Influence of tissue preparation techniques on p53 expression in bronchial and bladder carcinomas, assessed by immunofluorescence staining and flow cytometry

Cytometry ◽  
1991 ◽  
Vol 12 (7) ◽  
pp. 622-627 ◽  
Author(s):  
Odd Mørkve ◽  
Jens Høstmark
Keyword(s):  
Flow Cytometry ◽  
Immunofluorescence Staining ◽  
Tissue Preparation ◽  
P53 Expression ◽  
Bladder Carcinomas ◽  
Preparation Techniques

Comparison of Various Tissue-Preparation Techniques for Cryosectioning of Frozen Mouse Tissues

2009 ◽  
Vol 32 (4) ◽  
pp. 186-189 ◽  
Author(s):  
Lee Hui Ling ◽  
Susan Mary Rogers ◽  
Vanessa Tay ◽  
Gino V. Limmon ◽  
Zheng Dahai ◽  
...  
Keyword(s):  
Tissue Preparation ◽  
Mouse Tissues ◽  
Preparation Techniques

scholarly journals CXCL12-CXCR4-Mediated Chemotaxis Supports Accumulation of Mucosal-Associated Invariant T Cells Into the Liver of Patients With PBC

2021 ◽  
Vol 12 ◽  
Author(s):  
Zhilei Chen ◽  
Suying Liu ◽  
Chengmei He ◽  
Jinlei Sun ◽  
Li Wang ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
Cytokine Production ◽  
Immunofluorescence Staining ◽  
Primary Biliary Cholangitis ◽  
Hepatic Hemangioma ◽  
Cxcr4 Antagonist ◽  
Mait Cells ◽  
Ifn Γ ◽  
Invariant T Cells

Objectives: To explore the potential role of CD3+CD8+CD161high TCRVα7.2+ mucosal-associated invariant T (MAIT) cells in the pathogenesis of primary biliary cholangitis (PBC).Methods: We enrolled 55 patients with PBC, 69 healthy controls (HCs), and 8 patients with hepatic hemangioma. Circulating MAIT cells and their chemokine receptor profiles and cytokine production were quantified using flow cytometry. Liver-resident MAIT cells were examined by immunofluorescence staining. CXCL12-mediated chemotaxis of MAIT cells was measured using a transwell migration assay. Plasma interleukin (IL)-18 was measured using ELISA, and cytokine production in IL-18-stimulated MAIT cells was detected using flow cytometry.Result: Peripheral MAIT cells were found to be significantly lower in patients with PBC (3.0 ± 3.2% vs. 9.4 ± 8.0%, p < 0.01) and negatively correlated with alkaline phosphatase (ALP) levels (r = −0.3209, p < 0.05). Liver immunofluorescence staining suggested that MAIT cells might accumulate in PBC liver. MAIT cells from patients with PBC expressed higher levels of CXCR4 (84.8 ± 18.0% vs. 58.7 ± 11.4%, p < 0.01), and the expression of CXCL12 was higher in PBC liver. CXCL12 promoted MAIT cell chemotaxis (70.4 ± 6.8% vs. 52.2 ± 3.5%, p < 0.01), which was attenuated by CXCR4 antagonist. MAIT cells from PBC produced significantly more interferon-γ (IFN-γ) (88.3 ± 4.2% vs. 64.2 ± 10.1%, p < 0.01), tumor necrosis factor-α (TNF-α) (93.0 ± 1.1% vs. 80.1 ± 5.3%, p < 0.01), Granzyme B (89.3 ± 3.3% vs. 72.1 ± 7.0%, p < 0.01), and perforin (46.8 ± 6.6% vs. 34.8 ± 7.7%, p < 0.05). MAIT cells from PBC expressed higher levels of IL18-Rα (83.8 ± 10.2% vs. 58.3 ± 8.7%, p < 0.01). Plasma IL-18 was more abundant in patients with PBC (286.8 ± 75.7 pg/ml vs. 132.9 ± 78.1 pg/ml, p < 0.01). IL-18 promoted IFN-γ production in MAIT cells (74.9 ± 6.6% vs. 54.7 ± 6.7%, p < 0.01), which was partially attenuated by blocking IL-18R (68.6 ± 8.3% vs. 43.5 ± 4.2%, p < 0.01).Conclusion: Mucosal-associated invariant T cells from patients with PBC accumulated in the liver via CXCL12-CXCR4-mediated chemotaxis, produced pro-inflammatory cytokines, and contributed to portal inflammation, which was potentially mediated by elevated IL-18. Targeting MAIT cells might be a therapeutic approach for PBC.

