CBMT-18. THE ROLE OF BIOMARKER CANDIDATE GELSOLIN AND ITS MICRORNAS IN GLIOBLASTOMA

Abstract BACKGROUND Among potential glioblastoma (GBM) blood biomarkers that we identified recently, we focused on gelsolin (GSN), a key regulator of actin filament disassembly. GSN was significantly lower in the blood of patients with GBM than in that of healthy controls. In this study, we analyzed the function of GSN and identified microRNAs (miRs) involved in GSN expression in GBM. METHODS QRT-PCR and western blot were introduced to evaluate the expression level of GSN in normal brain and GBM tissue. The localization of GSN was examined by immunohistochemistry and immunocytochemistry using human samples. The association between the expression level of GSN and progression free survival (PFS) /overall survival (OS) in GBM was assessed by Kaplan-Meier analysis. The function of GSN and its signal transduction in glioma cell lines were analyzed using small interfering RNA (siRNA) knockdown of GSN. Additionally, miRs controlling GSN expression were retrieved from databases of miRs, and miRs related to GSN expression were identified in GBM tissues. RESULTS The expression level of GSN was significantly lower in GBM tissues compared to normal brains. Normal astrocytes mainly expressed GSN. High expressor of GSN showed longer PFS and OS than low expressor. Proliferation and invasion in glioma cell lines were significantly promoted by siRNA for GSN accompanied with the activation of glycogen synthase kinase 3β. In GBM tissues, the expression levels of GSN and miR-654-5p, 450b-5p showed an inverse correlation. The inhibitor for miR-654-5p and miR-450b-5p accelerated GSN expression resulting reduction of proliferation. CONCLUSION GSN plays a role as suppressor of proliferation and invasion in GBM. miR-654-5p and miR-450b-5p which control GSN expression can be targets against GBM.

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Long Intergenic Noncoding RNA 00152 Promotes Glioma Cell Proliferation and Invasion by Interacting with MiR-16

Cellular Physiology and Biochemistry ◽

10.1159/000488836 ◽

2018 ◽

Vol 46 (3) ◽

pp. 1055-1064 ◽

Cited By ~ 12

Author(s):

Xin Chen ◽

Deheng Li ◽

Yang Gao ◽

Wei Tang ◽

Lao IW ◽

...

Keyword(s):

Cell Proliferation ◽

Cell Lines ◽

Glioma Cell ◽

Migration And Invasion ◽

Human Glioma ◽

Human Glioma Cell ◽

Glioma Cell Lines ◽

Glioma Cell Proliferation ◽

Proliferation And Invasion

Background/Aims: Long noncoding RNAs (lncRNAs) are a novel class of protein-noncoding transcripts that are aberrantly expressed in multiple diseases including cancers. LINC00152 has been identified as an oncogene involved in many kinds of cancer; however, its expression pattern and function in human glioma remain unclear. Methods: Quantitative real-time polymerase chain reaction was carried out to measure LINC00152 expression in human glioma cell lines and tissues. CCK-8 and EdU assays were performed to assess cell proliferation, and scratch assays and Transwell assays were used to assess cell migration and invasion, respectively. Luciferase reporter assays were carried out to determine the interaction between miR-16 and LINC00152. In vivo experiments were conducted to assess tumor formation. Results: LINC00152 was found to be significantly upregulated in human glioma cell lines and clinical samples. Knockdown of LINC00152 suppressed glioma cell proliferation, migration, and invasion in vitro. In vivo assays in nude mice confirmed that LINC00152 knockdown inhibits tumor growth. Furthermore, mechanistic investigation showed that LINC00152 binds to miR-16 in a sequence-specific manner and suppresses its expression. miR-16 inhibition strongly attenuated LINC00152 knockdown–mediated suppressive effects on proliferation, migration, and invasion. Moreover, LINC00152 induced BMI1 expression by sponging miR-16; this effect further promoted glioma cell proliferation and invasion. Conclusion: We regard LINC00152 as an oncogenic lncRNA promoting glioma cell proliferation and invasion and as a potential target for human glioma treatment.

