Detection of Pyroptosis by Measuring Released Lactate Dehydrogenase Activity

Rayamajhi, Manira; Zhang, Yue; Miao, Edward A.

doi:10.1007/978-1-62703-523-1_7

Cited by 144 publications

(111 citation statements)

References 9 publications

(10 reference statements)

Supporting

Mentioning

101

Contrasting

Unclassified

Order By: Relevance

“…Induction of pyroptosis is characterized by activation of caspase-1, loss of integrity of cell membrane, and release of lactate dehydrogenase (LDH) which is normally maintained within the cell cytosol. Thus the release of intracellular LDH is recognized as an effective methodology to define the occurrence of pyroptosis [14], which was performed in this study to define whether pyroptosis was induced. Our data showed that pyroptosis in these AIM2 overexpressed cells was induced in a time-dependent manner (Figure 3I).…”

Section: Resultsmentioning

confidence: 99%

Loss of AIM2 expression promotes hepatocarcinoma progression through activation of mTOR-S6K1 pathway

et al. 2016

View full text Add to dashboard Cite

Absent in melanoma (AIM2) is a member of the interferon-inducible HIN-200 protein family and is recently recognized to play an important dual role in both innate immunity and tumor pathology. However, the role of AIM2 in the development of hepatocellular carcinoma (HCC) remains to be clarified. Here we showed that AIM2 expression was significantly decreased in liver cancer tissues, and loss of its expression was significantly correlated with more advanced tumor progression. Exogenous overexpression of AIM2 in HCC cells suppressed mammalian target of rapamycin (mTOR)-S6K1 pathway and further inhibited proliferation, colony formation and invasion of HCC cells. On the contrary, block of AIM2 in HCC cells induced (mTOR)-S6K1 pathway activation and thus promoted HCC progression. Treatment with mTOR pathway inhibitor rapamycin further verified its contribution to HCC progression in AIM2 absent HCC cells. Thus, these data suggested that AIM2 played a critical role as a tumor suppressor and might serve as a potential therapeutic target for future development of AIM2-based gene therapy for human liver cancer. This study also paves a new avenue to treat AIM2-deficient cancer by suppression of mTOR.

show abstract

Section: Resultsmentioning

confidence: 99%

Loss of AIM2 expression promotes hepatocarcinoma progression through activation of mTOR-S6K1 pathway

et al. 2016

View full text Add to dashboard Cite

show abstract

“…One of the main morphological features of pyroptosis is loss of the integrity of the plasma membrane, which leads to cell lysis and the release of cellular contents to the surroundings1644. Lactate dehydrogenase (LDH) is an enzyme that is normally sequestered inside the cell and is released extracellularly only in the event of cell lysis45. Measuring the activity of LDH in the medium of the cultured cells is a useful tool for detecting lytic cell death, including pyroptosis45.…”

Section: Resultsmentioning

confidence: 99%

Hepatitis C Virus Infection of Cultured Human Hepatoma Cells Causes Apoptosis and Pyroptosis in Both Infected and Bystander Cells

Kofahi

Taylor

Hirasawa

et al. 2016

Sci Rep

View full text Add to dashboard Cite

Individuals infected with hepatitis C virus (HCV) are at high risk of developing progressive liver disease, including cirrhosis and hepatocellular carcinoma (HCC). How HCV infection causes liver destruction has been of significant interest for many years, and apoptosis has been proposed as one operative mechanism. In this study, we employed a tissue culture-adapted strain of HCV (JFH1T) to test effects of HCV infection on induction of programmed cell death (PCD) in Huh-7.5 cells. We found that HCV infection reduced the proliferation rate and induced caspase-3-mediated apoptosis in the infected cell population. However, in addition to apoptosis, we also observed infected cells undergoing caspase-1-mediated pyroptosis, which was induced by NLRP3 inflammasome activation. By co-culturing HCV-infected Huh-7.5 cells with an HCV-non-permissive cell line, we also demonstrated induction of both apoptosis and pyroptosis in uninfected cells. Bystander apoptosis, but not bystander pyroptosis, required cell-cell contact between infected and bystander cells. In summary, these findings provide new information on mechanisms of cell death in response to HCV infection. The observation that both apoptosis and pyroptosis can be induced in bystander cells extends our understanding of HCV-induced pathogenesis in the liver.

show abstract

“…Supernatant samples were collected at the indicated time points in figures. Cytotoxicity was defined as the percentage of total lactate dehydrogenase released into the supernatant and was determined as described (Rayamajhi et al, 2013b). IL-1β secretion was determined by enzyme-linked immunosorbent assay (ELISA) (R&D Systems).…”

Section: Methodsmentioning

confidence: 99%

Canonical Inflammasomes Drive IFN-γ to Prime Caspase-11 in Defense against a Cytosol-Invasive Bacterium

Aachoui

Kajiwara

Leaf

et al. 2015

Cell Host & Microbe

105

147

View full text Add to dashboard Cite

SUMMARY The inflammatory caspases-1 and -11 are activated in response to different agonists and act independently to induce pyroptosis. In the context of IL-1β/IL-18 secretion however, in vitro studies indicate that caspase-11 acts upstream of NLRP3 and caspase-1. By contrast, studying infection in vivo by the cytosol-invasive bacterium Burkholderia thailandensis, we find that caspase-1 activity is required upstream of caspase-11 to control infection. Caspase-1 activated IL-18 induces IFN-γ to prime caspase-11 and rapidly clear B. thailandensis infection. In the absence of IL-18, bacterial burdens persist, eventually triggering other signals that induce IFN-γ. Whereas IFN-γ was essential, endogenous type I interferons were insufficient to prime caspase-11. Although mice transgenic for caspase-4, the human orthologue of caspase-11, cleared B. thailandensis in vivo, they did not strictly require IFN-γ priming. Thus, caspase-1 provides priming signals upstream of caspase-11 but not caspase-4 during murine defense against a cytosol-invasive bacterium.

show abstract

Detection of Pyroptosis by Measuring Released Lactate Dehydrogenase Activity

Cited by 144 publications

References 9 publications

Loss of AIM2 expression promotes hepatocarcinoma progression through activation of mTOR-S6K1 pathway

Loss of AIM2 expression promotes hepatocarcinoma progression through activation of mTOR-S6K1 pathway

Hepatitis C Virus Infection of Cultured Human Hepatoma Cells Causes Apoptosis and Pyroptosis in Both Infected and Bystander Cells

Canonical Inflammasomes Drive IFN-γ to Prime Caspase-11 in Defense against a Cytosol-Invasive Bacterium

Contact Info

Product

Resources

About