Is autophagy caspase dependent?
Dysregulated autophagy contributes to caspase-dependent neuronal apoptosis. Cell Death Dis. 2018 Dec 11;9(12):1189. doi: 10.1038/s41419-018-1229-y.
How is autophagy measured?
One of the most common ways to monitor autophagy is by measuring the protein levels of LC3-II, which is incorporated into autophagosomes and then degraded in the lysosome. There are many different ways to measure LC3 protein levels, including western blot and immunofluorescent microscopy2.
What is p62 in autophagy?
p62 is a receptor for cargo destined to be degraded by autophagy, including ubiquitinated protein aggregates destined for clearance. The p62 protein is able to bind ubiquitin and also to LC3, thereby targeting the autophagosome and facilitating clearance of ubiquitinated proteins.
What do caspases do?
Caspases (cysteine-aspartic proteases) are proteolytic enzymes largely known for their role in controlling cell death and inflammation. Based on their function, mammalian caspase-2, -3, -7, -8, -9 and -10 are apoptotic caspases, where as caspase-1, -4, -5, -11 and -12 are involved in inflammation.
How do you know if you are in autophagy?
Depending on the individual’s metabolism, significant autophagy may take two to four days of fasting in humans. Autophagy is believed to begin when glucose and insulin levels drop considerably. Animal studies have shown evidence of autophagy after 24 hours of fasting, which starts peaking at around 48 hours of fasting.
How many autophagosomes are in a cell?
Table 1.
| Variable | Unit | Induced |
|---|---|---|
| Autophagosome flux, J | A/h/cell | 105.4 |
| Number of autophagosomes, nA | A/cell | 16.7 |
| Number of autolysosomes, nAL | AL/cell | 251.2 |
| Average volume of an autophagosome | fL | 4.7 |
How is ATG7 related to autophagy?
Autophagy related 7 is a protein in humans encoded by ATG7 gene. During the initiation of autophagy, ATG7 acts like an E-1 enzyme for ubiquitin-like proteins (UBL) such as ATG12 and ATG8. ATG7 helps these UBL proteins in targeting their molecule by binding to them and activating their transfer to an E-2 enzyme.
Does p62 increase with autophagy?
This experiment was conducted under reducing conditions. An advantage to monitoring p62 to measure autophagic flux is that lysosomal inhibitors are not necessary, because unlike LC3-II, p62 does not usually increase when autophagy is induced.
Does p62 increase during autophagy?
Since p62 is itself an autophagy substrate, impaired autophagy is associated with increased levels of p62 protein in mammals and Drosophila (Klionsky et al., 2016). Indeed, we observed that flight muscles from middle-aged Drosophila show elevated dP62 levels (Figures 2A′–2C′; quantification in Figures S2B and S2C).
What is the role of autophagy in cancer cells?
Autophagy1 is a complex of adaptive cellular response that enhances cancer cell survival in the face of cellular stresses such as chemothery. Here we show that in human gallbladder carcinoma (GBC) cells lines, SGC-996 and GBC-SD, autophagy is induced by the DNA damaging agent 5-fluorouracil (5-FU).
How is Ras related to autophagic flux in cancer?
Oncogenic RAS has previously been shown to regulate autophagic flux, and cancers with high incidence of RAS mutations, such as pancreatic cancer, have been described in the literature as being particularly susceptible to CQ treatment, leading to the hypothesis that oncogenic RAS makes cancer cells dependent on autophagy.
Which is the agent that induces autophagy in GBC cells?
Here we show that in human gallbladder carcinoma (GBC) cells lines, SGC-996 and GBC-SD, autophagy is induced by the DNA damaging agent 5-fluorouracil (5-FU). While in combination with the pre-treatment of chloroquine (CQ), a inhibitor of autophagy, the inhibition of 5-FU to the proliferation and viability of GBC cells was potentiated.
What are the abbreviations for autophagy and chloroquine sensitivity?
Abbreviations: ATG autophagy-related BafA1 bafilomycin A1 CQ chloroquine HCQ hydroxychloroquine HEK immortalized embryonic kidney cells