit of hippocampus by immunohistochemistry. RESULTS: No significant difference existed in the number of hippocampal neurons between epileptic and normal rats (P > 0.05). The mRNA levels of α(5) subunit of hippocampus in the sham-operated and control groups were much higher than that of the kindled group at Day 1 and 7 post-sacrificing (P < 0.01). And the hippocampal expressions of α(5) subunit were much more in the sham-operated and control groups than that of the kindled group at Day 1 and 7 post-sacrificing (P < 0.01). CONCLUSION: The hippocampal expression of α(5) subunit decreases markedly in insular epilepsy. And α(5) subunit may play an important role in the occurrence and development of insular epilepsy.

lncRNA small nucleolar RNA host gene 20 predicts poor prognosis in glioma and promotes cell proliferation by silencing P21.

BACKGROUND: In multiple cancers, long non-coding RNA small nucleolar RNA host gene 20 (lncRNA SNHG20) is generally dysregulated. In the present study, both the biological role and clinicopathological value of lncRNA SNHG20 in glioma are explored. METHODS: Real-time PCR was employed to determine lncRNA SNHG20 expression in glioma patients. The prognostic role of expression of lncRNA SNHG20 was evaluated in a retrospective cohort study. In addition, the association between lncRNA SNHG20 expression and the clinicopathological features of glioma patients, such as tumor recurrence, survival status, follow-up time, WHO grade, resection extent, tumor location, Karnofsky performance scale score, cystic change, tumor size, gender and age, was discussed. By constructing and transfecting siRNAs that targeted lncRNA SNHG20 into the glioma U87 cells, the effects of lncRNA SNHG20 on the proliferation and cell cycle of U87 cells were assessed through cell counting kit-8, colony formation and cell cycle assays, respectively. In addition, Western blot and real-time PCR measured the expression levels of P21 and CCNA1 in U87 cells after being transfected with SNHG20 siRNA. RESULTS: Our results suggested the high expression of lncRNA SNHG20 in human glioma tissues compared with normal brain tissues, which was related to recurrence-free survival and poor overall survival in glioma patients. According to the existing retrospective cohort study, high lncRNA SNHG20 expression was associated with tumor size, extent of resection, WHO grade, follow-up time, survival status and recurrence. Besides, knocking down the expression of lncRNA SNHG20 could inhibit the proliferation and colony formation abilities of glioma U87 cells through cell cycle arrest. Consequently, the expression of CCNA1 was inhibited, and the expression of P21 was up-regulated in U87 cells. CONCLUSION: A high lncRNA SNHG20 expression level predicts the poor prognosis for glioma patients. Moreover, lncRNA SNHG20 can promote glioma proliferation through silencing P21 and thus lncRNA SNHG20 is an independent potential prognostic biomarker for glioma patients.

Cell division cycle associated 7 like predicts unfavorable prognosis and promotes invasion in glioma.

BACKGROUND: Cell division cycle associated 7 like (CDCA7L) belongs to the JPO protein family, which is recently identified as a target gene of c-Myc and is frequently dysregulated in multiple cancers. This study aimed to explore the clinicpathological value and biological role of CDCA7L in glioma. METHODS: CDCA7L expression in glioma patients was determined using the Oncomine database, and the prognostic role of CDCA7L expression was assessed in a retrospective cohort study. Moreover, the relationship of CDCA7L expression with the clinicopathological characteristics in glioma patients, including age, gender, tumor size, cystic change, Karnofsky performance scale (KPS) score, tumor location, extent of resection, WHO grade, adjuvant therapy and tumor recurrence, was analyzed in this study. In addition, the CDCA7L small interfering (si) RNA was constructed and transfected into the glioma U251 cells, so as to examine the role of CDCA7L in glioma patients. Besides, the changes in U251 cell invasion after transfection with CDCA7L siRNA were also monitored through Transwell assay. RESULTS: Our results suggested that CDCA7L expression was up-regulated in different glioma types, including glioblastoma, oligodendroglioma, diffuse astrocytoma and anaplastic astrocytoma. Moreover, the current retrospective cohort study indicated that high CDCA7L expression was associated with tumor size, WHO grade, adjuvant therapy and recurrence, as well as the poor overall survival (OS) and recurrence-free survival (RFS) in glioma patients. Lastly, CDCA7L expression was knocked down using CDCA7L siRNA, which could block the invasion abilities of glioma U251 cells. CONCLUSIONS: CDCA7L is highly expressed in human glioma tissues and a high CDCA7L expression level predicts the dismal prognosis for glioma patients. Moreover, CDCA7L can promote glioma invasion, which can serve as an independent potential prognostic biomarker for glioma patients.

CDCA7L promotes glioma proliferation by targeting CCND1 and predicts an unfavorable prognosis.

