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Page 35 of 36 J Cancer Metastasis Treat 2019;5:31 I http://dx.doi.org/10.20517/2394-4722.2019.21

DEGs identified by this methodology, NRN1 was selected for further analyzes. This study aimed to
evaluate, compare and correlate NRN1 mRNA and methylation levels of GC and adjacent non tumor
samples from patients with primary gastric adenocarcinoma. The mRNA level was assessed by quantitative
reverse transcription PCR. DNA methylation analysis was assessed by Ion Torrent PGM sequencer. A total
of 12 CpG sites were evaluated in NRN1 promoter region. GC samples showed a significant increased
DNA methylation at five specific CpG sites of NRN1 promotor region compared with adjacent non tumor
samples (P < 0.05), besides no difference in NRN1 mRNA levels between these groups. At this time, no
significant correlation between mRNA and methylation levels, as well as, significant clinicopathological
associations were observed. Although different mechanisms may be involved in gene regulation, NRN1
methylation seems to play an important role in gastric carcinogenesis. This study is still in progress and
new samples and other gene regions will be incorporated in our analyzes.

Biography
Fernanda Wisnieski has completed her PhD at Federal University of São Paulo (UNIFESP), São Paulo,
Brazil. Currently she is a postdoctoral researcher at Genetics Division of UNIFESP. She has experience in
the area of Human and Medical Genetics, working mainly on genetics and epigenetics aspects of gastric
cancer clinical samples and cell lines models. Since the end of her doctorate degree she has published 15
papers in the area.

46. A semi-comprehensive analysis of gene amplification in breast cancers using multiplex

ligation-dependent probe amplification and fluorescence in situ hybridization

Akishi Ooi

Kanazawa University, Kanazawa 920-1192, Japan.

Gene amplification is a common event in breast cancer. The aim of the present study was to determine the
amplification status of 22 genes that are reportedly frequently amplified in breast cancers. A total of 320
formalin-fixed and paraffin-embedded invasive ductal cancer tissues were screened by multiple ligation-
dependent probe amplification, and 885 genes with “gain” or “amplified” status were further examined for
the respective gene amplification by fluorescence in situ hybridization (FISH). In the FISH analysis, 116
of 320 tumors (36%) displayed gene amplification of at least 1 of the 22 genes. The frequencies of gene
amplification were as follows: ESR1 (5 cases, 1.5%), EGFR (one case, 0.3%), ZNF703 (25 cases, 7.8%), FGFR1
(24 cases, 7.5%), ADAM9 (17 cases, 5.3%), IKBKB (10 cases, 3.1%), PRDM1 (5 cases, 1.5%), MTDH (15 cases,
4.7% ), MYC (29 cases, 9.0%), CCND1 (39 cases, 12%), C11ORF30 (19 cases, 5.9%), CDH1 (1 case, 0.3%),
CPD (9 cases, 2.8%), MED1 (25 cases, 7.8% ), ERBB2 (35 cases, 11%), CDC6 (14 cases, 4.3%), TOP2A (12
cases, 3.8%), MAPT (1 case, 0.3%), PPM1D (10 cases, 3%), BIRC5 (6 cases, 1.9%), CCNE1 (3 cases, 0.9%),
and AURKA (1 case, 0.3%). In addition to the frequent co-amplifications of genes closely located on the
same chromosome region, such as 8p11, 8q24, 11q13, or 17q12-21, co-amplification of genes located
in different chromosomes was also found. The co-amplification of ZNF703/FGFR1 (8p11) and CCND1
(11q13) in single cells was found in 11 tumors, occurring in the same amplicons in 5. The co-amplification
of ZNF703/FGFR1 and ERBB2 (17q12-21) in single tumor nuclei but different amplicons was observed in
11 cases, and that of CCND1 and ERBB2 was noted in 5 cases. The co-amplification of MYC (8q24) with
CCND1 or ERBB2 in single nuclei was found in three and two tumors, respectively. The amplifications of
ESR1 and ERBB2 as well as EGFR and ERBB2 were found in one tumor each, although in different cancer
cells. Amplified genes are attractive potential targets for therapy, so a precise and feasible analysis of the
gene amplification status is clinically valuable.