Notch1 Signaling Modulates Hypoxia-induced Multidrug Resistance of Human Laryngeal Cancer Cells

Abstract Background: Laryngeal carcinoma is one of the common malignant tumors of the head and neck. Multidrug resistance (MDR) remains a critical problem in the chemotherapy for patients with laryngeal cancer. This study aims to clarify the role and mechanisms of Notch1 signaling on MDR induced by hypoxia in laryngeal cancer cells.Methods and Results: Laryngeal carcinoma cells were cultured under normoxia or hypoxia. Notch1 expression was inhibited by small interfering RNA (siRNA). The expression of Notch1, Hes1, Hey1, MDR1 and survivin mRNA was determined by Real-time PCR. The expression of Notch1, Notch1 intracellular domain (N1ICD), MDR1/P-gp and survivin protein was detected by Western blot. Current research showed that hypoxia could upregulate Notch1 expression and the activity of Notch1 signaling. Furthermore, suppression of Notch1 expression could effectively down-regulate the activity of Notch1 signaling and the expression of MDR and survivin genes in laryngeal cancer cells under hypoxia (P<0.05). Cell Counting Kit-8 (CCK-8) assay confirmed that the sensitivity of hypoxic laryngeal cancer cells to a variety of drugs could be up-regulated by suppressing Notch1 expression (P<0.05). Additionally, flow cytometry (FCM) showed that suppression of Notch1 expression significantly increased cisplatin-induced apoptosis and intracellular Rh123 (Rh123) accumulation in hypoxic laryngeal carcinoma cells (P<0.05). Conclusions: Notch1 signalling could be regarded as a pivotal regulator for mediating hypoxia-induced MDR in laryngeal cancer cells by regulating survivin-mediated apoptosis resistance and MDR1/P-gp-mediated drug transport.

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Effect of Notch1 Signaling on Cellular Proliferation and Apoptosis of Human Laryngeal Carcinoma

10.21203/rs.3.rs-1101111/v1 ◽

2021 ◽

Author(s):

Dawei Li ◽

Yifei Zhang ◽

Penghui Chen ◽

Jin Xie ◽

Dan Xu

Keyword(s):

Cell Proliferation ◽

Cancer Cells ◽

Laryngeal Cancer ◽

Cell Apoptosis ◽

Laryngeal Carcinoma ◽

Apoptosis Resistance ◽

Neoplastic Cells ◽

Notch1 Signaling ◽

Proliferation And Apoptosis ◽

Signaling Activity

Abstract The pathological processes of occurrence and development of malignancies include the excessive proliferation and apoptosis resistance of neoplastic cells. The study aims to identify the effects of Notch1 signaling on the proliferation and apoptosis of laryngeal cancer cells in hypoxic microenvironment. Notch1 and Ki-67 expression in laryngeal squamous cell carcinoma (LSCC) tissue samples were detected by immunohistochemistry. The apoptotic index (AI) of LSCC was evaluated by TUNEL method. In laryngeal cancer cells, small interfering RNA (siRNA) technology was to inhibit Notch1 expression. Meanwhile, Real-time PCR detected Notch1, Hes1 and Hey1 mRNA expression, and Western blot detected Notch1 and Notch1 intracellular domain (N1ICD) protein expression. Annexin V-FITC/propidium iodide staining and Cell Counting Kit‑8 methods measured cell apoptosis and proliferation, respectively. Notch1 expression was detected in 63.55％(68/107) of LSCC samples and was significantly related to the proliferation index (PI) (P < 0.05) and AI (P < 0.05) in LSCC tissues. Furthermore, it was confirmed that hypoxia could induce proliferation and inhibit apoptosis of laryngeal carcinoma cells (P < 0.05). Meanwhile, Notch1 expression and Notch1 signaling activity could be upregulated by hypoxia (P < 0.05). In contrast, suppression of Notch1 signaling activity in hypoxic neoplastic cells could obviously decrease cell proliferation and increase cell apoptosis (both P<0.05). Our study has demonstrated that hypoxia may promote cell proliferation and inhibit cell apoptosis of laryngeal carcinoma. Notch1 signalling may exert a pivotal role in regulating the proliferation and apoptosis resistance of laryngeal cancer cells under hypoxia.