scholarly journals Endoplasmic Reticulum Stress Is Involved in Glucocorticoid-Induced Apoptosis in PC12 Cells

2021 ◽  
Vol 2021 ◽  
pp. 1-10
Author(s):  
Shanyong Yi ◽  
Weibo Shi ◽  
Min Zuo ◽  
Songjun Wang ◽  
Rufei Ma ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Endoplasmic Reticulum ◽  
Endoplasmic Reticulum Stress ◽  
Pc12 Cells ◽  
Neuronal Injury ◽  
Immunofluorescence Staining ◽  
High Concentration ◽  
Flow Cytometry Assay ◽  
Apoptosis Rate ◽  
Amino Terminal

Objective. The present study selected PC12 cells to construct a neuronal injury model induced by glucocorticoids (GC) in vitro, aiming to explore whether the endoplasmic reticulum stress (ERS) PKR-like endoplasmic reticulum kinase (PERK)-activating transcription factor 4 (ATF4)-C/EBP-homologous protein (CHOP) and inositol requirement 1 (IRE1)-apoptosis signal regulating kinase 1 (ASK1)-C-Jun amino-terminal kinase (JNK) signaling pathways are associated with the neuronal injury process induced by GC and provide morphological evidence. Methods. Cell models with different doses and different durations of GC exposure were established. The viability of PC12 cells was detected by the CCK-8 assay, and the apoptosis rate of PC12 cells was detected by the flow cytometry assay. The expression of microtubule-associated protein 2 (Map2); glucocorticoids receptor (GR); cellular oncogene fos (C-fos); and ERS-related proteins, glucose-regulated protein 78 (GRP78), p-PERK, p-IRE1, ATF4, ASK1, JNK, and CHOP, was observed by immunofluorescence staining. Results. The results of immunofluorescence staining showed that PC12 cells abundantly expressed Map2 and GR. The CCK-8 assay revealed that high-concentration GC exposure significantly inhibited the cell viability of PC12 cells. The flow cytometry assay indicated that high-concentration GC exposure significantly increased the apoptosis rate of PC12 cells. Immunofluorescence staining showed that GC exposure significantly increased the expression of C-fos, GRP78, p-PERK, p-IRE1, ATF4, ASK1, JNK, and CHOP. Treatment with ERS inhibitor 4-phenylbutyric acid (4-PBA) and GR inhibitor RU38486 attenuated related damage and downregulated the expression of the abovementioned proteins. Conclusion. High-concentration GC exposure can significantly inhibit the viability of PC12 cells and induce apoptosis. PERK-ATF4-CHOP and IRE1-ASK1-JNK pathways are involved in the above damage process.

Flow cytometry, cellular DNA content, and prognosis in human malignancy.

1987 ◽  
Vol 5 (10) ◽  
pp. 1690-1703 ◽  
Author(s):  
D E Merkel ◽  
L G Dressler ◽  
W L McGuire
Keyword(s):  
Flow Cytometry ◽  
Data Base ◽  
Dna Content ◽  
Dna Analysis ◽  
Clinical Decision Making ◽  
Risk Groups ◽  
Clinical Decision ◽  
Tumor Type ◽  
Cellular Dna ◽  
Bladder Carcinomas

The use of flow cytometry to analyze the cellular DNA content of human malignancies has become increasingly commonplace. The relationship between abnormalities in DNA content or proliferative characteristics and prognosis is becoming clear for a variety of malignancies in part through new techniques that permit analysis of archival material. High- and low-risk groups of patients with early breast and bladder carcinomas, non-small-cell lung cancer, and colorectal, ovarian, and cervical carcinoma can be distinguished on the basis of abnormal stemline DNA content. In several hematologic and common pediatric malignancies, the prognostic relevance of DNA content flow cytometry has been similarly established. Though the interpretation of tumor cell cycle analyses is less certain, this characteristic may also be prognostically important. However, generalizations cannot be made when applying flow cytometric DNA analysis to clinical decision making. The prognostic importance of an abnormal DNA histogram for an individual patient must be assessed on the basis of the relevant data base for that particular tumor type. The current extent of this data base for various malignancies is reviewed.

