Radiol Oncol 2024; 58(2): 234-242. doi: 10.2478/raon-2024-0020
234
research article
Correlation of laminin subunit alpha 3 
expression in pancreatic ductal 
adenocarcinoma with tumor liver metastasis 
and survival
Yueyi Xing1, Xue Jing2, Gong Qing1, Yueping Jiang2
1 Qingdao University, Qingdao, Shandong Province, China
2 Gastroenterology Department, the Affiliated Hospital of Qingdao University, Qingdao, Shandong Province, China
Radiol Oncol 2024; 58(2): 234-242.
Received 30 October 2023 
Accepted 14 January 2024
Correspondence to: Dr. Yueping Jiang, Gastroenterology Department, the Affiliated Hospital of Qingdao University, Qingdao, Shandong 
Province, China. E-mail: yuepingmd@hotmail.com
Disclosure: No potential conflict of interest were disclosed.
This is an open access article distributed under the terms of the CC-BY license (https://creativecommons.org/licenses/by/4.0/).
Background. The high mortality rate of pancreatic ductal adenocarcinoma (PDAC) is primarily attributed to me-
tastasis. Laminin subunit alpha 3 (LAMA3) is known to modulate tumor progression. However, the influence of LAMA3 
on liver metastasis in PDAC remains unclear. This study aimed to elucidate whether LAMA3 expression is increased in 
PDAC with liver metastasis.
Patients and methods. We extracted information related to LAMA3 expression levels and associated clinicopatho-
logical parameters from The Cancer Genome Atlas (TCGA) and four Gene Expression Omnibus (GEO) datasets. 
Clinicopathological analysis was performed; the Kaplan-Meier Plotter was used to evaluate LAMA3’s prognostic ef-
fect in PDAC. We retrospectively collected clinicopathological data and tissue specimens from 117 surgically treated 
patients with PDAC at the Affiliated Hospital of Qingdao University. We assessed LAMA3 expression and investigated 
its correlation with the clinicopathological traits, clinical outcomes, and hepatic metastasis.
Results. Amplified expression of LAMA3 was observed in PDAC tissue compared with normal tissue in the TCGA and 
GEO databases. High LAMA3 expression was associated with poor overall survival (OS) and relapse-free survival (RFS) 
in patients with PDAC. LAMA3 expression was significantly enhanced in PDAC tissues than in adjacent tissues. Tumor 
tissues from patients with PDAC exhibiting liver metastasis showed higher LAMA3 expression than those without liver 
metastasis. High LAMA3 expression correlated with large tumor size and TNM stage. LAMA3 expression and liver me-
tastasis were independent predictive factors for OS; the former was independently associated with liver metastasis.
Conclusions. LAMA3 expression is elevated in patients with PDAC with liver metastasis and is a predictor of prognosis.
Key words: pancreatic ductal adenocarcinoma; laminin subunit alpha 3; liver metastasis; prognosis
Introduction
Pancreatic ductal adenocarcinoma (PDAC), a dis-
ease that is prevalently observed within the diges-
tive system, is distinguished by its severe malig-
nancy and exhibits a disconcerting confluence of 
incidence and mortality.1 The 5-year survival rate 
of patients with PDAC is < 10%, with an extremely 
poor prognosis. If this trend is sustained, the im-
pending decade may witness pancreatic cancer 
ascending to the rank as the second most lethal 
cancer.2 Most patients with PDAC remain asymp-
Radiol Oncol 2024; 58(2): 234-242.
Xing Y et al./ Laminin subunit alpha 3 in pancreatic ductal adenocarcinoma 235
tomatic until the disease reaches advanced stages. 
Ninety percent of patients with PDAC diagnosed 
only after metastasis have a poor prognosis, with 
50% developing systemic metastasis.3,4 The poten-
tial for enduring survival among patients with 
PDAC considerably depends on tumor size and 
disease stage. Therefore, early detection of po-
tentially curable cancers is crucial for reducing 
mortality rates among patients with PDAC. The 
elucidation of key molecular mechanisms and pro-
spective intervention targets associated with pan-
creatic cancer metastasis will aid in deciphering 
the genetic and molecular underpinnings of this 
disease, provide biomarkers for preliminary warn-
ing and metastasis surveillance, and pave the way 
for enhancing the survival prospects of patients 
with pancreatic cancer.
