Bcl-10 Mediates Aberrant NF- B Activity in Hepatocellular Carcinomas Nathan Wilds Summer 2004.
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Transcript of Bcl-10 Mediates Aberrant NF- B Activity in Hepatocellular Carcinomas Nathan Wilds Summer 2004.
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Bcl-10 Mediates Aberrant NF-Bcl-10 Mediates Aberrant NF-B B Activity in Hepatocellular Activity in Hepatocellular
CarcinomasCarcinomas
Nathan Wilds
Summer 2004
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Carcinomas of the liver and hepatic bile duct are 5th most common GI malignancies and 3rd most lethal neoplasm of GI tract.
In the midsouth an increase in HCCs is anticipated due to an increase in Hepatitis B and C infections.
NF-B is directly involved in normal immune and inflammatory responses, cell proliferation, differentiation, apoptosis, and oncogenesis.
NF-B is constitutively activated in murine and human HCCs.
IntroductionIntroduction
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Identify crucial upstream and downstream components of the NF-B Pathway in murine
HCCs derived from bitransgenic animals. These animals are a widely used model of
liver carcinogenesis.
What is the mechanism of NF-B constitutive activation in cancer cells???
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IKK complex
IB
NF-B
IBpp
IB UbUb
UbUb
Ub
UbUb
UbUb
Ub
pp
IB UbUb
UbUb
Ub
UbUb
UbUb
Ub
pp
NF-B
NF-B
NF-B
Degradation(26S proteosome)
Nuclear translocation
Canonical pathway of NF-kB Activation
Cytosol
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Gene Array Shows Gene Expression Increasing Gene Array Shows Gene Expression Increasing From Surrounding Liver to Tumor.From Surrounding Liver to Tumor.
Transforming growth factor α Transforming growth factor β1 Cytochrome P450 (several variances) B-cell leukemialymphoma 10 (Bcl-10) Myeloid cell leukemia sequences Glucokinase (Gck) Deoxyribonuclease II alpha (DNase2a) Procollagen, type IV, 1 Carboxylesterase 3 (Ces3) Glutathione transferase
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Bcl-10IKK complex
IB
NF-B
IBpp
IB UbUb
UbUb
Ub
UbUb
UbUb
Ub
pp
IB UbUb
UbUb
Ub
UbUb
UbUb
Ub
pp
NF-B
UbUb
UbUb
UbUb
Ub
NF-B
NF-B
Degradation(26S proteosome)
Nuclear translocation
Role of Bcl-10 in the NF-B pathway.
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K562 WEHI t1 t2 t3 t4 t5 ma3 ma2 ma1 wt3 wt2 wt1
K562 WEHI t6 t7 t8 t4 t5 ma3 ma2 ma1 wt3 wt2 wt1
Bcl-10 is Over Expressed in Tumors When Compared to Dysplastic or Wild-Type Livers.
IB: Bcl-10
IB: Bcl-10
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SL T SL T
Bcl-10 Expression in Bitransgenic Mice Comparing Surrounding Liver and Tumor Samples
IB Bcl-10
PCR Bcl-10
SL T SL T SL T SL T
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IP IKK-γ + + +
223 T SL
IKK-γ
IB: Ubiquitan
IKK-γ is Ubiquitinated in Tumor, but Not in Surrounding Liver.
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0%
25%
50%
75%
100%
125%
% L
uc
ife
ras
e A
cti
vit
y r
ela
tiv
e t
o c
on
tro
l
none
5ng C IKK
50ng C IKK
Regulation of Regulation of FF--B ActivityB Activity
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ConclusionsConclusions
Bcl-10 is overexpressed in tumors relative to surrounding livers as well as dysplastic and wt tumors,
Bcl10 is involved in the uibiquitination of IKK- in tumor samples,
IKK regulates NF-B activity in HCCs. Future experiments will determine whether Bcl-10
is involved in NF-B signaling in malignant hepatocytes. email
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Acknowledgements Acknowledgements
The University of Tennessee Health Science Center, Dept of Pharmacology
ASPET Dr. Marcello Arsura and all the post-docs in his
laboratory; Drs. Swayamjot Kaur, Venkatramen Manickam, and Fang Wang, for all their answers to my “does this look right?” and “what in the world is this?” questions.
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Questions???Questions???