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Effect and Mechanism of Survivin on Hypoxia-Induced Multidrug Resistance of Human Laryngeal Carcinoma Cells

BioMed Research International ◽

10.1155/2019/5696801 ◽

2019 ◽

Vol 2019 ◽

pp. 1-6

Author(s):

Dan Xu ◽

Da Wei Li ◽

Jin Xie ◽

Xin Wei Chen

Keyword(s):

Multidrug Resistance ◽

Rna Interference ◽

Cancer Cells ◽

Propidium Iodide ◽

Laryngeal Carcinoma ◽

Survivin Expression ◽

Annexin V ◽

Carcinoma Cells ◽

Propidium Iodide Staining ◽

Human Laryngeal Carcinoma

This study aimed at clarifying the mechanism and role of survivin in hypoxia-induced multidrug resistance (MDR) of laryngeal carcinoma cells. Human laryngeal cancer cells were incubated under hypoxia or normoxia. The expression of survivin was silenced by performing RNA interference. Additionally, by Western blot and real-time quantitative RT-PCR, survivin expression was detected. The sensitivity of human laryngeal carcinoma cells to multiple drugs was measured by CCK-8 assay. Meanwhile, the apoptosis of cells induced by cisplatin or paclitaxel was assessed by Annexin-V/propidium iodide staining analysis. Under hypoxic conditions, the upregulation of survivin was abolished by RNA interference. Then, CCK-8 analysis demonstrated that the sensitivity to multiple agents of laryngeal carcinoma cells could be increased by inhibiting survivin expression (P<0.05). Moreover, Annexin-V/propidium iodide staining analysis revealed that decreased expression of survivin could evidently increase the apoptosis rate of laryngeal carcinoma cells that were induced by cisplatin or paclitaxel evidently (P<0.05). Our data suggests that hypoxia-elicited survivin may exert a pivotal role in regulating hypoxia-induced MDR of laryngeal cancer cells by preventing the apoptosis of cells induced by chemotherapeutic drug. Thus, blocking survivin expression in human laryngeal carcinoma cells may provide an avenue for gene therapy.

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Cytotoxicity, Oxidative Stress, Cell Cycle Arrest, and Mitochondrial Apoptosis after Combined Treatment of Hepatocarcinoma Cells with Maleic Anhydride Derivatives and Quercetin

Oxidative Medicine and Cellular Longevity ◽

10.1155/2017/2734976 ◽

2017 ◽

Vol 2017 ◽

pp. 1-16 ◽

Cited By ~ 16

Author(s):

Gabriela Carrasco-Torres ◽

Rafael Baltiérrez-Hoyos ◽

Erik Andrade-Jorge ◽

Saúl Villa-Treviño ◽

José Guadalupe Trujillo-Ferrara ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Cell Cycle ◽

Maleic Anhydride ◽

Cancer Cells ◽

Combination Treatment ◽

Malignant Tumors ◽

Combined Treatment ◽

Current Data ◽

Carcinoma Cells ◽

Hepatocellular Carcinoma Cells

The inflammatory condition of malignant tumors continually exposes cancer cells to reactive oxygen species, an oxidizing condition that leads to the activation of the antioxidant defense system. A similar activation occurs with glutathione production. This oxidant condition enables tumor cells to maintain the energy required for growth, proliferation, and evasion of cell death. The objective of the present study was to determine the effect on hepatocellular carcinoma cells of a combination treatment with maleic anhydride derivatives (prooxidants) and quercetin (an antioxidant). The results show that the combination of a prooxidant/antioxidant had a cytotoxic effect on HuH7 and HepG2 liver cancer cells, but not on either of two normal human epithelial cell lines or on primary hepatocytes. The combination treatment triggered apoptosis in hepatocellular carcinoma cells by activating the intrinsic pathway and causing S phase arrest during cell cycle progression. There is also clear evidence of a modification in cytoskeletal actin and nucleus morphology at 24 and 48 h posttreatment. Thus, the current data suggest that the combination of two anticarcinogenic drugs, a prooxidant followed by an antioxidant, can be further explored for antitumor potential as a new treatment strategy.

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Decrease in Multidrug Resistance-associated Protein 2 Activities by Knockdown of Phosphatidylinositol 4-phosphate 5-kinase in Hepatocytes and Cancer Cells

Journal of Pharmacy & Pharmaceutical Sciences ◽

10.18433/jpps30444 ◽

2019 ◽

Vol 22 ◽

pp. 576-584 ◽

Cited By ~ 1

Author(s):

Atsushi Kawase ◽

Yuta Inoue ◽

Miho Hirosoko ◽

Yuka Sugihara ◽

Hiroaki Shimada ◽

...