Laminin, a heterotrimeric molecule consisting 
of α, β, and γ subunits, is the primary constituent 
of the extracellular matrix while collagen and fi-
bronectin form the basement membrane. Among 
the three subunits of laminin, the α subunit is in-
volved in tissue-specific distribution and biologi-
cal activity.5 Laminin subunit alpha 3 (LAMA3), 
which encodes for the laminin α subunit, enables 
its globular carboxyl-terminal domain to engage 
with integrins at the plasma membrane, thereby 
participating in intracellular signal transduction.6 
Currently, LAMA3 contributes to cell prolifera-
tion and apoptosis in diverse malignant tumors 
and modulates tumor progression through signal 
transduction pathways, such as focal adhesion 
plaques.7-9 The aberrant expression of LAMA3 in 
various tumors is inextricably associated with the 
clinical stage, tumor size, and pathological mani-
festations of patients.10 However, the influence of 
LAMA3 on liver metastasis in PDAC remains un-
clear. This study aimed to clarify LAMA3 expres-
sion in PDAC and investigate the relationship be-
tween LAMA3 expression and liver metastasis in 
patients with unresectable PDAC.
Patients and methods
Procurement of bioinformatics analysis 
data
RNA sequencing expression traits, along with their 
associated clinical data pertaining to LAMA3, 
were procured from the The Cancer Genome Atlas 
(TCGA) dataset (https://portal.gdc.com). The cur-
rent-release GTEx datasets were accessed from the 
GTEx data portal website (https://www.gtexportal.
org/home/datasets). The data comprised 179 tumor 
samples and 4 normal samples sourced from the 
TCGA, in addition to 328 normal mRNA expres-
sion data points from GTEx. Corresponding plat-
form annotation files were obtained from the Gene 
Expression Omnibus (GEO) database (http://www.
ncbi.nlm.nih.gov/geo/) to validate the expression 
levels of LAMA3 in PDAC. Finally, we identified 
four datasets: GSE28735 (n = 90), GSE62452 (n = 
130), GSE101448 (n = 43), and GSE62165 (n = 131). To 
perform a clinicopathological analysis of LAMA3, 
we used University of ALabama at Birmingham 
CANcer (UALCAN) (http://ualcan.path.uab.edu). 
Survival curves were generated using the Kaplan-
Meier Plotter database (http://kmplot.com/analy-
sis/).11
Acquisition of human pancreatic ductal 
adenocarcinoma (PDAC) specimens and 
clinicopathological data
Our study included 117 patients with PDAC who 
underwent pancreatic surgery at the Affiliated 
Hospital of Qingdao University. These patients 
had not received any anticancer treatment before 
surgery, and the diagnosis of pancreatic carci-
noma was confirmed by postoperative pathol-
ogy. Paraffin-embedded tumor tissues were ob-
tained from each patient, and the corresponding 
para-carcinomatous tissues were obtained from 
60 patients. All patients provided informed con-
sent, and the investigation was conducted in ac-
cordance with the Declaration of Helsinki with 
the endorsement of the Medical Ethics Committee 
of the Affiliated Hospital of Qingdao University 
(QYFYWZLL27485 and QYFYWZLL27608).
Clinicopathological data were obtained from 
retrospective medical records, which consisted 
of age, sex, tumor size, tumor location, histo-
logical grade, perineural invasion, lymph node 
metastasis, vascular invasion, liver metastasis, 
tumor-node-metastasis (TNM) stage, preopera-
tive serum carcinoembryonic antigen (CEA), and 
carbohydrate antigen 19-9 (CA19-9) concentra-
tions. Overall survival (OS) was calculated as the 
interval between surgery and either death or last 
follow-up appointment. The dates of death were 
ascertained from hospital records or follow-up tel-
ephone interviews.
Immunohistochemistry
Paraffin-embedded PDAC and para-cancerous tis-
sues underwent sequential sectioning at a thick-
ness of 4 μm. After baking, deparaffinizing, and 
Radiol Oncol 2024; 58(2): 234-242.
Xing Y et al./ Laminin subunit alpha 3 in pancreatic ductal adenocarcinoma236
hydrating, the paraffin sections were ensconced 
in a pressure cooker for 10 min for antigen repair. 