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Migration, proliferation and invasion behaviour of re-irradiated glioma spheroids using two different human glioma cell lines

Clinical Neurology and Neurosurgery ◽

10.1016/s0303-8467(97)81865-3 ◽

1997 ◽

Vol 99 ◽

pp. S132

Author(s):

H. Arnold ◽

J. Gliemroth ◽

T. Feyerabend ◽

U. Knopp ◽

A.J.A. Terzis

Keyword(s):

Cell Lines ◽

Glioma Cell ◽

Human Glioma ◽

Human Glioma Cell ◽

Glioma Cell Lines ◽

Proliferation And Invasion

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Overexpression of DcR3 and Its Significance on Tumor Cell Differentiation and Proliferation in Glioma

The Scientific World JOURNAL ◽

10.1155/2014/605236 ◽

2014 ◽

Vol 2014 ◽

pp. 1-7 ◽

Cited By ~ 2

Author(s):

Suning Huang ◽

Gang Chen ◽

Yiwu Dang ◽

Long-Hua Chen

Keyword(s):

Tumor Cell ◽

Cell Lines ◽

Glioma Cell ◽

Glioma Cells ◽

Normal Brain ◽

Ki 67 ◽

Glioma Cell Lines ◽

Positive Correlations ◽

Differentiation And Proliferation ◽

Tumor Cell Differentiation

Background.Overexpression of decoy receptor 3 (DcR3) have been reported in various classes of malignancies. However, its expression and clinicopathological contribution in gliomas has not been fully elucidated.Objective.To explore the expression and clinical significance of DcR3 protein in relation to tumor cell differentiation and proliferation in glioma cell lines and tissues.Methods.One hundred and twenty-five samples of glioma patients and 18 cases of normal brain tissues were recruited. The expression of DcR3 protein was detected using immunohistochemistry. Tumor differentiation was assessed by histologic characters and the status of glial fibrillary acidic protein (GFAP). Tumor cell labeling indexes (LIs) of Ki-67 and PCNA were also obtained. The relationship between the DcR3 level and clinicopathological features was investigated, including tumor differentiation, LIs, and survival. Meanwhile, the expression of DcR3 protein was also measured in the supernatants of 8 glioma cell lines and glioma cells freshly prepared from 8 human glioblastoma specimens by using western blot.Results.The level of DcR3 protein in gliomas was significantly higher than that in normal brain tissues (P<0.01). DcR3 expression showed positive correlations with tumor pathological grade(r=0.621,P<0.01)and negative with GFAP expression (r=-0.489,P<0.01). Furthermore, there were positive correlations between DcR3 expression and Ki-67, PCNA LIs (r=0.529,P<0.01;r=0.556,P<0.01). The survival in the DcR3 negative group was 50 ± 1.79 months, longer than that of the DcR3 positive group (48.36 ± 2.90), however, without significance(P=0.149). Different levels of DcR3 could also be detected in the culturing supernatants of all the 8 glioma cell lines and glioma cells freshly obtained from 8 human glioblastoma specimens.Conclusions.The overexpression of DcR3 might play a crucial role in the tumorigenesis, differentiation, and proliferation of glioma.

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Long Noncoding RNA H19 Promotes Proliferation and Invasion in Human Glioma Cells by Downregulating miR-152

Oncology Research Featuring Preclinical and Clinical Cancer Therapeutics ◽

10.3727/096504018x15178768577951 ◽

2018 ◽

Vol 26 (9) ◽

pp. 1419-1428 ◽

Cited By ~ 18

Author(s):

Lei Chen ◽

Yuhai Wang ◽

Jianqing He ◽

Chunlei Zhang ◽

Junhui Chen ◽

...