Keyword(s):

Plasma Membrane ◽

Multidrug Resistance ◽

Cancer Cells ◽

Hepg2 Cells ◽

Carcinoma Cells ◽

Efflux Transporters ◽

Membrane Localization ◽

Intracellular Accumulation ◽

Primary Hepatocytes ◽

Phosphatidylinositol 4

Purpose: The plasma membrane localization and transport activity of multidrug resistance-associated protein 2 (MRP2/ABCC2) and P-glycoprotein (P-gp/ABCB1) efflux transporters are governed by transporter-associated proteins. Phosphatidylinositol 4,5-bisphosphate (PIP2) formed by phosphatidylinositol 4-phosphate 5-kinase type 1 (PIP5K1) activates the linker function of radixin for efflux transporters. Radixin is involved in the plasma membrane localization of efflux transporters. We examined whether PIP5K1 could be a target for the modulation of transporter activities in hepatocytes and cancer cells. Methods: The effects of PIP5K1 depletion by siRNA in mouse primary hepatocytes, PANC1 human pancreatic carcinoma cells, and HepG2 human hepatocellular carcinoma cells on the intracellular accumulation of MRP2 and P-gp substrates were examined. Results: PIP5K1A depletion resulted in increased intracellular accumulation of carboxydichlorofluorescein, a MRP2 fluorescent substrate, in mouse primary hepatocytes, PANC1 cells, and HepG2 cells. In PANC1 and HepG2 cells, the transport activities of MRP2 were significantly decreased by PIP5K1C depletion. However, the transport activities of P-gp were unchanged by PIP5K1 depletion. PIP2 levels were unchanged between control and PIP5K1A- or PIP5K1C-depleted HepG2 cells. MRP2 mRNA levels showed few changes in HepG2 cells following PIP5K1A or PIP5K1C depletion. The expression of phosphorylated radixin was decreased by PIP5K1A and PIP5K1C depletion, although total radixin levels were unchanged. Conclusions: These data suggest that PIP5K1A and PIP5K1C could be target proteins for modulating MRP2 function, partly because of the resulting changes of the linker function of radixin.

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Trop2 Promotes Multidrug Resistance by Regulating Notch1 Signaling Pathway in Gastric Cancer Cells

Medical Science Monitor ◽

10.12659/msm.919566 ◽

2020 ◽

Vol 26 ◽

Cited By ~ 4

Author(s):

Xingwang Kuai ◽

Lizhou Jia ◽

Tingting Yang ◽

Xiaochen Huang ◽

Wei Zhao ◽

...

Keyword(s):

Gastric Cancer ◽

Multidrug Resistance ◽

Cancer Cells ◽

Signaling Pathway ◽

Gastric Cancer Cells ◽

Notch1 Signaling ◽

Notch1 Signaling Pathway

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The expression and significance of AKR1B10 in laryngeal squamous cell carcinoma

Scientific Reports ◽

10.1038/s41598-021-97648-y ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Jixuan Liu ◽

Hongyan Ban ◽

Yafang Liu ◽

Jinsong Ni

Keyword(s):

Laryngeal Cancer ◽

Laryngeal Carcinoma ◽

Nicotinamide Adenine Dinucleotide Phosphate ◽

Lipid Synthesis ◽

Prognostic Indicator ◽

Carcinoma Cells ◽

Matrix Metalloproteinase 2 ◽

Migration And Invasion ◽

Ki 67 ◽

Degree Of Differentiation

AbstractAldosterone reductase family 1 member B10 (AKR1B10) is a nicotinamide adenine dinucleotide phosphate (reduced coenzyme II)-dependent oxidoreductase, and its biological functions include carbonyl detoxification, hormone metabolism, osmotic adjustment, and lipid synthesis. Studies suggested that AKR1B10 is a new biomarker for cancer based on its overexpression in epithelial tumors, such as breast cancer, cervical cancer, and lung cancer. At present, studies on the expression of AKR1B10 in laryngeal cancer have not been reported. However, we found that AKR1B10 is upregulated in laryngeal carcinoma, and its expression was negatively correlated with the degree of differentiation. In addition, AKR1B10 expression was positively correlated with tumor size; lymph node metastasis; alcohol use; and Ki-67, mutant p53, and matrix metalloproteinase 2 expression. AKR1B10 was overexpressed in Hep-2 laryngeal carcinoma cells. Oleanolic acid inhibited AKR1B10 activity and expression in Hep-2 cells and suppressed Hep-2 cell proliferation, migration, and invasion. Therefore, AKR1B10 may be related to the development of laryngeal carcinoma, suggesting its use as a prognostic indicator for laryngeal cancer.