Subsequently, the antigen repair box was relocated 
to an ice box for a 25-min interval, permitting cool-
ing to room temperature. To curb endogenous per-
oxidase activity, the tissue sections were immersed 
in a concoction of 3% hydrogen peroxide and 
methanol for 15 min. Each section received a block-
ade of 10% sheep serum and incubated at 37°C for 
half an hour. This was followed by an overnight 
incubation at 4°C with primary antibodies (1:100 L, 
no. ab242197; Abcam Inc.), followed by incubation 
with secondary antibodies (no. ab242197; Abcam 
Inc.) at 37°C for 30 min. The tissue sections were 
then stained with 3, 3-diaminobenzidine (Roche) 
for 5–10 min at room temperature. Hematoxylin 
(Roche) was used for counterstaining for 25 s be-
fore proceeding with dehydration, clarification, 
and sealing. Microscopic visualization was per-
formed to record the images. An independent duo 
of pathologists evaluated all samples.
The cytoplasmic staining score (CF) was de-
fined as follows: 0 (0–20%), 1 (21–50%), 2 (51–75%), 
and 3 (>75%). Moreover, the cytoplasmic staining 
intensity (CI) was categorized as 0 (negative), 1–2 
(weak), and 3 (strong). The cytoplasmic composite 
score was calculated as CF×CI.
Statistical analyses
For all the TCGA and GEO databases, we used 
the Wilcox test to perform differential expres-
sion analysis between tumor and normal tissues. 
Categorical variables are expressed as frequencies 
and percentages, and significance was determined 
using the χ² or Fisher’s exact test. Quantitative 
variables are expressed as means±standard de-
viations, and significance was determined using 
Student’s t-test. Non-normally distributed vari-
ables are expressed as medians and interquartile 
ranges, and significance was determined using 
the Mann–Whitney U test. Multivariate logistic 
regression analyses were performed to identify 
the independent risk factors for PDAC. We used 
the cutoff points of the test variables produced on 
receiver operating characteristic curves. Survival 
analysis was performed using Kaplan–Meier 
analysis and assessed using the log-rank test. Cox 
regression analysis was performed to analyze the 
effect of OS on the survival of patients with PDAC. 
All analyses were performed using SPSS version 
24.0, GraphPad Prism version 8.0.1, and R software 
version 4.0.3. P < 0.05 was considered statistically 
significant.
Results
Elevation of laminin subunit alpha 3 in 
PDAC and its correlation with prognosis 
using bioinformatics analysis
Using the TCGA database, we identified a promi-
nent divergence in LAMA3 expression between 
PDAC tissues (n = 179) and normal tissues (n = 332) 
FIGURE 1. Expression of laminin subunit alpha 3 (LAMA3) in pancreatic ductal 
adenocarcinoma (PDAC) and normal tissues from the Cancer Genome Atlas 
(TCGA) database (A) and the Gene Expression Omnibus (GEO) database (B-E). 
Expression of LAMA3 in 179 PDAC and 332 normal tissues from TCGA (A); Expression 
of LAMA3 in 45 PDAC and 45 normal tissues from GSE28735 cohort (B); Expression 
of LAMA3 in 69 PDAC and 61 normal tissues from GSE62452 cohort (C); Expression 
of LAMA3 in 24 PDAC and 19 normal tissues from GSE101448 cohort (D); Expression 
of LAMA3 in 118 PDAC and 13 normal tissues from GSE62165 cohort (E). 
****P < 0.001.
A
B C
D E
Radiol Oncol 2024; 58(2): 234-242.
Xing Y et al./ Laminin subunit alpha 3 in pancreatic ductal adenocarcinoma 237
(P < 0.001) (Figure 1A). Four datasets (GSE28735, 
GSE62452, GSE101448, and GSE62165) were ob-
tained from the GEO database and used as vali-
dation sets. The results showed that LAMA3 was 
significantly upregulated in PDAC tissues (all P < 
0.001) (Figure 1B-E).
To further elucidate the role of LAMA3 in PDAC, 
we investigated its expression using various clin-
icopathological parameters. LAMA3 expression 
displayed no remarkable correlation with age 
(Supplementary Figure 1A), sex (Supplementary 
Figure 1B), and drinking habits (Supplementary 
Figure 1C) in patients with PDAC. Grade 1 indicat-
ed a well-differentiated (low-grade) tumor, grade 
2 denoted a moderately differentiated (intermedi-
ate-grade) tumor, grade 3 indicated a poorly differ-
entiated (high-grade) tumor, and grade 4 indicated 
an undifferentiated (high-grade) tumor. The grade 
of patients with PDAC influenced LAMA3 expres-
sion, and heightened expression was observed 
in grades 2 and 3 (Supplementary Figure 1D). 
