POSTERS

P589

A MULTISITE STUDY OF NON-CIRRHOTIC HEPATOCELLULAR

CARCINOMA (HCC) IN PATIENTS WITH AND WITHOUT

TRADITIONAL RISK FACTORS

E. Connor1, T. Peeraphatdit2, A. Chan2, K. Janssen2, J. Albrecht3,

G. Punjabi3, C. Smith2, P. Ricci4. 1Gastroenterology, 2University

of Minnesota, 3Hennepin County Medical Center, 4Veterans Affairs

Medical Center Minneapolis, Minneapolis, MN, United States

E-mail: estherekao@gmail.com

Background and Aims: Non-cirrhotic HCC develops with and

without traditional risk factors (TRF, ie. HBV, HCV, alcohol and

hemochromatosis). We recently conducted a retrospective study

of non-cirrhotic HCC (Am. Coll. Gastro. 2013 abstract 477) at

our VAMC. The current study includes two additional centers in

Minneapolis: HCMC and UMMC.

Aims:

1. Determine prevalence of non-cirrhotic HCC in a larger

population.

2. Compare natural history and metabolic syndrome (MS)

prevalence in non-cirrhotic HCC subjects with and without TRF.

Methods: Multisite retrospective review of subjects diagnosed with

HCC (ICD-9 155.0) from 2000–2012. Subjects without biochemical,

clinical, radiological evidence of cirrhosis were included in the

study and divided into two groups: subjects with TRF and subjects

without TRF.

Results: Of 1,088 HCC total subjects, 153 (14%) were non cirrhotic.

Of these, 59% had TRF (HCV 58%, HBV 23%, alcohol 51%) and 41%

were without TRF. Subjects without TRF were older, had larger

tumors (9.1 cm vs. 6 cm, p =0.021), more dyslipidemia (62% vs.

27%, p = 0.000) and an increased trend for diabetes (46% vs. 31%,

p = 0.076). Non-cirrhotic subjects with and without TRF received

curative treatments in similar proportions (25 vs. 20%) and there

was no survival difference (8.5 vs.11 months, p = 0.688).

Conclusions: In a larger and more diverse population, the

prevalence of non-cirrhotic HCC and HCC without TRF was similar

to our previously presented data. Subjects without TRF did not have

a survival advantage. Large tumor size, advanced age and possibly

metabolic syndrome may affect survival of non-cirrhotic HCC

without TRF; a prospective study is needed to test this hypothesis.

P590

RELATION OF HCV INDUCED INSULIN RESISTANCE AND

HEPATOCELLULAR CARCINOMA: ROLE OF MDR1 GENE c.335T>C

AND c.3073A>C SNPS

A.Z. Elsamanoudy1, A.A. Atwa2, H. Elalfy2, A.H.A. Metwali2. 1Medical

Biochemistry, 2Mansoura University, Mansoura, Egypt

E-mail: elalfy_hatem66@yahoo.com

Background and Aims: Hepatocellular carcinoma (HCC), is the

most common primary liver tumor. HCV-associated insulin

resistance (IR) may cause hepatic steatosis, hepatic fibrosis and

hepatocarcinogenesis. The multidrug resistance 1 gene (MDR1) is a

candidate gene for susceptibility to HCC.

The aim of the current study was to evaluate the association of the

MDR 1 gene c.335T>C and the c.3073A>C SNPs with HCV induced

HCC and to correlate this to insulin resistance state.

Methods: A total of 205 HCC patients (on top of HCV) were enrolled

in this study. Genotyping of MDR1 gene SNPs was done by PCR-

RFLP.

Results: Results revealed that there was a significantly increased

risk of HCC in chronic HCV with hepatic steatosis. The CC genotype

of the c.335T>C polymorphism was associated with an increased

risk of developing HCC compared to the TT genotype.