Keyword(s):

Cell Proliferation ◽

Cell Lines ◽

Glioma Cell ◽

Glioma Cells ◽

Human Glioma ◽

Human Glioma Cell ◽

Glioma Cell Lines ◽

Glioma Cell Proliferation ◽

Proliferation And Invasion

miR-152 and lncRNA H19 have been frequently implicated in various cellular processes including cell proliferation, invasion, angiogenesis, and apoptosis. However, the interaction between miR-152 and H19 in glioma has never been reported. RT-qPCR was used to examine the expression of miR-152 and H19 in human glioma cell lines and normal human astrocytes (NHAs). The interaction between miR-152 and lncRNA H19 was assessed by dual-luciferase reporter assay. MTT assay and Transwell invasion assay were used to determine the proliferation and invasion of U251 and U87 cells. A xenograft tumor experiment was performed to confirm the role of H19 in vivo. The results showed that H19 expression was upregulated and miR-152 expression was downregulated in human glioma cell lines. H19 downregulation or miR-152 upregulation suppressed glioma cell proliferation and invasion in vitro. Moreover, H19 and miR-152 directly regulated each other. Furthermore, decreased miR-152 expression alleviated si-H19-induced inhibitory effects on proliferation and invasion in glioma cells. As expected, H19 silencing hindered glioma growth in vivo. Taken together, H19 promoted glioma cell proliferation and invasion by negatively regulating miR-152 expression, providing evidence for the potential application of H19 as a biomarker and therapy target for glioma.

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Circular RNA hsa-circ-0012129 Promotes Cell Proliferation and Invasion in 30 Cases of Human Glioma and Human Glioma Cell Lines U373, A172, and SHG44, by Targeting MicroRNA-661 (miR-661)

Medical Science Monitor ◽

10.12659/msm.909229 ◽

2018 ◽

Vol 24 ◽

pp. 2497-2507 ◽

Cited By ~ 16

Author(s):

Gang Xie

Keyword(s):

Cell Proliferation ◽

Cell Lines ◽

Glioma Cell ◽

Circular Rna ◽

Human Glioma ◽

Human Glioma Cell ◽

Glioma Cell Lines ◽

Proliferation And Invasion

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Proteolipid Protein 2 Overexpression Indicates Aggressive Tumor Behavior and Adverse Prognosis in Human Gliomas

International Journal of Molecular Sciences ◽

10.3390/ijms19113353 ◽

2018 ◽

Vol 19 (11) ◽

pp. 3353 ◽

Cited By ~ 8

Author(s):

Yi-Hsuan Chen ◽

Dueng-Yuan Hueng ◽

Wen-Chiuan Tsai

Keyword(s):

Cell Proliferation ◽

Cell Lines ◽

Glioma Cell ◽

Proteolipid Protein ◽

Migration And Invasion ◽

Normal Brain ◽

Human Gliomas ◽

Advanced Tumor ◽

Glioma Cell Lines ◽

Aggressive Tumor

Proteolipid protein 2 (PLP2), a membrane protein of the endoplasmic reticulum, is related to tumor proliferation and metastasis in some human cancers, but not in gliomas. First, we performed western-blot analysis, real-time quantitative PCR and immunohistochemical stains to detect PLP2 expression in 4 glioma cell lines and human glioma tissues. In addition, we used small interfering RNA (SiPLP2) and short hairpin RNA (shPLP2) to knockdown PLP2 expression in GBM8401 and LN229 glioma cell lines. After then, the alteration of PLP2 suppressed glioma cells behavior were examined by cell proliferation, wound healing, cell invasion, and colonies formation assays. Finally, the possible mechanism of PLP2 was analyzed by detecting the expression of the proteins related to cell-cycle checkpoints, cell-proliferative signaling factors, and cell-matrix interaction. Compared with normal brain cell lysates and mRNA, all glioma cell lines displayed PLP2 protein and mRNA overexpression. Besides, higher PLP2 IHC staining significantly correlated with more advanced tumor grades and poorer prognosis in human gliomas. Both siPLP2 transfected gliomas showed a clear inhibition of glioma cell proliferation, migration, and invasion as well as down-regulating p-p38, p-ERK, MMP-2, and MMP-9 expression. In conclusion, we successfully demonstrated that PLP2 overexpression played an oncogenic role in glioma development and aggressive tumor behavior.