However, there was no significant difference in 
LAMA3 expression with respect to nodal me-
tastasis (Supplementary Figure 1E) or diabetes 
(Supplementary Figure 1F).
Survival curves were generated using the 
Kaplan–Meier Plotter database. Elevated LAMA3 
expression was positively associated with poorer 
OS (Figure 2A, hazard ratio [HR] = 3.86, P < 0.001) 
and relapse-free survival (RFS) (Figure 2B, HR = 
406726946.65, P < 0.001). These results demonstrate 
that high LAMA3 expression is associated with an 
unfavorable prognosis in patients with PDAC.
Clinical characteristics of patients with 
PDAC
After meticulous filtering according to the inclu-
sion and exclusion criteria, 117 patients with PDAC 
were included in this study. The baseline charac-
teristics of the patients are summarized in Table 1. 
The mean age of all patients was 62.43 ± 9.33 years, 
with males accounting for 73 (62.3%) of the total 
population. Pancreaticoduodenectomy and dis-
tal pancreatectomy were performed in 69 and 48 
patients, respectively. In our cohort, 61 postsur-
gical patients received chemotherapy, 3 patients 
received radiotherapy, 6 patients received immu-
notherapy, and 4 patients received interventional 
therapy. Based on the immunohistochemical re-
sults of LAMA3 expression levels, the patients 
were categorized into groups with high or low 
expression. Notably, 62 patients showed elevated 
LAMA3 expression. The univariate analyses of the 
two cohorts are presented in Table 1.
Increased LAMA3 expression correlated with 
large tumor size (P = 0.007), and the degree of 
LAMA3 expression was associated with different 
TNM stages (P = 0.002). In addition, LAMA3 ex-
pression was higher in tumor tissue from patients 
with PDAC and liver metastases than those with-
out liver metastases (P = 0.005). In the two groups, 
the surgical modalities used were significantly 
different (P = 0.036), but there were no significant 
differences in age, gender, tumor location, histo-
logical grade, perineural invasion, vascular inva-
A B
C D
FIGURE 2. The expression of laminin subunit alpha 3 (LAMA3) for prediction of 
overall survival (OS) and relapse free survival (RFS) in patients with pancreatic 
ductal adenocarcinoma (PDAC). OS (A), RFS (B).
A B
FIGURE 3. Representative immunohistochemical staining of laminin subunit alpha 
3 (LAMA3) in pancreatic ductal adenocarcinoma (PDAC) and adjacent normal 
tissue. High expression of LAMA3 in PDAC tissue (A) compared with adjacent tissue 
(B). Low expression of LAMA3 in PDAC tissue (C) compared with adjacent tissue 
(D). Magnification, x400
Radiol Oncol 2024; 58(2): 234-242.
Xing Y et al./ Laminin subunit alpha 3 in pancreatic ductal adenocarcinoma238
sion, lymph node metastasis, CEA and CA19-9 lev-
els, and adjuvant systemic therapy (P > 0.05).
Heightened expression of LAMA3 in 
PDAC tissues relative to adjacent tissues
Immunohistochemistry was performed to meas-
ure LAMA3 expression in PDAC and adjacent 
normal tissues. LAMA3 staining was almost un-
detectable in normal tissues, and protein intensity 
was negative (Supplementary Table 1, Figure 3). 
Conversely, moderate staining and a robust inten-
sity of LAMA3 protein expression were observed 
in PDAC tissues. The results demonstrated that 
LAMA3 expression was significantly higher in 
carcinoma specimens than in the adjacent tissues 
(P < 0.001).
Superior expression of LAMA3 in PDAC 
tumor tissues exhibiting liver metastasis
All patients with PDAC were categorized into two 
groups based on the emergence or absence of liver 
metastasis postoperatively (Table 2). Univariate 
analysis showed that histological grade (P = 0.001), 
TNM stage (P = 0.013), and vascular invasion (P 
= 0.018) were significantly associated with liver 
metastasis. Immunohistochemistry was used to 
assess LAMA3 expression in patients with PDAC 
with or without liver metastasis. LAMA3 expres-
sion in tumor tissues from patients with PDAC 
and liver metastasis was significantly higher 
than in those without liver metastasis (P = 0.005). 