Conclusions: HCV-related metabolic complications as hepatic

steatosis and IR may be associated with increased risk of HCC

development. c.335T>C and c.3073A>C SNPs of MDR1 gene could

be considered as a possible molecular candidates for the HCC

development in chronic HCV patients.

P591

IMPROVEMENT OF TRANSDUCTION WITH Ad-GFP VECTOR

MEDIATED BY siRNA- IFN-a IN HEPATIC CELLS

A.A. Sobrevilla Navarro, A.S. Sandoval Rodriguez, L.D. Hernandez

Ortega, J. Armendariz Borunda, A.M. Salazar Montes. University of

Guadalajara, Guadalajara, Mexico

E-mail: asalazar_montes@hotmail.com

Background and Aims: Adenoviruses (Ad) are the most common

vectors used in clinical trials for gene therapy. Ad have shown

to have high tropism for liver being the ideal vector to delivery

therapeutic genes to this organ. Interferon type 1 (a and b) play

an important role in the elimination of adenovirus by the immune

response of the organism.

Methods: Huh7 cells were cultured in DMEM, 5% FBS at 37°C

and 5% CO2, transfected with 70nM of IFNa or irrelevant siRNA,

incubated for six hours and then exposed to 1×109 vp/ml of rAd-

GFP for 24 hrs. Expression of IFNa1 and TNF-a were determined

by qRT-PCR. Cell transduction was analyzed by flow cytometry (FC)

and qPCR and GFP protein by western blot.

Results: 70 nM of IFNa1-siRNA inhibited 96% of IFNa1 gene

expression (p < 0.001) and 65% of TNF-a (p < 0.05) compared to

irrelevant-siRNA. Ad-GFP transduction measured by FC and q-PCR

increased 39.2% and 27% respectively in IFNa1-siRNA treatment

compared to control. GFP protein increased 50% when IFNa1-siRNA

was used compared to control.

Conclusions: Inhibition of IFNa mRNA with IFNa1-siRNA permits

a higher transgene expression (GFP) indicating the crucial role of

IFNa on adenovirus elimination in transduced cells. This strategy

could be useful in clinical trials conducted for liver diseases, where

adenovirus are used as vector for therapeutic genes; allowing an

increased transgene expression leading to better results in the

resolution liver diseases.

4B. MOLECULAR AND CELLULARBIOLOGY: HSCS AND FIBROSIS

P592

3,3′-DIINDOLYLMETHANE MODULATES THE IMBALANCE OF

TREGS TO Th17 CELLS DURING LIVER FIBROGENESIS THROUGH

ACTIVATING THE ARYL HYDROCARBON RECEPTOR AND

INHIBITING TOLL-LIKE RECEPTOR 4

Y. Liu1, W. Jiang2. 1Gastroenterology, 2Zhongshan Hospital Fudan

University, Shanghai, China

E-mail: liuyun2008294@163.com

Background and Aims: Here we evaluated the therapeutic effects

of 3, 3’-diindolylmethane (DIM), which is extracted from cruciferous

vegetables, on the progression of ConA-induced liver fibrosis, and

further investigated the possible immunoregulatory mechanism.

Methods: Mice were administrated with ConA together with

or without DIM for 8 weeks. The subtypes of hepatic and

splenic CD4+T cells were tested by flow cytometry and

immunohistochemistry. The detection of cytokines were measured

by ELISA. The protein expressions of CYP1A1, CYP1B1 and TLR4

were tested by western blot, and AhR antagonist CH223191 or anti-

TLR4 neutralizing antibody was used in vitro to testify the possible

immunoregulatory signaling of DIM.

Results: DIM significantly alleviated ConA-induced hepatic injury,

inflammation and fibrosis. DIM also inhibited the proliferation

of HSCs and pro-fibrotic cytokines secretion. The treatment with

DIM significantly led to the induction of Tregs, while reciprocally

reducing Th17 populations. And DIM also modulated associated

S266 Journal of Hepatology 2014 vol. 60 | S215–S359