LHPP suppresses proliferation, migration, and invasion and promotes apoptosis in pancreatic cancer

Abstract Pancreatic cancer (PaCa) is a common malignant tumor of the digestive system with poor prognosis and no ideal treatment for inoperable patients, which is partly due to delayed diagnoses. It is recently reported that the protein histidine phosphatase LHPP is a tumor suppressor in hepatocellular carcinoma, cervical cancer, and bladder cancer. So far, there is no study on the expression level of LHPP in PaCa, and its mechanism of action on tumors is unclear. In this experiment, LHPP expression was lower in cancer tissues than that in normal pancreatic tissue, and clinicopathological results showed that LHPP expression was correlated with the degree of differentiation and lymphatic metastasis of pancreatic carcinoma. The biological characteristics of LHPP in PaCa cells were examined by the cell counting kit-8 assay, transwell assay, and monoclonal formation test. The inhibitory mechanism of LHPP in PaCa cells was determined using Western blotting and flow cytometry. The results showed that LHPP restrained PaCa cell proliferation, migration, and invasion. Increased LHPP expression promoted the apoptosis of PaCa cells through higher activation of cleaved-PARP and cleaved-Casp3 and lower activation of cIAP1. Importantly, the increase in LHPP enhanced PTEN expression and decreased the phosphorylated AKT level. Moreover, LHPP-induced apoptosis was diminished by SC79 (AKT activator) in PaCa cells. In conclusion, LHPP blocks proliferation, migration, and invasion and enhances apoptosis in PaCa cells through the PTEN/AKT signaling pathway.

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Association of γ-aminobutyric acid type A receptor-associated protein with prognosis in patients after radical pancreatic cancer treatment

10.21203/rs.3.rs-51240/v2 ◽

2020 ◽

Author(s):

Chao Zhang ◽

Wenhao Tang ◽

Yanyuan Tu ◽

Zurong Yuan ◽

Wei Wang ◽

...

Keyword(s):

Pancreatic Cancer ◽

Adjuvant Chemotherapy ◽

Cancer Patients ◽

Cox Regression ◽

Pancreatic Tissue ◽

Postoperative Adjuvant Chemotherapy ◽

Normal Pancreatic Tissue ◽

Free Survival ◽

Patient Prognosis ◽

Cancer Tissues

Abstract Background：Pancreatic cancer is difficult to cure and many factors influence patient prognosis, of which autophagy is a recent research hotspot, and GABARAP plays a key role in autophagy, which has been found to interfere with cancer cell survival and metastasis in a variety of tumours. In this study, we analysed the correlation between GABARAP and patient prognosis in pancreatic cancer, as well as its correlation with clinicopathological parameters and its impact on the efficacy of chemotherapy in pancreatic cancer patients.Methods: The pancreatic tissues of 76 pancreatic cancer patients after R0 resection were screened according to the criteria, and the expression levels of GABARAP were determined by using immunohistochemistry (MaxVision) to label the pancreatic cancer tissues and normal pancreatic tissue around carcinoma in these two types of specimens, and the relationship between GABARAP and other factors and the disease-free and overall survival of patients after radical pancreatic cancer treatment was evaluated by single factor survival analysis and Cox regression analysis.Results：The expression ratio of GABARAP in pancreatic cancer tissue was drastically higher than that in normal pancreatic tissue adjacent to cancer. Cox regression model evaluation showed that GABARAP and postoperative adjuvant chemotherapy were the overall survival of patients after radical pancreatic cancer Period independent prognostic indicators and both are protective factors for the prognosis of pancreatic cancer patients. Further data analysis found that postoperative chemotherapy drastically increased the total patient Survival period in GABARAP-negative patients but had no drastically effect on the patient's relapse-free survival period.Conclusion: The expression of GABARAP in pancreatic cancer tissues was drastically up-regulated, and patients with high expression of GABARAP in pancreatic cancer tissues had better prognosis, but had no drastically effect on the relapse-free survival of patients after radical operation of pancreatic cancer. The expression of GABARAP in pancreatic cancer tissues and Postoperative adjuvant chemotherapy is an independent indicator of patients' prognosis after radical pancreatic cancer resection. Both are protective factors. The high expression of GABARAP in pancreatic cancer may indicate that the adjuvant chemotherapy is low benefit.

