Stable knockdown of ZBTB7A promotes cell proliferation and progression in nasopharyngeal carcinoma

2018 ◽  
Vol 104 (1) ◽  
pp. 37-42 ◽  
Author(s):  
Fei Liu ◽  
Fengzhu Tang ◽  
Jiao Lan ◽  
Wei Jiao ◽  
Yongfeng Si ◽  
...  
Keyword(s):  
Nasopharyngeal Carcinoma ◽  
Cell Lines ◽  
Xenograft Model ◽  
Short Hair ◽  
Chain Reaction ◽  
Focus Formation ◽  
Cell Vitality ◽  
Empty Plasmid ◽  
Oncogenic Pathways ◽  
Polymerase Chain

Aims: Although high expression of ZBTB7A is positively relative to metastasis in nasopharyngeal carcinoma (NPC) patients, the association between its low expression and metastasis of NPC remains unclear. The present study aimed to definitely identify the association. Methods: The level of ZBTB7A was effectively knocked down by stable transfection of short hair RNA plasmid in NPC cell lines CNE2 and 5-8F (shRNA-CNE2 and shRNA-5-8F), compared with the cells that stably transfected empty plasmid (NC-CNE2 and NC-5-8F). The levels of ZBTB7A were assessed by real-time polymerase chain reaction and Western blot in the cell lines. MTT assay, colorimetric focus-formation assay, flow cytometry, wound healing assay, transwell assays, and xenograft model were performed to analyze cell vitality, proliferation, cell cycle, migration, invasion, and tumorigenicity. Results: The levels of ZBTB7A were effectively reduced in shRNA-CNE2 and shRNA-5-8F. Their carcinogenicity was stronger separately than the abilities of NC-CNE2 and NC-5-8F. NC-CNE2 and shRNA-CNE2 were selected to establish the xenograft model because of their stronger tumorigenicity than NC-6-10B and shRNA-5-8F. The assay showed that shRNA-CNE2 had stronger tumorigenicity than NC-CNE2. Conclusions: The results demonstrated the reverse association between the expression of ZBTB7A and the tumorigenicity of NPC. We postulate that some oncogenic pathways, which are suppressed by ZBTB7A, will vicariously promote the proliferation and progression of NPC when ZBTB7A is decreased.

Expression of Platelet Membrane Glycoprotein V in Human Megakaryocytes and Megakaryocytic Cell Lines: A Study Using a Novel Monoclonal Antibody against GPV

1994 ◽  
Vol 72 (05) ◽  
pp. 762-769 ◽  
Author(s):  
Toshiro Takafuta ◽  
Kingo Fujirmura ◽  
Hironori Kawano ◽  
Masaaki Noda ◽  
Tetsuro Fujimoto ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cell Lines ◽  
Immunohistochemical Analysis ◽  
Specific Protein ◽  
Platelet Membrane ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Flow Cytometric ◽  
Polymerase Chain ◽  
Megakaryocytic Cell

SummaryGlycoprotein V (GPV) is a platelet membrane protein with a molecular weight of 82 kD, and one of the leucine rich glycoproteins (LRG). By reverse transcription-polymerase chain reaction (RT-PCR), GPV cDNA was amplified from mRNA of platelets and megakaryocytic cell lines. However, since there are few reports indicating whether GPV protein is expressed in megakaryocytes as a lineage and maturation specific protein, we studied the GPV expression at the protein level by using a novel monoclonal antibody (1D9) recognizing GPV. Flow cytometric and immunohistochemical analysis indicated that GPV was detected on the surface and in the cytoplasm of only the megakaryocytes in bone marrow aspirates. In a megakaryocytic cell line UT-7, GPV antigen increased after treatment with phorbol-12-myri-state-13-acetate (PMA). These data indicate that only megakaryocytes specifically express the GPV protein among hematopoietic cells and that the expression of GPV increases with differentiation of the megakaryocyte as GPIb-IX complex.