Representative immunohistochemical images are 
shown in Figure 4. Age, sex, tumor location, tu-
mor size, lymph node metastasis, perineural in-
vasion, and serum CEA and CA19-9 levels were 
not associated with the development of liver me-
tastasis. Significant factors from the univariate 
analysis, as shown in Table 2, were incorporated 
into the multivariate logistic regression analysis 
(Supplementary Table 2). The results showed that 
histological grade and LAMA3 expression were 
independently associated with liver metastasis.
High LAMA3 expression correlates with 
poor PDAC prognosis
The median survival times of patients with low 
and high LAMA3 expressions were 29 and 14 
months, respectively. The 1-, 2-, and 3-year surviv-
al rates of the high-expression group (n = 62) were 
58.1%, 14.5%, and 4.8%, respectively. Conversely, 
those in the low-expression group (n = 55) were 
90.9%, 47.2%, and 16.4%, respectively. Using 
TABLE 1. Characteristics of all patients
Characteristics All
LAMA3 expression
High  
(n = 62)
Low  
(n = 55) P-value
Age(year), mean±SD 62.43 ± 9.33 62.19 ± 9.38 62.69 ± 9.36 0.775
Sex, n (%) 0.615
   Male 73 (62.4) 40 33
   Female 64 (37.6) 22 22
Tumor location, n (%) 0.054
   Head 70 (59.8) 32 38
   Body and tail 47 (40.2) 30 17
Tumor size, n (%) 0.007
   ≤ 2 cm 5 (4.3) 2 3
   > 2 cm and ≤ 4 cm 81 (69.2) 37 44
   > 4 cm 31 (26.5) 23 8
Histological grade, n (%) 0.810
   G1 31 (26.5) 17 14
   G2–3 86 (73.5) 45 41
TNM stage, n (%) 0.002
   I–II 92 (78.6) 42 50
   III–IV 25 (21.4) 20 5
Perineural invasion, n (%) 0.921
   Yes 91 (77.8) 48 43
   No 26 (22.2) 14 12
Vascular invasion, n (%) 0.340
   Yes 37 (31.6) 22 15
   No 80 (68.4) 40 40
Lymph node metastasis, n (%) 0.255
   Yes 49 (41.9) 29 20
   No 68 (58.2) 33 35
Liver metastasis, n (%) 0.005
   Yes 45 30 15
   No 72 29 43
CEA (ng/ml) 0.392
   ≤ 12 108 (92.3) 56 52
   > 12 9 (7.7) 6 3
CA19-9 (U/ml) 0.395
   ≤ 282 74 (63.2) 37 37
   > 282 43 (36.8) 25 18
Surgical modalities, n (%) 0.036
   Pancreaticoduodenectomy 69 (59) 31 38
   Distal pancreatectomy 48 (41) 31 17
Postoperative 
chemotherapy, n (%) 0.77
   Yes 60 40 20
   No 57 41 16
Postoperative radiotherapy, 
n (%) 0.063
   Yes 3 2 1
   No 114 79 35
Immunotherapy, n (%) 0.068
   Yes 6 4 2
   No 111 77 34
Interventional therapy, n (%) 0.374
   Yes 4 4
   No 114 77 36
CEA = carcinoembryonic antigen; CA19-9 = carbohydrate antigen 19-9; LAMA3 = laminin 
subunit alpha 3
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Xing Y et al./ Laminin subunit alpha 3 in pancreatic ductal adenocarcinoma 239
Kaplan–Meier curves, high LAMA3 expression in 
PDAC was associated with poor OS, suggesting an 
unfavorable prognosis (P < 0.001) (Figure 5).
Univariate analysis suggested that tumor size, 
TNM stage, liver metastasis, and LAMA3 expres-
sion had a significant prognostic influence on OS 
(Table 3). Multivariate survival analysis revealed 
that LAMA3 expression (HR, 2.016; 95% confi-
dence interval [CI], 1.257–3.234; P = 0.004) and liv-
er metastasis (HR, 2.284; 95% CI, 1.426–3.657; P = 
0.001) were independent predictive factors of OS.