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ARID4B is a good biomarker to predict tumour behaviour and decide WHO grades in gliomas and meningiomas

Journal of Clinical Pathology ◽

10.1136/jclinpath-2016-203804 ◽

2016 ◽

Vol 70 (2) ◽

pp. 162-167 ◽

Cited By ~ 7

Author(s):

Wen-Chiuan Tsai ◽

Dueng-Yuan Hueng ◽

Shin Nieh ◽

Hong-Wei Gao

Keyword(s):

Brain Tissue ◽

Cell Lines ◽

Glioma Cell ◽

Statistical Significance ◽

Normal Brain ◽

Who Grade ◽

Tumour Metastasis ◽

Glioma Cell Lines ◽

Immunostaining Score

AimsAlthough ARID4B is known to promote tumour metastasis in breast cancer and inhibit transformation and progression in leukaemia, the possible effect of ARID4B on primary brain tumours (PBTs) is not well characterised. We tested the hypothesis that expression of ARID4B correlates with WHO grade and survival in patients with PBTs.MethodsWestern blot analysis was performed on protein lysates prepared from normal brain tissue and glioma cell lines (U87MG, LN229, GBM8401 and U118MG). Subsequently, immunohistochemical analysis of ARID4B was performed on 2 tissue microarrays, including 12 normal brain tissues, 63 meningiomas with different subtypes, 232 gliomas of various grades and degrees of differentiation, 8 central neurocytomas and 4 chordomas. The ARID4B immunostaining score was calculated by multiplying the intensity score by the percentage of tumour cells expressing ARID4B.ResultsIn vitro, ARID4B protein expression was increased in some glioma cell lines. In addition, the average ARID4B immunostaining score was 38.03, 79.09, 129.76 and 119.32, respectively, in gliomas of WHO grade I, II, III and IV. Higher ARID4B immunostaining score was significantly correlated with more advanced WHO grade of gliomas (p=7.4×10–6) and meningiomas. Finally, higher ARID4B expression tended to the shorter survival rates, but did not reach statistical significance.ConclusionsARID4B overexpression presented in most of PBTs, rather than non-neoplastic brain tissue, and correlated with WHO grades in meningiomas and gliomas. Therefore, ARID4B is a satisfactory biomarker to highlight tumour component and predict tumour behaviour in primary brain neoplasms.

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Knockdown lncRNA CRNDE enhances temozolomide chemosensitivity through autophagy inhibition by activating PI3K/Akt/mTOR pathway in glioblastoma

10.21203/rs.3.rs-544906/v1 ◽

2021 ◽

Author(s):

Zijin Zhao ◽

Miaomiao Liu ◽

Wenyong Long ◽

Jian Yuan ◽

Haoyu Li ◽

...

Keyword(s):

Cell Viability ◽

Cell Lines ◽

Glioma Cell ◽

Mtor Pathway ◽

Normal Brain ◽

Mouse Tumor ◽

Qrt Pcr ◽

Potential Biomarker ◽

Glioma Cell Lines ◽

Related Proteins

Abstract Background Long non-coding RNA (lncRNA) CRNDE is highly expressed in glioma acting as an oncogene in gliomas. However, the regulatory roles of CRNDE in Temozolomide (TMZ) chemoresistance to glioblastoma multiforme (GBM) still are poorly understood. Therefore, we intended to explore the role and possible mechanism of CRNDE in TMZ-induced chemoresistance in GBM. Methods Firstly, we tested the expression level of CRNDE in glioma cell lines and in 30 cases of normal brain tissues and 58 cases of glioma tissue specimens by qRT-PCR. Meanwhile, the correlation between CRNDE level and the clinicopathological characteristics and survival time of patients were analyzed. Then, the CRNDE expression in various glioma cell lines was detected, and CRNDE knockdown cell models were constructed. Subsequently, to explore the effect of CRNDE on chemosensitivity to TMZ, cell viability was detected by the CCK-8 assay and IC50 values, and cell proliferation was detected by cell clone assay and EdU assay, as well as cell survival was detected by apoptosis with flow cytometry under TMZ treatment. Further, the expression of drug-resistance protein ABCG2, autophagic related proteins and PI3K/AKT pathway related proteins were determined by western blot or qRT-PCR in TMZ-treated glioma cells. Finally, the tumor volume and weight were measured and ABCG2 expression was detected by immunohistochemistry assay in mouse tumor xenograft model. Results Our integrated approach demonstrated lncRNA CRNDE was a poor prognosis factor for GBM patient, which was up-regulated in patients who are resistant to TMZ, and closely associated with chemotherapeutic response status to TMZ treatment. Further, functional assays revealed that knockdown of CRNDE could notably reduce glioma cell viability and proliferation, and elevate apoptosis to enhance the chemosensitivity to TMZ in vitro and in vivo. Mechanistically, the depression of CRNDE down-regulated LC3 II/I, Beclin1 and Atg5 expression levels and increased expression of p62 to inhibit autophagy due to the activation of PI3K/Akt/mTOR pathway as well as highly correlated with ABCG2 expression. Conclusions Overall, the study provided a preclinical basis in developing LncRNA CRNDE as a reliable clinical predictor of outcome and prognosis of GBM patient, a potential biomarker for predicting TMZ treatment response and a new therapeutic target for enhancing TMZ sensitivity in GBM by modulating the autophagy through PI3K/Akt/mTOR pathway.