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

10.21203/rs.3.rs-119581/v1 ◽

2020 ◽

Author(s):

Lin Zhou ◽

Cheng Xing Yang ◽

Lin Chun Fang ◽

You Yuan Bao ◽

Zhi Gang Wang ◽

...

Keyword(s):

Signaling Pathway ◽

Akt Signaling ◽

Cell Counting ◽

Migration And Invasion ◽

Specific Cell ◽

Primary Cells ◽

Akt Signaling Pathway ◽

Phosphatidylinositol 3 Kinase ◽

Growth And Survival ◽

Pi3k Signaling Pathway

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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Overexpression of miR-142-5p suppresses the progression of cervical cancer through targeting PIK3AP1 expression

Molecular and Cellular Biology ◽

10.1128/mcb.00363-20 ◽

2020 ◽

Author(s):

Junliang Guo ◽

Tian Tang ◽

Jinhong Li ◽

Yihong Yang ◽

Yi Quan ◽

...

Keyword(s):

Cervical Cancer ◽

Cancer Cells ◽

Signaling Pathway ◽

Akt Signaling ◽

Migration And Invasion ◽

Akt Signaling Pathway ◽

Gain Function ◽

Cervical Cancer Cells ◽

Target Binding ◽

Cancer Tissues

The aim of current study was to explore the mechanism of miR-142-5p in cervical cancer through mediating the PIK3AP1/P13K/AKT axis. To this end, RT-qPCR and Western blot analysis results revealed that miR-142-5p was poorly expressed, whereas PIK3AP1 was highly expressed in cervical cancer tissues and cells. Furthermore, miR-142-5p was hypermethylated in cervical cancer, as reflected by MS-PCR and ChIP assessment of enrichment of DNMT1/DNMT3a/DNMT3b in the promoter region of miR-142-5p. A target binding relationship between miR-142-5p and PIK3AP1 was established, showing that miR-142-5p targeted and inhibited the expression of PIK3AP1. Loss- and gain- function assays were conducted to determine the roles of miR-142-5p and PIK3AP1 in cervical cancer cells. CCK-8, flow cytometry and Transwell assay results revealed that overexpression of miR-142-5p in cervical cancer cells downregulated PIK3AP1 and inhibited the P13K/AKT signaling pathway, leading to reduced proliferation, migration, and invasion capacity of cervical cancer cells, but enhanced apoptosis. Collectively, epigenetic regulation of miR-142-5p targeted PIK3AP1 to inactivate the P13K/AKT signaling pathway, thus suppressing development of cervical cancer, which presents new targets for the treatment of cervical cancer.

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ALKBH5 Inhibits Pancreatic Cancer Motility by Decreasing Long Non-Coding RNA KCNK15-AS1 Methylation

Cellular Physiology and Biochemistry ◽

10.1159/000491915 ◽

2018 ◽

Vol 48 (2) ◽

pp. 838-846 ◽

Cited By ~ 63

Author(s):

Yuan He ◽

Hao Hu ◽

Yandong Wang ◽

Hao Yuan ◽

Zipeng Lu ◽

...

Keyword(s):

