Cordycepin Augments the Chemosensitivity of Human Glioma Cells to Temozolomide by Activating AMPK and Inhibiting the AKT Signaling Pathway

IntroductionThe long non-coding RNA HULC has been shown to be involved in the development of several human cancers. The present study was undertaken to investigate the regulatory role of lncRNA-HULC in growth and metastasis of human glioma.Material and methodsThe gene expression of lncRNA-HULC was estimated from the clinical glioma tissues and cell lines using RT-PCR. The proliferation of transfected cancer cells was determined with the help of cell counting kit-8 (CCK8). DAPI staining and dual annexin V-FITC/PI staining procedures were used for inferring the apoptosis of transfected cancer cells. Scratch-heal and transwell chamber assays were employed for the determination of migration and invasion of transfected cells. The expression of proteins of interest was studied by western blotting technique.ResultsThe results showed that lncRNA-HULC exhibits significantly (p < 0.05) higher expression in glioma tissues and cancer cells. The knockdown of lncRNA-HULC led to a marked decline in the proliferation of glioma cells through apoptotic induction which was accompanied by upregulation of Bax and downregulation of Bcl-2. Moreover, knockdown of lncRNA-HULC significantly (p < 0.05) suppressed the migration and invasion of cancer cells in vitro. The western blot analysis showed that lncRNA-HULC exerted its effects via modulation of the PI3K/AKT signaling pathway.ConclusionsThe study revealed the possibility of targeting the PI3K/AKT signaling pathway in glioma through transcriptional knockdown of lncRNA-HULC, which might be utilized for therapeutic purposes against human glioma.

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LRIG1 Modulated Radioresistance of Glioma via Regulating CTLA-4/AKT Signaling Pathway

10.21203/rs.3.rs-131093/v1 ◽

2020 ◽

Author(s):

Shiqi Cheng ◽

Xiangqun Huang ◽

Raorao Yuan ◽

Yan Zhang

Keyword(s):

Correlation Analysis ◽

Western Blot ◽

Signaling Pathway ◽

Glioma Cells ◽

Annexin V ◽

Akt Signaling ◽

Potential Mechanism ◽

Human Glioma ◽

Akt Signaling Pathway ◽

Glioma Tissue

Abstract Background: Radioresistance has a great impact on prognosis of glioma patients. However, the potential mechanism underlying the radioresistance of glioma cells remains largely unknown. Methods: LRIG1 overexpression model was firstly established by using Flag-LRIG1 plasmid. The expression of LRIG1, CTLA-4 proteins were detected by western blot and IHC in cells and human tissue. Real-time PCR was used for deterring mRNA expression. Cell viability and apoptosis were detected using CCK-8 and Annexin-V/propidium iodide (PI), respectively. Co-Immunoprecipitation was used for detecting the combination of LRIG1 and CTLA-4 proteins. Results: LRIG1 was significantly down-regulated in radioresistant glioma cells. Overexpressed LRIG1 could promote the radiosensitivity of glioma cells, meanwhile, inhibit the expression of p-AKT and CTLA-4 protein in radioresistant glioma cells. Furthermore, LRIG1 combined with CTLA-4 and promoted CTLA-4 degradation. In human glioma tissue, LRIG1 was down-regulated, while CTLA-4 was highly expressed in glioma tissue. Finally, correlation analysis showed that the expression of LRIG1 was negatively correlated with expression of CTLA-4 and radioresistance of glioma patients. Conclusion: Our findings demonstrated that LRIG1 facilitates radioresistance glioma cells by regulating CTLA4 /AKT signaling pathway.

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