Discussion
Pancreatic cancer is a highly aggressive neoplasm 
of the digestive system and is characterized by a 
mortality rate equal to its incidence rate. Its strong 
invasiveness and early metastasis render approxi-
mately 80% of patient ineligible for surgical inter-
vention at the time of diagnosis. This results in 
a 5-year survival rate < 10%. Current treatment 
approaches for PDAC include surgical resection 
combined with chemotherapy, radiation therapy, 
interventional therapy, and immunotherapy. Even 
in patients with resectable localized tumors, the 
postoperative 5-year survival rate remains ap-
proximately 20%.12 The major contributor to the 
high mortality rate of PDAC is its propensity for 
early metastasis, which poses a significant chal-
lenge in clinical management. Consequently, there 
is an urgent need to identify additional predictive 
biomarkers to enhance the risk stratification of pa-
tients with PDAC.
LAMA3, a member of the laminin family, plays 
a pivotal role in cellular processes by interacting 
with integrins on the cell membrane and par-
ticipating in the intracellular signal transduction 
pathways. Recent studies have implicated elevated 
LAMA3 expression in various types of tumors, 
where it appears to promote cell proliferation, 
apoptosis, and tumor progression by modulating 
signal transduction pathways.7-9 Zboralski et al. 
demonstrated the simultaneous functional inac-
tivation of the tumor suppressor mothers against 
decapentaplegic homolog 4 (SMAD4) and invasive 
growth of tumors in rectal and pancreatic cancers. 
Laminin 332 (LM-332) is the target of SMAD4, a 
positive transcriptional regulator that promotes 
the transcription of LAMA3, LAMB3, and LAMC2 
genes encoding LM-332. SMAD4 mediates tran-
scriptional activity through distinct molecular 
mechanisms associated with the LAMA3, LAMB3, 
and LAMC2 promoters.13 Additionally, Huang et 
al. highlighted the increased expression of LAMA3 
protein in PDAC tumor cells relative to that in nor-
mal pancreatic cells. They further showed that 
high LAMA3 expression promoted the prolifera-
tion, migration, and invasion of PDAC tumor cells.5 
However, the effect of LAMA3 on liver metastasis 
in PDAC has not been fully elucidated. This study 
aimed to clarify the expression profile of LAMA3 
in PDAC and investigate its potential association 
with liver metastasis in patients diagnosed with 
unresectable PDAC.
In this study, we integrated data from the TCGA 
database with four independent datasets from the 
GEO database as validation cohorts and found that 
LAMA3 expression was upregulated in PDAC, 
validating previous study findings.14 Furthermore, 
through the analysis of clinical samples, we ob-
A B
FIGURE 4. Representative immunohistochemical staining of laminin subunit alpha 
3 (LAMA3) in pancreatic ductal adenocarcinoma (PDAC) with and without liver 
metastasis. Low expression of LAMA3 in PDAC tissues (A) without liver metastatic. 
High expression of LAMA3 in PDAC tissues (B) with liver metastasis. Magnification, 
x400.
FIGURE 5. The expression of laminin subunit alpha 3 (LAMA3) for prediction of 
overall survival (OS) in patients with Pancreatic ductal adenocarcinoma (PDAC). 
Survival analysis was carried out with Kaplan-Meier and checked by log-rank test.
Radiol Oncol 2024; 58(2): 234-242.
Xing Y et al./ Laminin subunit alpha 3 in pancreatic ductal adenocarcinoma240
served that LAMA3 was overexpressed in pan-
creatic carcinoma tissues compared with adjacent 
noncancerous tissues. When we analyzed clinico-
pathological data from the UALCAN database, we 
TABLE 2. Univariate analysis of clinicopathological characteristics in patients with 
pancreatic ductal adenocarcinoma with and without liver metastasis
Characteristics Liver metastasis (n = 45)
Without liver 
metastasis (n = 72) P-value
Age (year), M (IQR) 63 (58–69) 63 (56–68) 0.814
Sex, n 0.717
   Male 29 44
   Female 16 28
Tumor location, n 0.456
   Head 25 45
   Body and tail 20 27
Tumor size, n 0.240
   ≤ 2 cm 2 3
   > 2 cm and ≤4 cm 28 53
   > 4 cm 15 16
Histological grade, n 0.001
   G1 20 11
   G2-3 25 61
TNM stage, n 0.013
   I–II 30 62
   III–IV 15 10
Perineural invasion, n 0.170
   Yes 38 53
   No 7 19
Vascular invasion, n 0.018
   Yes 20 17
   No 25 55
Lymph node metastasis, n 0.110
   Yes 23 26
   No 22 46
