高表达PRELID1促进胃癌细胞上皮间质转化并与不良预后相关

doi:10.12122/j.issn.1673-4254.2025.07.21

摘要/Abstract

摘要：

目的探究含PRELI域1（PRELID1）在胃癌组织中的表达水平及对患者预后和癌细胞上皮间质转化的影响与可能的调控机制。方法纳入115例我院在2018年2月~2023年3月接受胃癌根治术的患者，探究PRELID1在胃癌组织中的表达水平与肿瘤进展和预后的关系。使用慢病毒转染干预MGC803和AGS胃癌细胞PRELID1的表达水平，将细胞分为3组：对照组（NC）、PRELID1过表达组（LV组）和干扰组（siRNA组），分析其对细胞迁移、侵袭和上皮间质转化的影响，以及相关的分子机制。结果免疫组化结果显示，PRELID1在胃癌组织中的表达水平高于癌旁组织（P<0.05），且和患者的恶性进展临床参数［CEA≥5 ng/mL（P=0.007）、CA199≥37 U/mL（P=0.007）、G_3~4（P=0.001）、T_3~4（P=0.001）和N_2~3期（P=0.020）］呈正相关。依据单因素分析和Cox多因素分析的结果提示PRELID1高表达是影响胃癌5年生存期的独立危险因素（P=0.001）。CCK-8实验结果显示，过表达PRELID1促进胃癌细胞的增殖（P<0.05），而干扰PRELID1则抑制（P<0.05）；Transwell迁移和侵袭实验数据提示，过表达PRELID1上调胃癌细胞的迁移与侵袭数量（P<0.05），而干扰PRELID1则结果相反（P<0.05）；免疫印迹检测发现，过表达PRELID1增强胃癌细胞中基质金属蛋白酶（MMP）的表达（P<0.05），而干扰PRELID1则反之（P<0.05）。过表达PRELID1上调胃癌细胞中N-cadherin和vimentin的表达（P<0.05），以及下调E-cadherin的表达（P<0.05）；而干扰PRELID1则结果相反（P<0.05）。过表达胃癌细胞中PRELID1增加了p-PI3K、p-AKT和p-mTOR的表达（P<0.05），而干扰则抑制（P<0.05）；相对干扰PRELID1组的细胞，PI3K/AKT通路激活剂（740 Y-P）增加了细胞迁移和侵袭的数量（P<0.05），以及上调了N-cadherin和vimentin的表达与下调E-cadherin的表达（P<0.05）。结论 PRELID1高表达于胃癌中且影响患者预后，其可能通过激活PI3K/AKT/mTOR通路，进而增强胃癌细胞的迁移和侵袭及上皮间质转化。

关键词: 胃癌, PRELI域1, 预后, 上皮间质转化, PI3K/AKT/mTOR

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

吴璇, 方家敏, 韩玮玮, 陈琳, 孙菁, 金齐力. 高表达PRELID1促进胃癌细胞上皮间质转化并与不良预后相关[J]. 南方医科大学学报, 2025, 45(7): 1535-1542.

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.

图/表 8

图1 胃癌组织中的PRELID1表达情况

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.

表1 PRELID1在胃癌中表达水平与患者临床病理参数间的关系

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

图2 PRELID1表达水平对患者术后5年生存期的影响

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.

表2 影响115例胃癌根治术后5年生存期的单因素及多因素分析

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

图3 调控胃癌细胞PRELID1的表达对增殖、迁移和侵袭的影响

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.

图4 调控PRELID1的表达对胃癌细胞上皮间质转化的影响

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.

图5 评估PI3K/AKT/mTOR通路是否参与PRELID1对胃癌细胞上皮间质转化的调控作用

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.

图6 在体验证调控胃癌细胞PRELID1对移植瘤生长的影响

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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