H. pylori-induced apoptosis in human gastric cancer cells mediated via the release of apoptosis-inducing factor from mitochondria.
OBJECTIVE
Our previous study of Helicobacter pylori-induced apoptosis showed the involvement of Bcl-2 family proteins and cytochrome c release from mitochondria. Here, we examine the release of other factors from mitochondria, such as apoptosis-inducing factor (AIF), and upstream events involving caspase-8 and Bid.


METHODS
Human gastric adenocarcinoma (AGS) cells were incubated with a cagA-positive H. pylori strain for 0, 3, 6, and 24 hours and either total protein or cytoplasmic, nuclear, and mitochondrial membrane fractions were collected.


RESULTS
Proteins were immunoblotted for AIF, Bid, polyadenosine ribose polymerase (PARP), caspase-8, and beta-catenin. H. pylori activated caspase-8, caused PARP cleavage, and attenuated mitochondrial membrane potential. A time-dependent decrease in beta-catenin protein expression was detected in cytoplasmic and nuclear extracts, coupled with a decrease in beta-actin. An increase in the cytoplasmic pool of AIF was seen as early as 3 hours after H. pylori exposure, and a concomitant increase was seen in nuclear AIF levels up to 6 hours. A band corresponding to full-length Bid was seen in both the cytoplasmic and the nuclear fractions of controls, but not after H. pylori exposure. Active AIF staining was markedly increased in gastric mucosa from infected persons, compared to uninfected controls.


CONCLUSIONS
H. pylori might trigger apoptosis in AGS cells via interaction with death receptors in the plasma membrane, leading to the cleavage of procaspase-8, release of cytochrome c and AIF from mitochondria, and activation of subsequent downstream apoptotic events, as reported previously for chlorophyllin. This is consistent with AIF activation that was found in the gastric mucosa of humans infected with H. pylori. Hence, the balance between apoptosis and proliferation in these cells may be altered in response to injury caused by H. pylori infection, leading to an increased risk of cancer.
Ashktorab
Hassan
H
Cancer Center and Gastrointestinal Division, Department of Medicine, Howard University, Washington, DC 20060, USA. hashktorab@howard.edu
Dashwood
Rod H
RH
Dashwood
Mohaiza M
MM
Zaidi
Syed I
SI
Hewitt
Stephen M
SM
Green
William R
WR
Lee
Edward L
EL
Daremipouran
Mohammadreza
M
Nouraie
Mehdi
M
Malekzadeh
Reza
R
Smoot
Duane T
DT
D017209:Apoptosis; D051033:Apoptosis Inducing Factor / Q000378:metabolism*; D002458:Cell Fractionation; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000737:chemistry; D003593:Cytoplasm / Q000737:chemistry; D004847:Epithelial Cells / Q000382:microbiology*; D016480:Helicobacter pylori / Q000502:physiology*; D006801:Humans; D015151:Immunoblotting; D008928:Mitochondria / Q000378:metabolism*; D009685:Nuclear Envelope / Q000737:chemistry
D051033:Apoptosis Inducing Factor