High PRELID1 expression promotes epithelial-mesenchymal transition in gastric cancer cells and is associated with poor prognosis

doi:10.12122/j.issn.1673-4254.2025.07.21

Abstract

Abstract:

Objective To investigate the correlation of PRELID1 with gastric cancer (GC) progression, prognosis, and epithelial-mesenchymal transition (EMT) and the underlying mechanisms. Methods We analyzed the data of 115 patients undergoing radical gastrectomy for GC in our hospital between February, 2018 and March, 2023 to explore the correlation of PRELID1 expression level in GC tissues with tumor progression and patient prognosis. In cultured GC cells, the effects of lentivirus-mediated overexpression or interference of PRELID1 were observed on cell migration, invasion and EMT. Results Immunohistochemical staining revealed significantly higher PRELID1 expression in GC tissues (P<0.001), whose expression level was positively correlated with CEA ≥5 ng/mL (P=0.007), CA199 ≥37 U/mL (P=0.007), G_3-4 stages (P=0.001), T_3-4 stages (P=0.001), and N_2-3 stages (P=0.020). Univariate and Cox multifactorial analysis showed that high PRELID1 level was an independent risk factor affecting 5-year survival of GC patients (P=0.001). In cultured GC cells, PRELID1 overexpression obviously promoted cell proliferation, migration, invasion, and the expressions of MMP2 and MMP9, and interference of PRELID1 produced the opposite changes. PRELID1 overexpression also increased the expressions of N-cadherin and vimentin and reduced the expression of E-cadherin. Mechanistic analyses showed that up-regulation of PRELID1 increased the expression of p-PI3K, p-AKT, and p-mTOR in GC cells, whereas its interference caused the opposite changes; the application of 740 Y-P, a PI3K/AKT pathway activator, significantly enhanced the migration, invasion, and EMT of GC cells with PRELID1 knockdown. Conclusion PRELID1 is highly expressed in GC and affects prognosis of the patients, and its high expression promotes migration, invasion and epithelial mesenchymal transition of GC cells possibly by activating the PI3K/AKT/mTOR pathway.

Key words: gastric cancer, PRELID1, prognosis, epithelial mesenchymal transition, PI3K/AKT/mTOR

Xuan WU, Jiamin FANG, Weiwei HAN, Lin CHEN, Jing SUN, Qili JIN. High PRELID1 expression promotes epithelial-mesenchymal transition in gastric cancer cells and is associated with poor prognosis[J]. Journal of Southern Medical University, 2025, 45(7): 1535-1542.

Figures/Tables 8

Fig 1 PRELID1 expression in gastric cancer and adjacent tissues. A: mRNA level of PRELID1 detected by qRT-PCR (n=10). B: Immunohistochemistry of PRELID1 expression in gastric cancer and adjacent tissues. C: Relative integrated optical density value of PRELID1 in gastric cancer and adjacent tissues (Mean±SD, n=52). *P<0.05 vs Adjacent.

Tab.1 Correlation of PRELID1 expression levels in gastric cancer with clinicopathological parameters of patients

Characteristic	n	PRELID1 expression (n, %)		χ²	P
Characteristic	n	Low expression (n=58)	High expression (n=57)	χ²	P
Gender
Female	55	32 (58.2%)	23 (41.8%)	2.531	0.112
Male	60	26 (43.3%)	34 (56.7%)	2.531	0.112
Age(year)
<60	45	24 (53.3%)	21 (46.7%)	0.248	0.618
≥60	70	34 (48.6%)	36 (51.4%)	0.248	0.618
CEA (ng/mL)
<5	61	38 (62.3%)	23 (37.7%)	7.31	0.007
≥5	54	20 (37.0%)	34 (63.0%)	7.31	0.007
CA199 (U/mL)
<37	59	37 (62.7%)	22 (37.3%)	7.305	0.007
≥37	56	21 (37.5%)	35 (62.5%)	7.305	0.007
Tumor size (cm)
<5	57	31 (54.4%)	26 (45.6%)	0.706	0.401
≥5	58	27 (46.6%)	31 (53.4%)	0.706	0.401
Pathological type
Adenocarcinoma	93	49 (52.7%)	44 (47.3%)	0.987	0.32
Other	22	9 (40.9%)	13 (59.1%)	0.987	0.32
Pathological grading
G1-G2	62	40 (64.5%)	22 (35.5%)	10.671	0.001
G3-G4	53	18 (34.0%)	35 (66.0%)	10.671	0.001
T Stage
T1-T2	57	38 (66.7%)	19 (33.3%)	11.912	0.001
T3-T4	58	20 (34.5%)	38 (65.5%)	11.912	0.001
N Stage
N0-N1	63	38 (60.3%)	25 (39.7%)	5.443	0.02
N2-N3	52	20 (38.5%)	32 (61.5%)	5.443	0.02

