PI3K/AKT Signaling Pathway Mediates the Proliferation, Migration and Invasion of Papillary Craniopharyngioma Cells

Abstract Objective:Craniopharyngiomas are rare, histologically benign but clinically challenging neoplasms. Here, we aimed to interrogate the effect and significance of Phosphatidylinositol-3-kinase (PI3K) signaling pathway on papillary craniopharyngioma (PCP) cell growth and survival.Methods: We used Western blotting (WB) experiments to evaluate the expression of the PI3K/protein kinase B (AKT) in Craniopharyngiomas tissues, relative to health tissues. Primary tumor cells were obtained from fresh PCP samples by cell culture and then determined by cell morphology, immunofluorescence staining and expression of specific cell markers. In this study, PCP cell lines, isolated from fresh PCP samples, were treated with different concentrations of LY294002, a PI3K/AKT signaling inhibitor, to evaluate their proliferation, migration and invasion. We determined the cell proliferation using Cell Counting Kit-8 and colony formation. We then used flow cytometry to evaluate cell apoptosis and cell cycle. In addition, cell migration and invasion levels were determined by wound healing and Transwell assays, respectively.Results: Our data demonstrated that the expression of phosphorylated-PI3K/AKT was upregulated in human craniopharyngioma tissues compared to the normal control tissues. Immunofluorescence assays showed the presence of cytokeratin (pan CK) and vimentin protein (VIM) in the PCP primary cells. Furthermore, inhibition of PI3K/AKT signaling blocks the proliferation, migration and invasion of the PCP primary cells.Conclusions:Taken together, our data robustly demonstrates that the PI3K/AKT signaling pathway mediates the proliferation, migration and invasion of the PCP cells.

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LY294002 Inhibit Proliferation, Migration, and Invasion of Craniopharyngiomas via PI3K/Akt Signaling Pathway

10.21203/rs.3.rs-96933/v1 ◽

2020 ◽

Author(s):

Lin Zhou ◽

Cheng Xing Yang ◽

Lin Chun Fang ◽

You Yuan Bao ◽

Zhi Gang Wang ◽

...

Keyword(s):

Signaling Pathway ◽

Primary Cell Culture ◽

Specific Inhibitor ◽

Akt Signaling ◽

Signal Pathway ◽

Cell Counting ◽

Migration And Invasion ◽

Specific Cell ◽

Primary Cells ◽

Akt Signaling Pathway

Abstract ObjectiveCraniopharyngiomas are rare histologically benign but clinically challenging neoplasms.The aim of this study is to explore the effect and significance of PI3K signal pathway on papillary craniopharyngioma cell growth and survival.MethodsWestern blotting (WB) was used to evaluate expression of the PI3K / AKT protein in Craniopharyngiomas tissues relative to health controls. Primary tumor cells were obtained from fresh papillary craniopharyngioma samples by primary cell culture and determined by cell morphology, immunofluorescence staining and specific cell markers expression. In this study, PCPs cell lines, isolated from fresh papillary craniopharyngioma samples, were treated with different concentrations of LY294002, a specific inhibitor of the PI3K / AKT signaling pathway, in order to evaluate their proliferation, migration and invasion. The proliferation effects was determined using Cell Counting Kit-8 and colony formation. Cell apoptosis and cell cycle were detected by flow cytometry. Furthermore, cell migration and invasion levels were detected by wound healing and Transwell assays, respectively.ResultsThe expression of p-PI3K and p-AKT were obviously higher in human craniopharyngioma tissue relative to their corresponding normal control. PCPs primary cells were isolated and detected by immunofluorescence, and PCPs cytokeratin (pan CK) and vimentin protein (VIM) were detected.The inhibition of PI3K / AKT signaling pathway can significantly inhibit the proliferation, migration and invasion of PCPs primary cells.ConclusionsLY294002 effectively inhibits the proliferation, migration and invasion of PCPs cells through the PI3K / AKT signaling pathway.

