Page 130 - ebook HCC

EASL
PROGRAMME
GENEVA, SWITZERLANDA, SWITZERLAND
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128 PROGRAMME AND ABSTRACTSAND ABSTRACTS GENEV EASL HCC SUMMITHCC SUMMIT 129
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FEBRUAR
FEBRUARY 13 - 16, 2014Y 13 - 16, 2014
Poster Board Number B24 Poster Board Number B25
MICRORNA-21 STIMULATES CANCER GROWTH
HUMAN BILE CONTAIN MICRORNA-LADEN AND INVASION BY INHIBITING THE TUMOR
EXOSOMES THAT CAN BE USED FOR CANCER SUPPRESSOR EFFECTS OF SERPINI-1, PDCD4
DIAGNOSIS AND PTEN

Ciprian Tomuleasa , Florin Selaru 2 Tanase Timis , Ciprian Tomuleasa and HCC Working group
1
1
1
2
1 Chemotherapy, Ion Chiricuta Cancer Center, Cluj Napoca, Romania, Medicine, 3The 1 Chemotherapy, Ion Chiricuta Cancer Center, Cluj Napoca, Romania
Johns Hopkins University School of Medicine, Baltimore, United States
Corresponding author’s e-mail: timis.tanase@gmail.com
Corresponding author’s e-mail: ciprian.tomuleasa@umfcluj.ro
Introduction: Studies revealed that aberrant microRNAs play vital roles in oncogenesis
Introduction: Hepatocellular carcinoma (HCC) presents significant diagnostic challenges, via regulation of various gene expression or protein translation. MicroRNA-21 (miR-21) is
resulting in late patient diagnosis and poor survival rates. Consequently, significant efforts abnormally expressed in many solid cancers such as liver adenocarcinoma and regulates
are needed to develop improved diagnostics. MicroRNAs (miRs) have recently emerged targets involved in cancer initiation and progression.
as a valuable class of diagnostic markers; however, thus far, neither exosomes nor miRs
within exosomes have been investigated in human bile. Aims: We investigated the function of miR-21 in liver cancer, and it’s potential targeting
of the tumor suppressor genes Serpini-1, phosphatase and tensin homolog (PTEN) and
Aims: We aimed to comprehensively characterize human biliary exosomes, including
programmed cell death protein 4 (PDCD4).
BASIC POSTER ABSTRACTS Methodology: Human bile specimens were obtained from patients under a Hopkins Methodology: After using quantitative RT-PCR in order to verify the overexpression of BASIC POSTER ABSTRACTS
their miR content.
miR-21 in two hepatocellular carcinoma cell lines. Western blotting confirmed the RT-
Institutional Review Board (IRB) approval. Specimens were processed for exosome
PCR data in a set of rescue experiments that microRNA-21 mimic, inhibitor, negative and
isolation. The exosomes were analyzed with western blotting, transmission electron
positive control. The protein levels of miR-21 targets Serpini-1, PTEN and PDCD4 was
microscopy (TEM) and nanoparticle tracking analysis (NTA). Multivariate Organization of
Combinatorial Alterations (MOCA) was utilized to analyze the data and develop a miR-
two different cell lines.
based HCC diagnostic panel. measured. Afterwards, we evaluated it’s effect on tumor growth and invasion potential on
Results: RT-PCR results proved that miR-21 is overexpressed in hepatocellular carcinoma
Results: We have established the presence of exosomes in human bile. In addition, we cells when compared with hepatic cells. Western blot results further suggest that PTEN
have demonstrated that human biliary exosomes contain abundant miR species, which are and PDCD4 are regulated by miR-21, as miR-21 inhibitor increases the expression of
stable and therefore amenable to the development of disease marker panels. Furthermore, PTEN and PDCD4 proteins and significantly reduces cell proliferation, migration and
we have characterized the protein content, size, numbers and size distribution of human invasion. The control experiment proved that the miR-21 mimic significantly inhibits the
biliary exosomes. Finally, utilizing MOCA, we defined a novel biliary exosomal miR-based expression of PTEN and PDCD4 proteins, leading to an increase in cell invasion and
panel for HCC diagnosis which demonstrated a sensitivity of 60% and specificity of 93%. migration. Furthermore, and miR-21 inhibitor inhibits the apoptosis of cancer cell lines.

Conclusions: This study reports the results of a comprehensive characterization of Conclusions: MicroRNA-21 is overexpressed in hepatocellular carcinoma and it’s
human biliary exosomes, including their miR content. In addition, these findings establish aberrant expression may have important roles in cancer growth and dissemination by
the importance of using exosomes, rather than whole bile, for developing miR-based modulating the expression of the tumor suppression genes PTEN and PDCD4, as well
disease markers in bile. The novel HCC panel developed herein represents a valuable as the pathways involved in mediating cell growth, migration, invasion and apoptosis.
addition to current diagnostic methods. Targeting of miR-21 may help us to develop novel therapeutics for liver cancer.