Pancreatic Cancer ◽

Cancer Cells ◽

Epithelial Mesenchymal Transition ◽

The Body ◽

Migration And Invasion ◽

Mesenchymal Transition ◽

Cancer Pathogenesis ◽

Non Coding Rna ◽

Cancer Tissues ◽

Long Non Coding Rna

Background/Aims: Mounting evidence suggests that epitranscriptional modifications regulate multiple cellular processes. N6-Methyladenosine (m6A), the most abundant reversible methylation of mRNA, has critical roles in cancer pathogenesis. However, the mechanisms and functions of long non-coding RNA (lncRNA) methylation remain unclear. Pancreatic cancer resulted in 411,600 deaths globally in 2015. By the time of pancreatic cancer diagnosis, metastasis has often occurred in other parts of the body. The present study sought to investigate lncRNA m6A modification and its roles in pancreatic cancer. Methods: Differential expression between cancer cells and matched normal cells was evaluated to identify candidate lncRNAs. The lncRNA KCNK15-AS1 was detected in cancer tissues and various pancreatic cells using RT-qPCR. KCNK15-AS1 was transfected into cells to explore its role in migration and invasion. Then, m6A RNA immunoprecipitation was performed to detect methylated KCNK15-AS1 in tissues and cells. Epithelial–mesenchymal transition (EMT) markers were used to evaluate KCNK15-AS1-mediated EMT processes. Results: KCNK15-AS1 was downregulated in pancreatic cancer tissues compared with paired adjacent normal tissues. KCNK15-AS1 inhibited migration and invasion in MIA PaCa-2 and BxPC-3 cells. Furthermore, total RNA methylation in cancer cells was significantly enriched relative to that in immortalized human pancreatic duct epithelial (HPDE6-C7) cells. In addition, the m6A eraser ALKBH5 was downregulated in cancer cells, which can demethylate KCNK15-AS1 and regulate KCNK15-AS1-mediated cell motility. Conclusion: Our results have revealed a novel mechanism by which ALKBH5 inhibits pancreatic cancer motility by demethylating lncRNA KCNK15-AS1, identifying a potential therapeutic target for pancreatic cancer.

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Downregulation of cAMP-Dependent Protein Kinase Inhibitor-b Promotes Preeclampsia by Decreasing Phosphorylated Akt

Reproductive Sciences ◽

10.1007/s43032-020-00258-8 ◽

2020 ◽

Vol 28 (1) ◽

pp. 178-185

Author(s):

Chunfeng Liu ◽

Hao Wang ◽

Mo Yang ◽

Yiheng Liang ◽

Li Jiang ◽

...

Keyword(s):

Protein Kinase ◽

Matrix Metalloproteinase ◽

Signaling Pathway ◽

Kinase Inhibitor ◽

Akt Signaling ◽

Migration And Invasion ◽

Akt Signaling Pathway ◽

Dependent Protein Kinase ◽

Phosphorylated Akt ◽

Dependent Protein

AbstractPreeclampsia is a multi-system disease that is unique to human pregnancy. Impaired extravillous trophoblast migration and invasion accompanied by poor spiral vascular remodeling is thought to be the initial reason. This study investigated cAMP-dependent protein kinase inhibitor-b(PKIB) expression in placentas and its involvement in the pathogenesis of PE. We used immunohistochemistry and western blotting to calculate PKIB levels in the placentas. Then we knocked down PKIB by siRNA and used real-time cell analysis to assess the invasion and migration ability of trophoblasts. Tube formation assay and spheroid sprouting assay were utilized to identify the ability to form vessels of trophoblasts. At last, western blotting was used to demonstrate the level of phosphorylated Akt, as well as downstream-related genes of Akt signaling pathway in trophoblasts. We first found that PKIB expression level was lower in the PE placentas than in the normal placentas. In addition, we found that downregulation of PKIB can inhibit the migration, invasion, and the ability to form vessels of HTR8/SVneo cells. Downregulation of PKIB leaded to a decrease in phosphorylated Akt, as well as downstream proteins such as matrix metalloproteinase 2, matrix metalloproteinase 9, and glycogen synthase kinase 3β, which are related to migration and invasion. Our study revealed that the downregulation of PKIB expression resulted in decreased migration, invasion, and vessel formation ability by regulating Akt signaling pathway in placental trophoblasts in PE.

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TM4SF1 Regulates Pancreatic Cancer Migration and Invasion In Vitro and In Vivo

Cellular Physiology and Biochemistry ◽

10.1159/000445664 ◽

2016 ◽

Vol 39 (2) ◽

pp. 740-750 ◽

Cited By ~ 19

Author(s):

Jia Cao ◽

Jia-chun Yang ◽

Vijaya Ramachandran ◽

Thiruvengadam Arumugam ◽

De-feng Deng ◽

...