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Tizanidine hydrochloride exhibits a cytotoxic effect on osteosarcoma cells through the PI3K/AKT signaling pathway

Journal of International Medical Research ◽

10.1177/0300060519850395 ◽

2019 ◽

Vol 47 (8) ◽

pp. 3792-3802

Author(s):

Xue-Wu Xing ◽

Yu-Fu Sun ◽

Jun Zhao ◽

Zi-Xiang Pan ◽

Wen-Xue Jiang

Keyword(s):

Signaling Pathway ◽

Adrenergic Receptors ◽

Akt Signaling ◽

Cell Counting ◽

Migration And Invasion ◽

Cancer Cell Proliferation ◽

Akt Signaling Pathway ◽

Phosphatidylinositol 3 Kinase ◽

Expression Levels ◽

Underlying Mechanisms

Objectives α2-adrenergic receptors are reportedly involved in cancer cell proliferation, invasion, and apoptosis through regulation of diverse molecules, which implies that it contributes to tumor progression. However, the functional significance of α2-adrenergic receptors in osteosarcoma (OS) is unclear. Tizanidine hydrochloride (THC), an α2-adrenergic receptor agonist, is often used to alleviate symptoms of spasticity. This study investigated the functional implications of THC treatment on human OS cells and the underlying mechanisms of resulting changes. Methods The proliferation of U2 OS cells was assessed by Cell Counting Kit-8; the migration and invasion capacities of U2 OS cells were then analyzed by transwell assay. Moreover, apoptosis in U2 OS cells was evaluated by flow cytometry and western blot analyses. Additionally, expression levels of key proteins in the phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT) signaling pathway were measured. Results THC inhibited the proliferation, migration, and invasion of U2 OS cells, but promoted apoptosis within these cells. Expression levels of p-AKT, p-mTOR, and p-P70S6K were reduced by exposure to THC, suggesting involvement of PI3K/AKT signaling in THC-induced cytotoxicity within OS cells. Conclusions THC may play a novel role in OS cell cytotoxicity, and these findings suggest a potent therapeutic strategy for OS treatment.

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Silence of long non-coding RNA KCNQ1OT1 inhibits the proliferation, migration and invasion of hepatocellular carcinoma cells through PI3K/AKT signaling pathway

10.21203/rs.3.rs-21344/v1 ◽

2020 ◽

Author(s):

Hongliang Mei ◽

Zhiguo Yu ◽

Guanqi Zhang ◽

Zhiyuan Huang ◽

Hanjun Li ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Signaling Pathway ◽

Cell Lines ◽

Negative Impact ◽

Akt Signaling ◽

Cell Counting ◽

Migration And Invasion ◽

Akt Signaling Pathway ◽

Potential Biomarker

Abstract Background: KCNQ1 opposite strand/antisense transcript 1 (KCNQ1OT1) has been reported to be associated with hepatocellular carcinoma (HCC), which is considered as one of the most common cancers worldwide. However, the mechanism of action of KCNQ1OT1 in human HCC has not been fully explained. In this study, we aimed to explore the functional role and the potential mechanism of KCNQ1OT1 in human HCC.Methods: First, we analyzed the expression levels of KCNQ1OT1 in HCC tissues in starBase database and detected the expression of KCNQ1OT1 in HCC cell lines by quantitative real-time polymerase chain reaction assays. Next, we analyzed the role of KCNQ1OT1 in migration, invasion and proliferation of HCC by scratch wound healing, transwell and cell counting kit-8 assays. Finally, we analyzed the potential interrelationship between KCNQ1OT1 and PI3K/AKT signaling pathway through western blot assays.Results: Based on bioinformatics analyses, we found that KCNQ1OT1 was highly expressed in HCC tissues and its high expression was associated with a poor prognosis in HCC patients. We also confirmed an abnormal increase in the expression of KCNQ1OT1 in HCC cell lines. KCNQ1OT1 knockdown was found to have a negative impact on proliferation, migration and invasion of HCC cells. In addition, interference with the expression of KCNQ1OT1 reduced the phosphorylation level of AKT and the protein level of PI3K, indicating the association of KCNQ1OT1 with the PI3K/AKT signaling pathway.Conclusions: Collectively, this study confirmed the important role of KCNQ1OT1 in promoting HCC growth and revealed the inhibitory effect of KCNQ1OT1 on the PI3K/AKT signaling pathway. This work may contribute to a better understanding of HCC progression and provide a potential biomarker for HCC.

