Supplementary Materials
Patients’ information.
The inclusion criteria for participants are: (i) gastric adenocarcinoma with a pathological diagnosis; (ii)
informed consent or waiver of consent provided by the patient; and (iii) follow-up information
available. We excluded GC patients with (i) failure to provide informed consent; (ii)
non-adenocarcinoma or multiple cancers; (iii) no tissue sample obtained; (iv) loss of contact after
surgery; or (v) stage IV GC without palliative surgery. Among the 311 GC patients, 237 cases are with
R0 resection, 63cases with R1 resection and 11 cases with R2 resection. Patients were obtained their
surgical operation during 1997 to 2001. All GC patients were Asian (Chinese). 153 of 311 patients had
post-surgery adjuvant chemotherapy. The combination chemotherapy regimens included folinic acid,
5-fluorouracil and oxaliplatin (FOLFOX6; 73 Cases); epirubicin, oxaliplatin and Xeloda (EOX; 12
cases); 5-fluorouracil, epidoxorubicin and mitomycin C (FEM; 9 cases); etoposide, leucovorin and
5-fluorouracil (ELF; 38 cases); mitomycin C and 5-fluorouracil (MF; 4 cases); and others (oral S-1/x,
docetaxel-based and other protocols; 17 cases). All patients were followed up until January 2012. The
TNM stage data for the participants were obtained from the clinical and pathological diagnoses and
determined according to the NCCN guidelines for GC (Version 2, 2011).
Table S1. LZTFL1 expression was associated with GC cell differentiation
Cell lines name
MKN28
MKN45
SGC7901
MC803
Normal immortal gastric
epithelium
Middle differentiation
Low differentiation
Low differentiation
Low differentiation
Low differentiation
Low differentiation
KatoIII
NCI-N87
HGC27
Signet-ring cell
Liver metatsasis
Undifferentiation
GES 1
BGC823
AGS
LZTFL1 expression
level
Differentiation/Origination
High
High
Moderate
Low
Low
Low
Low
Scarce
Scarce
Scarce
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Table S2. siRNA sequences used in this work
Name
Sequence
Sense 5’UUCUCCGAACGUGUCACGUTT3’
Si-control
Antisense 5’ACGUGACACGUUCGGAGAATT3’
Sense 5’GCAGAGAUUACAUCUUCAATT3’
Si-LZTFL1-homo-496
Antisense 5’UUGAAGAUCUAAUCUCUGCTT3’
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Table S3. Primers used in this work and their sequences
Primers names
Tm(oC)
Sequences
AGATTCCTGCTTCCAAGAC
55.4
CTGCTGTTCCACCTTCAT
55
GTCTCTCTCACCACCTCCACAG
63.8
CTCGGACACTTCCACTCTCTTT
60.07
TATCAACCAGAGGGAGTG
61.92
GCGAGGAGAGCAGGATTTCT
59.85
AGAGACAGTGGATGATGCCTTT
58.2
ATCGTCATCAAAATGGGAGTCT
56.3
TGTACCGCTATGGTTACACTCG
60.1
GGCAGGGACAGTTGCTTCT
59.7
TTTCTGGTTCTGTGTCCTCTG
58
TGTCAGCCTTTGTCCTGTAGC
60
CTGGCTGGTGGAAATGATGT
57.6
AAGGACGGCTTCTTCTCTTATG
58.2
CAGGAAGAACCCTTGAACTTGT
58.2
GGCACTTGGTGGGATTACATT
58
TCCGAAGCCAAATGACAAA
53.2
CTCTCTCTGTGGGTGTGTTGT
61.9
GAGCAAGATTCAGACCCTCAAG
60.1
CCATCCTCCAGACCGAGAAG
61.9
GAAGGTGAAGGTCGGAGTCA
60
TTCACACCCATGACGAACAT
60
LZTFL1
E-Cadherin
Vimentin
MMP2
MMP9
Snail
ZO-1
ZEB-1
Slug
Twist
GAPDH
3
Figure S1. Alteration o LZTFL1 in SGC7901 or BGC823 cells does not change the cellular
morphology. (A) SGC7901 cells were transfected without, with pcDNA, and with pcDNA-LZTFL1.
(B) BGC823 cells were transfected without, with control and LZTFL1 siRNA.
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Figure S2. LZTFL1 inhibits cellular phenotypes associated with EMT. (A) Migration assays of
SGC7901 and BGC823 cells transfected with/without indicated plasmid or siRNA were measured by
Transwell assay. Representative images for SGC7901 and BGC823 cells were shown. Cells migrated
to the membrane facing the lower-chamber were stained with DAPI. (B) The invasion of cells
described in (A) across matrigel-coated transcwell plates was assayed. Migrated cells were stained
with 0.5% crystal violet.
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