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Serum lncRNA-ANRIL and SOX9 expression levels in glioma patients and their relationship with poor prognosis

World Journal of Surgical Oncology ◽

10.1186/s12957-021-02392-2 ◽

2021 ◽

Vol 19 (1) ◽

Author(s):

Youlu Sun ◽

Yuesong Jing ◽

Yuxin Zhang

Keyword(s):

Cell Lines ◽

Glioma Cell ◽

Tumor Grade ◽

Test Cell ◽

Control Group ◽

Normal Brain ◽

Tumor Diameter ◽

Western Blot Assay ◽

Bax Protein ◽

Glioma Cell Lines

Abstract Background lncRNA-CDKN2B antisense RNA 1 (ANRIL) and SRY-box transcription factor 9 (SOX9) has abnormal expression in many tumors including glioma, but the underlying molecular mechanism is unclear. This study set out to investigate the serum lncRNA-ANRIL and SOX9 levels in glioma patients and their effects on prognosis. Methods We enrolled 142 glioma patients admitted to our hospital from May 2014 to May 2016 into the research group (RG) and 120 healthy subjects receiving concurrent physical examinations into the control group (CG). Fasting peripheral blood (4 mL each) was sampled from subjects from the two groups. Using the quantitative real-time polymerase chain reaction (qRT-PCR), lncRNA-ANRIL and SOX9 were measured to explore their values in the early diagnosis of glioma. Patients from RG were followed up for 3 years to analyze the influence of lncRNA-ANRIL and SOX9 on patient prognosis. We purchased glioma cell lines U251 and U87 and grouped them according to the transfection of different plasmids. We conducted CCK8 assay to test cell proliferation, Transwell assay to test cell invasion, the flow cytometry to test cell apoptosis, and Western Blot assay to measure bcl-2 and bax protein levels. Results ANRIL and SOX9 were evidently higher in RG than in CG (P<0.01). The receiver operating characteristic (ROC) curve revealed that the diagnostic sensitivity of ANRIL combined with SOX9 for glioma was 81.62%, and the specificity was 90.83% (P<0.01). ANRIL and SOX9 were closely related to tumor grade, tumor diameter, distant metastasis, and family history of glioma (P<0.01). In total, 135 patients were successfully followed up (95.07%). Patients with high levels of ANRIL and SOX9 had a markedly poorer prognosis than those with low levels (P<0.05). ANRIL and SOX9 were markedly higher in glioma cell lines (U251 and U87) than in normal brain cells (P<0.01). The proliferation and invasion of U251 cells were notably reduced after the transfection of ANRIL and SOX9 inhibitory sequences (P<0.01), but the apoptosis was notably increased (P<0.01). Bcl-2 expression was markedly increased in lncRNA-ANRIL-inhibitor and SOX9-inhibitor (P<0.01), while bax expression was markedly reduced in lncRNA-ANRIL-inhibitor and SOX9-inhibitor (P<0.01). Conclusion lncRNA-ANRIL and SOX9 levels were higher in glioma patients than in healthy people. High-lncRNA-ANRIL and SOX9 levels were strongly associated with unfavorable prognosis of patients. The testing of biological behaviors revealed that lncRNA-ANRIL and SOX9 worked as tumor-promoting genes in glioma.

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