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Vincosamide inhibits malignant behaviors of hepatocellular carcinoma cells by activating caspase-3 activity and blocking the PI3K/AKT signaling pathway

10.21203/rs.3.rs-86111/v1 ◽

2020 ◽

Author(s):

Mingyue Zhu ◽

Haipeng Feng ◽

Bo Lin ◽

Ying Zhou ◽

Yifeng Zheng ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Signaling Pathway ◽

Caspase 3 ◽

Akt Signaling ◽

Cell Counting ◽

Migration And Invasion ◽

Mitochondrial Morphology ◽

Akt Signaling Pathway ◽

Related Factors ◽

Hcc Cells

Abstract Background: Vincosamide(Vinco) was first identified in the methanolic extract of the leaves of Psychotria leiocarpa, and Vinco has important anti-inflammatory effects and activity against cholinesterase, Vinco also has a trait to ant-tumor. However, whether Vinco is able to inhibit the malignant behaviors of hepatocellular carcinoma(HCC) cells is still unclear. In the present study, we explored the role of Vinco in suppressing the malignant behaviors of HCC cells.Methods: MTT, trypan blue exclusion assay and the Cell Counting Kit(CCK)-8 analysis were applied to detect the proliferation and death of HCC cells; electron microscopy was performed to observe the change of cellular mitochondrial morphology; scratch repair and Transwell assays were used to analyze the migration and invasion of HCC cells; the expression and localization of proteins were detected by laser confocal microscopy and Western blotting; and the growth of the cancer cells in in vivo was assessed in a mouse tumorous model.Results: At a dose of 10-80 mg/ml, Vinco inhibited the proliferation and promoted death of HCC cells in a dose-independent manner, but had low cytotoxcity effect on normal liver cells. Additionally, 80 mg/ml of Vinco could significantly disrupt the morphology of mitochondria, suppress the migration and invasion of HCC cells. The growth of HCC cells in the animal tumorous model was significantly inhibited after treatment with Vinco (10 mg/kg/day) for 3 days. The results of the present study indicated that Vinco (10-80 mg/ml) played a role in activating caspase-3, promoting the expression of PTEN, and inhibiting the phosphorylation of AKT(Ser473) and mTOR(Thr2448), Vinco also has a trait for suppressing the expression of CXCR4, Src, MMP9, EpCAM, Ras and Oct4 in HCC cells.Conclusions: Vinco has a role in inhibiting the malignant behaviors of HCC cells; the role molecular mechanism of Vinco maybe involve in restraining expression of the growth-, metastasis-related factors Src, Ras, MMP9, EpCAM and CXCR4, and activating the activity of caspase-3. Vinco also could block PI3K/AKT signaling pathway. Thus, Vinco is an available chemotherapy for HCC patients.

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Long non-coding RNA HULC regulates growth and metastasis of human glioma cells via induction of apoptosis and inhibits cell migration and invasion

Archives of Medical Science ◽

10.5114/aoms.2020.100834 ◽

2020 ◽

Author(s):

Junfeng Ma ◽

Liang Zhou

Keyword(s):

Cancer Cells ◽

Signaling Pathway ◽

Glioma Cells ◽

Akt Signaling ◽

Cell Counting ◽

Migration And Invasion ◽

Human Glioma ◽

Akt Signaling Pathway ◽

Non Coding Rna ◽

Long Non Coding Rna

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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