DNA sequence of the androgen receptor in prostatic tumor cell lines and tissue specimens assessed by means of the polymerase chain reaction

The Prostate ◽  
1993 ◽  
Vol 22 (1) ◽  
pp. 11-22 ◽  
Author(s):  
Zoran Culig ◽  
Helmut Klocker ◽  
Johannes Eberle ◽  
Felizia Kaspar ◽  
Alfred Hobisch ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Androgen Receptor ◽  
Tumor Cell ◽  
Dna Sequence ◽  
Cell Lines ◽  
Tumor Cell Lines ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Prostatic Tumor ◽  
Tissue Specimens

scholarly journals LncRNA PVT1 Promotes Hypoxia-Induced Cardiomyocyte Injury by Inhibiting miR-214-3p

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Chuanliang Liu ◽  
Jieqiong Zhang ◽  
Xuejie Lun ◽  
Lei Li
Keyword(s):  
Cell Viability ◽  
Cell Counting ◽  
Luciferase Reporter ◽  
H9c2 Cell ◽  
H9c2 Cells ◽  
Flow Cytometry Analysis ◽  
Chain Reaction ◽  
Cell Vitality ◽  
Gene Assay ◽  
Polymerase Chain

Objective. To explore the effect and related mechanism of LncRNA PVT1 on hypoxia-induced cardiomyocyte injury. Methods. PVT1RNA and miR-214-3p levels were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Cell vitality and apoptosis were, respectively, evaluated by Cell Counting Kit-8 (CCK-8) and flow cytometry analysis. Starbase and Dual luciferase reporter (DLR) gene assay was employed to validate the interaction between miR-214-3p and PVT1. Results. PVT1 was statistically upregulated, and miR-214-3p was statistically downregulated in hypoxia-induced H9c2 cells. The survival rate of H9c2 cells induced by hypoxia decreased statistically, while the apoptosis rate increased statistically ( P < 0.05 ). PVT1 knockdown upregulated the hypoxia-induced H9c2 cell viability and inhibited apoptosis. DLR assay verified the targeting relationship between PVT1 and miR-214-3p. In addition, miR-214-3p inhibitors reversed the viability of H9c2 cells with PVT1 knockout and promoted apoptosis. Conclusion. Silencing PVT1 can enhance the hypoxia-induced H9c2 cell viability and inhibit apoptosis, providing a potential target for the treatment of cardiovascular diseases.

scholarly journals Precision of genotyping of Epstein-Barr virus by polymerase chain reaction using three gene loci (EBNA-2, EBNA-3C, and EBER): predominance of type A virus associated with Hodgkin's disease [published erratum appears in Blood 1993 Oct 1;82(7):2268]

Blood ◽  
1993 ◽  
Vol 81 (12) ◽  
pp. 3372-3381 ◽  
Author(s):  
JC Lin ◽  
SC Lin ◽  
BK De ◽  
WC Chan ◽  
BL Evatt ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Cell Lines ◽  
Epstein Barr Virus ◽  
Point Mutations ◽  
Type A ◽  
Type B ◽  
Chain Reaction ◽  
Barr Virus ◽  
Polymerase Chain ◽  
Epstein Barr

Abstract To precisely determine the genotype of Epstein-Barr virus (EBV) in Hodgkin's disease (HD), we simultaneously analyzed three divergent gene loci (EBNA-2, EBNA-3C, and EBER) that distinguish type A and B viruses. The primers designed to amplify these three gene loci encompass either type-specific deletion sequences (EBNA-2 and EBNA-3C) or type-specific point mutations (EBER) that identify the virus strain based on the sizes of the polymerase chain reaction (PCR)-amplified products or the mobility shifts in single-strand conformation polymorphism analysis. The locations of point mutations were identified by direct sequencing of the PCR-amplified DNA. We analyzed 15 EBV-infected cell lines and found a good correlation between EBNA-2 and EBNA-3C typing results. In contrast, approximately 33% of the cell lines analyzed maintained type A sequences in EBNA-2 and EBNA-3C genes while carrying type B sequences in the EBER region. Data obtained from analysis of cell lines served as a reference for studying HD samples. EBV DNA was detected in about 70% of HD. Among the EBV-positive samples, 56% were associated with type A virus, 13% with type B, and 31% with dual viral sequences. Thus, type A virus is predominant in HD. Based on the histology, the frequencies of EBV positivity were 83%, 71%, and 33% for mixed cellularity, nodular sclerosis, and lymphocyte predominance, respectively. The detection of high frequency of both type A and B sequences in HD may provide a lead in investigating the role of dual viral infection in EBV pathogenesis.