Study of Cellular DIMA Content by Flow Cytometry in Primary Bladder Carcinomas

1992 ◽  
Vol 21 (1) ◽  
pp. 58-63 ◽  
Author(s):  
R. Campanella ◽  
A. Russo ◽  
S. Plaja ◽  
V. Bazan ◽  
C. Pavone ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Primary Bladder ◽  
Bladder Carcinomas

Prognostic Implications of Aberrations in p16/pRb Pathway in Urothelial Bladder Carcinomas: A Multivariate Analysis Including p53 Expression and Proliferation Markers

2001 ◽  
Vol 39 (2) ◽  
pp. 167-177 ◽  
Author(s):  
Penelope Korkolopoulou ◽  
Panayota Christodoulou ◽  
Andreas Lazaris ◽  
Euphemia Thomas-Tsagli ◽  
Panayotis Kapralos ◽  
...  
Keyword(s):  
Multivariate Analysis ◽  
Proliferation Markers ◽  
P53 Expression ◽  
Urothelial Bladder ◽  
Prognostic Implications ◽  
Bladder Carcinomas

p53 Expression in Epithelial Ovarian Neoplasms: Relationship to Clinical and Pathological Parameters, Ki-67 Expression and Flow Cytometry

1994 ◽  
Vol 53 (3) ◽  
pp. 301-306 ◽  
Author(s):  
Rudi Henriksen ◽  
Peter Strang ◽  
Erik Wilander ◽  
Torbjörn Bäckström ◽  
Bernhard Tribukait ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Ovarian Neoplasms ◽  
Ki 67 ◽  
P53 Expression

scholarly journals Hypericin Exerts Detrimental Effect on Huh-7 As a Delegacy of Hepatocellular Carcinoma: A P53 Dependent Pathway

2020 ◽  
Vol 9 ◽  
pp. 1896
Author(s):  
Maedeh Olya ◽  
Hamid Zaferani Arani ◽  
Amirhossein Shekarriz ◽  
Amirhossein Zabolian ◽  
Hadi Zare Marzouni ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Flow Cytometry ◽  
Liver Cancer ◽  
Cell Line ◽  
Real Time ◽  
Real Time Pcr ◽  
Huh7 Cell ◽  
P53 Expression ◽  
Huh7 Cells ◽  
Huh7 Cell Line

Background: Hepatocellular carcinoma is the most common type of liver cancer which arises from the main cells in the liver. We address many studies investigating anti-cancer role of hypericin, however the proposing corresponding molecular pathway seems to be still a debate. Therefore, the present study aimed to evaluate the apoptotic effect of hypericin on the Huh7 as the liver cancer cell line and its relation with the gate keeper gene P53. Materials and Methods: In this study, the Huh7 cell line and fibroblast cells (as control group) were treated with different concentrations of hypericin for 24 and 48 hours. Detection of cell death was performed by MTT assay and flow cytometry. The expression of bax, bcl2 and p53 mRNAs was evaluated by Real-time PCR. Also, Immunocytochemistry (ICC) analysis was used for further evaluation of P53expression. Results: The results showed that hypericin has a dose-dependent cytotoxic effect on the Huh7 cell line, with no or marginal effect on fibroblastic cells. According to flow cytometry results, about 53%cells underwent apoptosis after exposure to LD50 of hypericin for 24 hours. Real-time PCR data demonstrated that the pro-apoptotic genes Bax and P53 expression level increased. Expectedly ICC results confirmed the up-regulation of P53 proteins in treated samples. Conclusion: Our results indicate the cytotoxicity of hypericin on Huh7 cells by affecting the expression of the gate keeper gene P53; furthermore it is suggested that this herb can be utilized simultaneously with modalities targeting P53 up-regulation or related molecular pathways. [GMJ.2020;9:e1896]

Indici di proliferazione (Ki 67, TLI) ed espressione della p53. Esperienza personale: Proliferation indices (Ki 67, TLI) and p53 expression. Personal experience

1995 ◽  
Vol 62 (2) ◽  
pp. 234-239 ◽  
Author(s):  
B. Monica ◽  
R. Minari
Keyword(s):  
Flow Cytometry ◽  
Dna Content ◽  
Personal Experience ◽  
S Phase ◽  
Urothelial Bladder Cancer ◽  
Ki 67 ◽  
P53 Overexpression ◽  
P53 Expression ◽  
Proliferation Indices ◽  
Urothelial Bladder

The Authors report their experience on the study of urothelial bladder cancer by determining the overexpression of p53, DNA-content by flow-cytometry, Ki 67, TLI and cytofluorimetric S-phase in 81 patients. Ki 67, DNA content and p53 overexpression are statistically related with histologic grade, unlike TLI and S-phase. TLI, overexpression of p53 and Ki 67 are statistically related with stage, whereas ploidy and S-phase do not show any relation with stage. The Authors discuss these findings.

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