Keyword(s):

Pancreatic Cancer ◽

Pancreatic Tumor ◽

Migration And Invasion ◽

Qrt Pcr ◽

Pancreatic Cancer Cells ◽

Cancer Tissues ◽

Immunohistochemical Analyses ◽

Expression And Activity

Background/Aims: The cell surface protein transmembrane 4 L6 family member 1 (TM4SF1) has been detected in various tumors and plays a major role in the development of cancer. We aimed to investigate the effects of TM4SF1 on the migration and invasion of pancreatic cancer in vitro and in vivo and explore its related molecular mechanisms. Methods: qRT-PCR and immunohistochemical analyses were used to measure the expression of TM4SF1 in pancreatic cancer tissues and adjacent tissues. TM4SF1 was silenced using siRNA and shRNA to investigate the role of this protein in the proliferation and metastasis of pancreatic cancer cells. MTS and Transwell assays were used to examine the effect of TM4SF1 on pancreatic cancer cell lines. The expression and activity of MMP-2 and MMP-9 were determined by qRT-PCR, western blots and gelatin zymography. In vivo, orthotopic pancreatic tumor models were used to examine the formation of metastasis. Results: qRT-PCR and immunohistochemical analyses showed that TM4SF1 was highly expressed in pancreatic cancer tissues compared with the adjacent tissues. In in vitro experiments the silencing of TM4SF1 reduced cell migration and invasion and down-regulated the expression and activity of MMP-2 and MMP-9. However, no significant difference in cell proliferation was detected after silencing TM4SF1. Additionally, knocking down TM4SF1 decreased the formation of lung and liver metastases in orthotopic pancreatic tumor models. Conclusion: Our results demonstrate that the expression of TM4SF1 is higher in pancreatic cancer tissues and pancreatic cancer cell lines than controls. Knockdown of TM4SF1 inhibited the migration and invasion of pancreatic cancer cells by regulating the expression and activity of MMP-2 and MMP-9, which suggests that TM4SF1 may play a significant role in metastasis in pancreatic cancer.

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Pharmacokinetic Analysis of Dynamic [18F]FAZA PET Imaging in Pancreatic Cancer Patient

10.21203/rs.3.rs-39968/v1 ◽

2020 ◽

Author(s):

Fiona Li ◽

Edward Taylor ◽

Ivan Yeung ◽

David Jaffray ◽

Ur Metser ◽

...

Keyword(s):

Pancreatic Cancer ◽

Kinetic Parameters ◽

Normal Tissue ◽

Pancreatic Tissue ◽

Distribution Volume ◽

Information Criteria ◽

Pharmacokinetic Analysis ◽

Time Activity ◽

Normal Pancreatic Tissue ◽

Pet Tracer

Abstract Purpose This study assessed the pharmacokinetics of the hypoxia PET tracer, [18F]fluoroazomycin arabinoside ([18F]FAZA), in pancreatic cancer (PCa) patients and determined the optimal kinetic parameters to distinguish cancerous from normal pancreatic tissue. Method Twenty patients with pancreatic ductal adenocarcinoma underwent dynamic [ 18 F]FAZA scans. The tissue time activity curve (TAC) was analyzed using graphical methods to determine reversibility of tracer binding and with standard compartment (S2TC) model and flow modified two tissue compartment (F2TC) model, developed to incorporate transit time of tracer through the blood vessel, to estimate the kinetic parameters. The optimal parameter set to distinguish hypoxic tumors from normal tissues was determined using logistic regression. Results Both graphical and kinetic model analysis indicated that tracer was reversibly bound. According to the Akaike Information Criteria, the F2TC model fitted the tumor TAC better than the S2TC model. Total distribution volume, V T , estimated by the F2TC model for both tumor and normal pancreatic tissue was not significant but that estimated by the S2TC model was significantly different from Logan graphical analysis. The extravascular distribution volume ( DV ) and tracer dissociation rate constant ( k 4 ) can classify hypoxic PCa from normal tissue with sensitivity of 95% and negative predictive value of 89% (P<0.01). Conclusions Kinetic analysis of dynamic [ 18 F]FAZA PET can distinguish PCa from normal tissue with high sensitivity. The reversibility of [ 18 F]FAZA binding in hypoxic cells could be due to glutathionylation of the nitroreductase reduced products and their subsequent efflux from same cells via the ATP mediated multidrug resistant protein (MRP-1) efflux pump.