Haptoglobin Genotypes in Nasopharyngeal Carcinoma

2009 ◽  
Vol 24 (1) ◽  
pp. 32-37 ◽  
Author(s):  
Ching-Chih Lee ◽  
Hon-Yi Lin ◽  
Shih-Kai Hung ◽  
Dian-Kun Li ◽  
Hsu-Chueh Ho ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Prognostic Factor ◽  
Nasopharyngeal Carcinoma ◽  
Primary Tumor ◽  
Tumor Volume ◽  
Prognostic Significance ◽  
Chain Reaction ◽  
Primary Tumor Volume ◽  
Negative Prognostic Factor ◽  
Polymerase Chain

Aim Haptoglobin polymorphisms are associated with different cancers; however, the occurrence of nasopharyngeal carcinoma (NPC) in relation to haptoglobin polymorphisms has not been reported. In this study, the distribution of haptoglobin genotypes among patients with NPC was investigated and the prognostic significance of haptoglobin genotypes was further analyzed. Material and methods Haptoglobin genotypes were analyzed using polymerase chain reaction and electrophoresis. The genotypes were determined in the sera of 49 NPC patients and in 134 controls. Results The haptoglobin genotypes of patients with NPC were as follows: Hp 1–1, 2%; Hp 2–1, 39%; Hp 2–2, 59%. The frequency of the Hp 2–2 genotype was much higher in NPC patients than in control individuals (p=0.044). Furthermore, NPC patients with the Hp 2–2 genotype had advanced T stages (p=0.001) and larger primary tumor volumes (p=0.035) than those with Hp 2–1 or 1–1. Conclusion An increased frequency of the Hp 2–2 genotype was associated with NPC. The Hp 2 allele was also overexpressed in NPC patients. NPC patients with the Hp 2–2 genotype had advanced T stage and a larger primary tumor volume. Hp 2–2 may be a negative prognostic factor in NPC.

scholarly journals Expression of the collagen receptor glycoprotein VI during megakaryocyte differentiation

Blood ◽  
2000 ◽  
Vol 96 (8) ◽  
pp. 2740-2745 ◽  
Author(s):  
Oscar Berlanga ◽  
Regis Bobe ◽  
Marion Becker ◽  
George Murphy ◽  
Mireille Leduc ◽  
...  
Keyword(s):  
Cell Lines ◽  
Chain Reaction ◽  
Activated T Cells ◽  
Glycoprotein Vi ◽  
Venom Toxin ◽  
Hel Cells ◽  
Snake Venom Toxin ◽  
Polymerase Chain ◽  
Collagen Receptor

Abstract This study examined the expression of the platelet collagen receptor glycoprotein VI (GPVI) in megakaryocyte cell lines and primary megakaryocytes by reverse transcriptase-polymerase chain reaction and by flow cytometry and ligand blotting using the snake venom toxin convulxin. Expression of GPVI is increased in the megakaryoblastic cell lines HEL and CMK on differentiation with the phorbol ester phorbol 12-myristate 13-acetate (PMA), along with the Fc receptor γ-chain (FcR γ-chain). The increase in GPVI expression is associated with marked potentiation of tyrosine phosphorylation and Ca++ elevation in response to convulxin. Syk, linker for activated T cells, and phospholipase Cγ2 (PLCγ2) are among the proteins tyrosine phosphorylated on convulxin stimulation in PMA-differentiated HEL cells. Studies on primary murine megakaryocytes grown in vitro confirmed that GPVI is up-regulated in parallel with functional activation, assessed by measurement of [Ca++]i, during differentiation. The results demonstrate that expression of GPVI is up-regulated along with the FcR γ-chain during differentiation of megakaryocytes.

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