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Tumor-suppressor miRNA-27b-5p regulates the growth and metastatic behaviors of ovarian carcinoma cells by targeting CXCL1

10.21203/rs.3.rs-20826/v2 ◽

2020 ◽

Author(s):

Chun Hua Liu ◽

Xue Ning Jing ◽

Xiao Lan Liu ◽

Shan Yong Qin ◽

Min Wei Liu ◽

...

Keyword(s):

Ovarian Cancer ◽

Ovarian Carcinoma ◽

Cancer Cells ◽

Carcinoma Cells ◽

Cell Counting ◽

Luciferase Reporter ◽

Ovarian Cancer Cells ◽

Skov3 Cell ◽

Ovarian Carcinoma Cells ◽

The Impact

Abstract BackgroundMicroRNAs (miRNAs) play crucial functions in the progression of ovarian cancer. MicroRNA-27b-5p (miR-27b-5p) has been identified as a cancer-associated miRNA. Nevertheless, the expression profile of miR-27b-5p and its functions in ovarian cancer are unexplored.MethodsqRT-PCR and western blot analysis were used to detect the levels of miR-27b-5p and C-X-C motif chemokine ligand 1 (CXCL1). The impact of miR-27b-5p on ovarian cancer cells proliferation, migration and invasion in vitro were investigated using Cell Counting Kit-8 (CCK8), wound healing and Transwell, respectively. The expression of matrix metalloprotein-2/9 (MMP-2/9) were measured using immunofluorescence staining. Bioinformatics and luciferase reporter analysis were used to predict the target of miR-27b-5p. The growth of ovarian cancer cells in vivo was evaluated using transplanted tumor model.ResultsHere, we demonstrated that miR-27b-5p was downregulated in ovarian carcinoma cells and clinical specimens. Higher expression of miR-27b-5p was associated with an unfavorable overall survival in patients with ovarian cancer. Upregulation of miR-27b-5p decreased the viability, migration ability and invasion capacity of SKOV3 and OVCAR3 cell. MiR-27b-5p also inhibited the growth of SKOV3 cell in nude mice. Additionally, we verified that CXCL1 was a target of miR-27b-5p in ovarian carcinoma cells. Restoring the expression of CXCL1 abolished the inhibitory impacts of miR-27b-5p in ovarian cancer carcinoma cells.ConclusionThis research revealed that miR-27b-5p restrained the progression of ovarian carcinoma possibly via targeting CXCL1.

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Overexpressed active Notch1 induces cell growth arrest of HeLa cervical carcinoma cells

International Journal of Gynecological Cancer ◽

10.1111/j.1525-1438.2007.00927.x ◽

2007 ◽

Vol 17 (6) ◽

pp. 1283-1292 ◽

Cited By ~ 28

Author(s):

L. Wang ◽

H. Qin ◽

B. Chen ◽

X. Xin ◽

J. Li ◽

...

Keyword(s):

Cell Growth ◽

Notch Signaling ◽

Cervical Carcinoma ◽

Molecular Mechanisms ◽

Malignant Tumors ◽

Critical Role ◽

Carcinoma Cells ◽

Human Cervical Carcinoma ◽

Notch1 Signaling ◽

Context Specific

Human cervical carcinoma is one of the most common malignant tumors, but the mechanisms that orchestrate the multiple oncogenic insults required for initiation and progression are not clear. Notch signaling plays a critical role in maintaining the balance between cell proliferation, differentiation, and apoptosis, but perturbed Notch signaling may contribute to tumorigenesis. We now show that Notch1 is detected in all cervical cancer, including advanced diseases. We also constitutively overexpressed active Notch1 in human cervical carcinoma to explore the effects of Notch1 signaling on human cervical carcinoma cell growth and to investigate the underlying molecular mechanisms. The signaling may participate in the development of human cervical carcinoma cells, but overexpressed active Notch1 inhibits their growth through induction of cell cycle arrest. Increased Notch1 signaling induced a downmodulation of human papillomavirus transcription through suppression of activator protein (AP)-1 activity by upregulation of c-Jun and the decreased expression of c-Fos. Thus, Notch1 signaling plays a key role and exerts dual effects, functioning in context-specific manner

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