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Downregulation of HBx Restrains Proliferation, Migration, and Invasion of HepG2 Cells

Analytical Cellular Pathology ◽

10.1155/2021/6615979 ◽

2021 ◽

Vol 2021 ◽

pp. 1-11

Author(s):

Chaoqun Huang ◽

Wei Liu ◽

Xiaochuan Zhao ◽

Libin Zhao ◽

Fuxiang Wang

Keyword(s):

Liver Cancer ◽

Cancer Progression ◽

Hepg2 Cells ◽

Quantitative Polymerase Chain Reaction ◽

Mechanistic Study ◽

Migration And Invasion ◽

Cancer Pathogenesis ◽

Induced Apoptosis ◽

Cancer Tissues

Liver cancer is a major contributor to cancer-related death with poor survival for sufferers. Meanwhile, Hepatic B virus X protein (HBx) and XB130 are likely to participate in the pathogenesis of liver cancer. However, the detailed mechanism of HBx/XB130 in liver cancer remains to be further investigated. Our study explored the effects of HBx/XB130 on liver cancer progression. HBx and XB130 expression was detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blot. Overexpression of HBx and XB130 was found in liver cancer tissues and cells. Mechanistic study revealed that HBx could bind to and positively regulate XB130 in HepG2 cells. Subsequently, HBx expression was knocked down, while XB130 was overexpressed in HepG2 cells in order to observe the specific role of HBx/XB130 in liver cancer in vitro. Results of CCK-8, Transwell, wound healing, and colony formation assays suggested that HBx could mediate biological function of HepG2 cells by activating the XB130-mediated PI3K/AKT pathway. In summary, our data illustrate that inhibition of HBx effectively suppressed proliferation and metastasis and induced apoptosis of liver cancer cells, which might be partially reversed by XB130. HBx and XB130 may be potential targets for liver cancer pathogenesis.

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Hypoxia-Related Gene FUT11 Promoted Pancreatic Cancer Progression Via Maintaining The Stabilization of PDK1

10.21203/rs.3.rs-152464/v2 ◽

2021 ◽

Author(s):

Wenpeng Cao ◽

Zhirui Zeng ◽

Runsang Pan ◽

Zhiwei He ◽

Hao Wu ◽

...

Keyword(s):

Pancreatic Cancer ◽

Cell Growth ◽

Western Blot ◽

Cancer Progression ◽

Pyruvate Dehydrogenase Kinase ◽

Cell Counting ◽

Luciferase Assay ◽

Migration And Invasion

Abstract Background: Hypoxia participated in the occurrence and development of pancreatic cancer (PC). However, genes associated with hypoxia respond and their regulated mechanism in PC cells were unclear. The current research was aimed to illuminate the role and hypoxia regulated mechanism of fucosyltransferase 11 (FUT11) in the progression of PC.Methods: After predicting FUT11 as a key hypoxia associated gene in PC using bioinformatics analysis. The expression of FUT11 in PC using quantitative real-time fluorescent PCR, western blot and immunohistochemistry. The effects of FUT11 on PC cells proliferation, migration and invasion under normoxia and hypoxia were detected using Cell Counting Kit 8, 5-ethynyl-2’-deoxyuridine assay, colony formation assay and transwell assay. Spleen capsule injected liver metastasis and subcutaneously injected model were performed to confirm the effects of FUT11 in vivo. Furthermore, western blot, luciferase assay and immunoprecipitation were performed to explore the regulated relationship among FUT11, hypoxia-inducible factor 1α (HIF1α) and pyruvate dehydrogenase kinase 1 (PDK1) in PC.Results: FUT11 was markedly increased of PC cells in hypoxia, up-regulated in the PC clinical tissues, and predicted a poor outcome. Inhibition of FUT11 reduced PC cell growth and mobility of PC cells under normoxia and hypoxia conditions in vitro, and growth and mobility in vivo. FUT11 bind with PDK1 and regulated the expression PDK1 under normoxia and hypoxia. FUT11 knockdown significantly increased the degradation rate of PDK1 under hypoxia, while treatment with MG132 can relieve the degradation of PDK1 induced by FUT11 knockdown. Overexpression of PDK1 in PC cells under hypoxia conditions reversed the suppressiv impacts of FUT11 knockdown on PC cell growth and mobility. In addition, HIF1α bound to the enhancer of FUT11 and increased its expression, as well as co-expressing with FUT11 in PC tissues. Furthermore, overexpress of FUT11 partially rescued the suppressiv effects of HIF1α knockdown on PC cell growth and mobility in hypoxia conditions.Conclusion: Our data further implicate that hypoxia-induced FUT11 in PC contributes to proliferation and metastasis by maintaining the stability of PDK1, and suggest FUT11 maybe a novel and effective target for treatment of pancreatic cancer.

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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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