CEA (ng/ml) 0.701
   ≤ 12 41 67
   > 12 4 5
CA19-9 (U/ml) 0.832
   ≤ 282 29 45
   > 282 16 27
LAMA3 expression, n 0.005
   High 30 29
   Low 15 43
CEA = carcinoembryonic antigen; CA19-9 = carbohydrate antigen 19-9; LAMA3 = laminin 
subunit alpha 3
found that LAMA3 expression levels correlated 
with the histological grade of tumors in patients 
with PDAC. However, our analysis of clinical data 
revealed that high LAMA3 expression was associ-
ated with larger tumor size, advanced TNM stage, 
and liver metastasis. This discrepancy may be due 
to the inherent bias from our relatively small sam-
ple size. Jun et al. demonstrated that overexpres-
sion of the α3, β3, and γ2 chains of LM-332 might 
play a crucial role in the progression and progno-
sis of PDAC.15 Based on these findings, we evalu-
ated the prognostic value of LAMA3 expression 
in patients with PDAC. Using the Kaplan–Meier 
Plotter dataset, which incorporates data from the 
GEO, European Genome-phenome Archive, and 
TCGA databases, we found that high LAMA3 
expression was strongly associated with worse 
OS and RFS in patients with PDAC. Additionally, 
we followed up 117 patients with PDAC and ob-
served that high LAMA3 expression in PDAC was 
correlated with poor OS, indicating an overall 
poor prognosis. Univariate and multivariate Cox 
regression analyses further demonstrated that 
LAMA3 was an independent predictive factor for 
mortality in patients with PDAC. In conclusion, 
these results strongly support that the expression 
of LAMA3 can serve as a robust prognostic bio-
marker of PDAC.
Metastasis, the primary cause of cancer-related 
mortality, continues to be an area of limited un-
derstanding regarding its cellular and molecular 
mechanisms.16 Various studies have implicated 
LAMA3 in different mechanisms of metastasis. 
Shu et al. demonstrated that the overexpression of 
LINC00936 hindered ovarian cancer progression 
by competitively binding to miR-221-3p and mod-
ulating LAMA3 expression.7 Moreover, Xu et al. re-
ported that LINC00628 could obstruct cell prolif-
eration, invasion, migration, and apoptosis while 
reducing drug resistance in lung adenocarcinoma 
cells by downregulating the methylation of the 
LAMA3 promoter8. Kinoshita et al. demonstrated 
that miRNA-218 modulated the focal adhesion 
pathway, thereby impeding tumor cell invasion 
and metastasis.9 The liver, which serves as the pri-
mary blood-borne drainage site for related organs, 
such as the portal vein system, colon, and pan-
creas, is crucial for distant metastasis in patients 
with PDAC.17 To gain a deeper understanding of 
the correlation between LAMA3 and liver metas-
tasis in PDAC, we examined independent risk fac-
tors associated with liver metastasis. Univariate 
analysis revealed that histological grade, TNM 
stage, vascular invasion, and high LAMA3 expres-
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Xing Y et al./ Laminin subunit alpha 3 in pancreatic ductal adenocarcinoma 241
sion were significantly associated with liver me-
tastasis. Multivariate logistic regression analyses 
demonstrated that LAMA3 and histological grade 
were independent predictive factors for liver me-
tastasis in patients with PDAC. These findings in-
dicate that poor differentiation and high LAMA3 
expression are correlated with an increased risk of 
metastasis.
This study has both strengths and limitations, 
necessitating further investigations to confirm 
and expand our findings. One of the strengths of 
this study is the use of data from public databases 
combined with bioinformatics analysis. This ap-
proach allows the utilization of large amounts of 
data, thus increasing the reliability and statistical 
power of our findings. We complemented this da-
tabase analysis with clinical data to verify our re-
sults by adding another validation layer. However, 
this study has some limitations. This study was 
entirely based on data from public databases; al-
though we used clinical data to confirm our find-
ings, future studies with larger sample sizes and 
varied population groups are required to further 
validate our results. We evaluated LAMA3 expres-
sion in PDAC tissues using immunohistochemis-
try, which, although a common and reliable tech-
nique, only provides a snapshot of LAMA3 expres-
sion and does not provide functional information. 