Fig.2 Effect of PRELID1 expression level on 5-year postoperative survival of gastric cancer patients. A: Kaplan-Meier survival curve analysis. B: ROC analysis.

Tab.2 Univariate and multivariate analyses of factors affecting 5-year survival of the patients after radical gastrectomy (n=115)

Factors	Univariate analysis		Multivariate analysis
Factors	Log-rank χ²	P	HR	95% CI	P
Gender (male vs female)	2.758	0.097
Age (<60 year vs ≥60 year)	0.063	0.801
PRELID1 expression (high vs low)	27.093	<0.001	2.776	1.518-5.075	0.001
CEA level (<5 ng/mL vs ≥5 ng/mL)	32.566	<0.001	2.297	1.188-4.440	0.013
CA199 level (<37 U/mLvs ≥37 U/mL)	42.932	<0.001	2.570	1.254-5.268	0.010
Cancer cell type (adenocarcinoma vs other)	2.490	0.115
Pathological grading (G1-G2 vs G3-G4)	30.733	<0.001	2.512	1.377-4.584	0.003
Tumor size (<5 cm vs ≥5 cm)	1.250	0.264
T stage (T1~T2 vs T3~T4)	47.055	<0.001	2.959	1.394-6.281	0.005
N stage (N0~N1 vs N2~N3)	44.963	<0.001	2.375	1.201-4.697	0.013

Fig.3 Effects of PRELID1 overexpression and knockdown on proliferation, migration and invasion of gastric cancer cells. A: Western blotting for verifying expression of PRELID1 in gastric cancer cells after transfection. B: CCK-8 assay for analyzing the effect of PRELID1 overexpression and knockdown on proliferation of gastric cancer cells. C-E: Transwell migration and invasion assay for evaluating the effect of PRELID1 overexpression and knockdown on migration and invasion of gastric cancer cells. F: Western blotting for detecting expressions of MMP-2 and MMP-9. *P<0.05 vs NC; #P<0.05 vs NC.

Fig.4 Effect of PRELID1 overexpression and knockdown on epithelial-mesenchymal transition in gastric cancer cells. A: qRT-PCR analysis of the effect of PRELID1 overexpression and knockdown on mRNA expression of CDH1, CDH2 and vimentin in gastric cancer cells. B: Western blotting for detecting E-cadherin, N-cadherin and vimentin expressions in the transfected cells. *P<0.05 vs NC, #P<0.05 vs NC.

Fig.5 Assessment of the role of the PI3K/AKT/mTOR pathway in regulating epithelial-mesenchymal transition in gastric cancer cells with PRELID1 overexpression and knockdown. A: Western blotting for detecting the expressions of p-PI3K, p-AKT and p-mTOR in the treated gastric cancer cells. B, C: Transwell migration and invasion assay to assess the effect of PI3K/AKT pathway activator (740 Y-P) on the migration and invasion of gastric cancer cells. D: Western blotting for detecting MMP-2, MMP-9, E-cadherin, N-cadherin and vimentin expression levels in the treated cells. E, F: Transwell migration and invasion assay for evaluating the effects of PI3K/AKT pathway inhibitor (LY294002) on gastric cancer cell migration and invasion. *P<0.05 vs siRNA of LV.

Fig.6 Effect of PRELID1 overexpression and knockdown on growth of gastric cancer cell xenografts in nude mice. A: Gross observation of the dissected tumors in the 3 groups. B: Comparison of the tumor volume among the 3 groups. C: Comparison of tumor weight among the 3 groups. *P<0.05 vs NC, #P<0.05 vs NC.

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