Cell division cycle associated 7 like (CDCA7L) belongs to the JPO protein family, recently identified as a target gene of c­Myc and is frequently dysregulated in multiple cancers. However, to the best of our knowledge, no studies to date have been carried out to investigate the functions of CDCA7L in glioma. Thus, in this study, the expression level of CDCA7L and its association with the prognosis in glioma were detected through the TCGA database. The mRNA expression levels of CDCA7L in glioblastoma (GBM) tissues and normal brain tissues were detected by RT­qPCR and western blot analysis. To explore the role of CDCA7L in glioma, CDCA7L siRNA was constructed and transfected into U87 glioma cells. The expression levels of CDCA7L and cyclin D1 (CCND1) in glioma U87 cells following transfection with CDCA7L siRNA were measured by RT­qPCR and western blot analysis. CCK­8, colony formation, EdU and Transwell assays were used to measure the effects of CDCA7L on U87 cell proliferation, and flow cytometry was used to monitor the changes in the cell cycle following transfection with CDCA7L siRNA. Xenograft tumors were examined in vivo for the carcinogenic effects, as well as the mechanisms and prognostic value of CDCA7L in glioma tissues. The results revealed that CDCA7L was highly expressed in human GBM tissues, and a high expression of CDCA7L was associated with a poor prognosis of glioma patients through the TCGA database. We demonstrated that CDCA7L was highly expressed in human GBM tissues and 3 glioma cell lines. The downregulation CDCA7L expression significantly inhibited the proliferation and colony formation ability of U87 cells by blocking cell cycle progression in the G0/G1 phase. In addition, we found that the mRNA and protein levels of CCND1 were markedly decreased following transfection with CDCA7L siRNA compared with NC siRNA in vitro. The downregulation CDCA7L expression reduced the number of invading cells. Consistent with the results of the in vitro assays, the xenograft assay, immunohistochemistry (IHC) assay and western blot analysis demonstrated that, in response to CDCA7L inhibition, tumor growth was inhibited, Ki­67 and CCND1 expression levels were decreased in vivo. On the whole, the results of the current study indicate that CDCA7L is highly expressed in human glioma tissues and that a high CDCA7L expression predicts a poor prognosis of glioma patients. CDCA7L promotes glioma U87 cell growth through CCND1.

GSK1838705A, an IGF-1R inhibitor, inhibits glioma cell proliferation and suppresses tumor growth in vivo.

Glioma is a type of primary malignant tumor of the central nervous system in humans. At present, standard treatment involves surgical resection, followed by radiation therapy and chemotherapy. However, the prognosis is poor and the long­term survival rate remains low. An improved understanding of the molecular basis for glioma tumorigenesis is in urgently required. The pro­survival effect of the insulin­like growth factor (IGF) signaling pathway has been implicated in progression of the glioma disease state. GSK1838705A is a novel, small molecule kinase inhibitor of IGF­IR, which inhibits IGF signal transduction and downstream target activation. Its anti-proliferative activity has been demonstrated in various tumor cell lines. The present study investigated the potential use of GSK1838705A for the treatment of glioma. Human U87MG glioma cells were used to examine the inhibitory activity of GSK1838705A in cell proliferation, migration and apoptosis. The antitumor activity of GSK1838705A was assessed in a xenograft mouse model. GSK1838705A inhibited the growth and induced the apoptosis of the U87MG glioma cells in a dose­dependent manner. The GSK1838705A­treated cells exhibited reduced migratory activity in response to chemoattractants. The present study further demonstrated the antitumor activity of GSK1838705A in vivo. The administration of GSK1838705A significantly inhibited the growth of glioma tumors by inducing the apoptosis of tumor cells. These results suggested that targeting IGF signaling with GSK1838705A may be a promising therapeutic strategy for the treatment of patients with glioma.

Effects of ulinastatin on cerebral oxygen metabolism and CRP levels in patients with severe traumatic brain injury.

The aim of the present study was to investigate the effects of ulinastatin on cerebral oxygen metabolism and C-reactive protein (CRP) levels in patients with severe traumatic brain injury (sTBI). A total of 92 patients with sTBI, admitted to the First Affiliated Hospital of Xinxiang Medical University (Xinxiang, China), were randomly divided into control and observation groups. The control group received conventional therapy plus a placebo (0.9% sodium chloride), while the observation group were administered conventional therapy plus 200,000 units ulinastatin via intravenous injection twice a day for seven days. Arterial and jugular venous blood was collected for blood gas analysis. The jugular venous blood lactate (JVBL), jugular venous bulb oxygen saturation (SjvO2), arteriovenous oxygen content difference (AVDO2) and cerebral extraction of oxygen (CEO2) levels were measured on day 1, 3, 5 and 7, as well as the level of CRP in the peripheral blood. In the control group, the level of JVBL decreased as compared with the level at day 1, however, no statistically significant differences were observed (P>0.05). By contrast, the observation group exhibited a significant reduction in the level of JVBL (P<0.05), which was also significantly lower compared with the control group (P<0.05). Statistically significant differences were observed between the two groups with regard to SjvO2, AVDO2 and CEO2 on day 3, 5 and 7. The CRP levels in the two groups increased and peaked on day 3. However, the CRP level in the observation group significantly decreased on day 5 (35.27±15.18 mg/l) and day 7 (22.65±10.48 mg/l), which was lower compared with the control group (56.19±13.24 mg/l and 47.36±15.73 mg/l, respectively); statistically significant differences were observed (P<0.05). Therefore, ulinastatin effectively improved cerebral oxygen metabolism and reduced the CRP level in patients with sTBI.