Therefore, additional functional experiments are 
required to better understand the role of LAMA3 
in PDAC. Finally, our study did not fully explore 
the mechanism by which LAMA3 promotes liver 
metastasis in PDAC. Understanding these mecha-
nisms requires a series of in-depth molecular and 
cellular biology studies involving in vitro and in 
vivo models. By identifying and understanding 
the precise mechanisms involved, new potential 
therapeutic targets for PDAC can be identified.
This retrospective analysis suggests that 
LAMA3 may serve as a potential biomarker for 
predicting the prognosis of patients with PDAC. 
The increase in LAMA3 expression in PDAC tis-
sues and its association with liver metastasis fur-
ther underscore its potential role in disease pro-
gression. While these findings provide important 
insights, they also highlight the need for further 
studies. Understanding the specific mechanisms 
by which LAMA3 contributes to PDAC progres-
sion and liver metastasis may help uncover new 
therapeutic targets, potentially leading to more 
personalized treatment strategies.
TABLE 3. Univariate and multivariate Cox proportional hazard regression analyses of overall survival
Variables
Univariate analysis Multivariate analysis
HR (95% CI) P-value HR 95% CI) P-value
Age 1.009 (0.985–1.035) 0.459 1.009 (0.983–1.035) 0.512
Sex 1.346 (0.850–2.131) 0.205 0.696 (0.435–1.112) 0.696
Tumor location (head vs. body and tail) 0.895 (0.573–1.399) 0.628 0.780 (0.474–1.283) 0.327
Tumor size
 ≤ 2 cm vs. > 4 cm 0.543 (0.186–1.583) 0.263 0.626 (0.191–2.054) 0.440
  > 2 cm and ≤ 4 cm vs. > 4 cm 0.607 (0.373–0.987) 0.044 0.637 (0.337–1.204) 0.165
Histological grade (G1 vs. G2–3) 1.263 (0.778–2.049) 0.345 1.378 (0.797–2.383) 0.251
TNM stage (I–II vs. III–IV) 0.374 (0.227–0.615) < 0.001 1.505 (0.855–2.647) 0.157
Perineural invasion (yes vs. no) 0.803 (0.464–1.389) 0.432 1.158 (0.611–2.196) 0.652
Vascular invasion (yes vs. no) 0.693 (0.442–1.085) 0.109 0.912 (0.529–1.572) 0.741
Lymph node metastasis (yes vs. no) 0.796 (0.513–1.235) 0.308 0.883 (0.548–1.423) 0.609
Liver metastasis (yes vs. no) 0.364 (0.231–0.574) < 0.001 2.284 (1.426–3.657) 0.001
CEA (≤12 vs. >12) 0.512 (0.256–1.026) 0.059 1.622 (0.788–3.340) 0.189
CA19-9(≤282 vs. >282) 0.969 (0.613–1.533) 0.894 0.963 (0.560–1.655) 0.891
LAMA3 (low vs. high) 0.407 (0.259–0.641) < 0.001 2.016 (1.257–3.234) 0.004
CA19-9 = carbohydrate antigen 19-9; CEA = carcinoembryonic antigen; CI = confidence interval; HR = hazard ratio; LAMA3 = laminin subunit 
alpha 3
Radiol Oncol 2024; 58(2): 234-242.
Xing Y et al./ Laminin subunit alpha 3 in pancreatic ductal adenocarcinoma242
Increased LAMA3 expression is associated with 
poor prognosis and liver metastasis in patients 
with PDAC. Our results indicate that LAMA3 can 
be a novel predictor of poor prognosis in patients 
with PDAC and liver metastasis, and LAMA3 may 
be a promising candidate for targeted therapy for 
PDAC liver metastasis.
Acknowledgements
Funding statement
This study was supported by the Natural 
Science Foundation of Shandong Province (No. 
ZR2020MH059)
Author contributions
All authors contributed to the study conception 
and design. Material preparation, data and speci-
men collection and analysis were performed by 
YyX and GQ. The first draft of the manuscript was 
written by YyX. The institutional review board 
approval and the direction of experiments by XJ. 
YpJ is the guarantor of the study and revised the 
manuscript. All authors commented on previous 
versions of the manuscript. All authors read and 
approved the final manuscript.
Data availability statement
The data that support the findings of this study are 
openly available from The Cancer Genome Atlas 
database (https://portal.gdc.cancer.gov/) and the 
Gene Expression Omnibus database (https://www.
ncbi.nlm.nih.gov/geo). The clinical data that sup-
port the findings of this study are available from 
the author, upon reasonable request.
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