Screening of diagnostic and prognostic markers of pancreatic cancer based on multi-omics analysis of serum and tissue exosomes

doi:10.3877/cma.j.issn.2095-3232.2025.06.022

Abstract

Abstract:

Objective

To screen early diagnostic and prognostic markers of pancreatic ductal adenocarcinoma (PDAC) based on multi-omics analysis of serum and tissue exosomes.

Methods

The samples were collected from 8 PDAC patients undergoing surgery and 3 healthy donors admitted to Zhongnan Hospital of Wuhan University in 2020. Exosomes were isolated from tissues and serum of healthy donors and PDAC patients. The particle size and protein content were identified by electron microscope and molecular markers. Subsequently, PDAC-specific exosomal microRNA (miRNA) was identified by miRNA transcriptomics. The verified samples were obtained from 22 PDAC patients and 20 healthy donors to further verify the identified miRNAs. In the final analysis, the diagnostic ability of these miRNAs for PDAC was evaluated by the area under the ROC curve (AUC) and Brier score. Kaplan-Meier method and Log-rank test were used for survival analysis.

Results

5 miRNAs with low expression levels (hsa-miR-142-3p, hsa-miR-199b-3p, hsa-miR-221-3p, hsa-miR-222-3p and hsa-miR-584-5p) and 1 with high expression level were identified and verified. Two miRNAs with the highest expression levels were miR142-3p and miR148a-3p. Compared with adjacent tissues and healthy donor sera, these two miRNAs were further verified in tissue-derived extracellular vesicles (Ti-EVs) and serum-derived extracellular vesicles (Se-EVs) of PDAC patients. Kaplan-Meier curve showed that the down-regulated expression levels of miR142-3p and miR148a-3p were significantly correlated with poor prognosis of overall survival (OS) and recurrence-free survival (RFS) of PDAC patients (χ²=36.314, 5.218 and 7.047, 4.924; all P<0.05). The prediction error curve indicated that the combination of miR142-3p, miR148a-3p and CA19-9 yielded the smallest prediction error. ROC analysis revealed that the AUC based on the combination of miR142-3p and miR148a-3p combined with CA19-9 model was 0.747, larger than that of the model using CA19-9 alone.

Conclusions

The exosomes of pancreatic cancer, miRNA-142-3p and miRNA-148a-3p, play an anti-oncogene role in PDAC. PDAC patients with low expression of miRNA-142-3p or miRNA-148a-3p achieve poor prognosis, which are novel potential diagnostic and prognostic markers for PDAC.

Key words: Pancreatic ductal adenocarcinoma (PDAC), microRNA (miRNA), Exosome, Diagnosis, Prognosis

Gang Wu, Ranxing Yan, Xin Yan, Jing Yan, Yueming He, Qian Zhu. Screening of diagnostic and prognostic markers of pancreatic cancer based on multi-omics analysis of serum and tissue exosomes[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2025, 14(06): 962-972.

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References 32

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样本	年龄(岁)	性别	胆囊炎	黄疸	消瘦	减轻黄疸	CA19-9(kU/L)	肿瘤大小(cm)	TNM 分期	包膜浸润	脂肪浸润	血管浸润	输血	术后出血
T1-N1	75	男	无	无	无	无	>1 000	4×3×3	T2N0M0	有	有	无	有	无
T2-N2	60	男	无	无	无	无	1 000	3×2×2	T2N0M1	有	有	有	有	无
T3-N3	60	男	无	无	无	无	40.05	4×4×3	T2N2M0	有	有	有	无	有
T4-N4	58	男	无	无	无	无	1 081	4×4×4	T2N0M0	有	无	有	无	无
T5-N5	69	男	有	有	有	PTCD	>1 000	3×2×1	T2N2M0	无	有	有	有	无
T6-N6	63	女	无	无	无	无	>1 000	3×2×1	T2N1M0	无	无	有	无	无
T7-N7	68	男	无	有	有	PTCD	920.10	3×2×1	T2N0M0	无	无	有	无	有
T8-N8	49	女	有	有	无	PTCD	30.53	2×2×1	T1cN0M0	无	无	无	无	无

样本	年龄(岁)	性别	胆囊炎	黄疸	消瘦	减轻黄疸	CA19-9(kU/L)	肿瘤大小(cm)	TNM 分期	包膜浸润	脂肪浸润	血管浸润	输血	术后出血
T1-N1	75	男	无	无	无	无	>1 000	4×3×3	T2N0M0	有	有	无	有	无
T2-N2	60	男	无	无	无	无	1 000	3×2×2	T2N0M1	有	有	有	有	无
T3-N3	60	男	无	无	无	无	40.05	4×4×3	T2N2M0	有	有	有	无	有
T4-N4	58	男	无	无	无	无	1 081	4×4×4	T2N0M0	有	无	有	无	无
T5-N5	69	男	有	有	有	PTCD	>1 000	3×2×1	T2N2M0	无	有	有	有	无
T6-N6	63	女	无	无	无	无	>1 000	3×2×1	T2N1M0	无	无	有	无	无
T7-N7	68	男	无	有	有	PTCD	920.10	3×2×1	T2N0M0	无	无	有	无	有
T8-N8	49	女	有	有	无	PTCD	30.53	2×2×1	T1cN0M0	无	无	无	无	无

序号	G1	G2	G3	G4
1	hsa-let-7c-5p	hsa-let-7c-5p	hsa-miR-16-5p	hsa-miR-16-5p
2	hsa-let-7b-5p	hsa-let-7b-5p	hsa-let-7a-5p	hsa-let-7c-5p
3	hsa-let-7a-5p	hsa-let-7a-5p	hsa-let-7f-5p	hsa-let-7a-5p
4	hsa-let-7f-5p	hsa-let-7f-5p	hsa-let-7c-5p	hsa-let-7f-5p
5	hsa-let-7e-5p	hsa-miR-125b-5p	hsa-miR-126-3p	hsa-miR-126-3p
6	hsa-miR-148a-3p	hsa-miR-125a-5p	hsa-let-7e-5p	hsa-let-7e-5p
7	hsa-miR-375-3p	hsa-miR-16-5p	hsa-miR-142-3p	hsa-miR-122-5p
8	hsa-miR-125b-5p	hsa-let-7e-5p	hsa-let-7b-5p	hsa-let-7b-5p
9	hsa-miR-125a-5p	hsa-miR-21-5p	hsa-miR-223-3p	hsa-miR-486-5p
10	hsa-miR-16-5p	hsa-miR-199b-3p	hsa-let-7i-5p	hsa-let-7i-5p
11	hsa-miR-21-5p	hsa-let-7i-5p	hsa-miR-26a-5p	hsa-miR-92a-3p
12	hsa-miR-30a-5p	hsa-miR-148a-3p	hsa-miR-26b-5p	hsa-miR-142-3p
13	hsa-miR-30d-5p	hsa-miR-30a-5p	hsa-miR-30a-5p	hsa-miR-223-3p
14	hsa-miR-199b-3p	hsa-miR-143-3p	hsa-miR-486-5p	hsa-miR-26b-5p
15	hsa-miR-200c-3p	hsa-miR-26a-5p	hsa-miR-21-5p	hsa-miR-30a-5p
16	hsa-let-7i-p	hsa-miR-30d-5p	hsa-miR-92a-3p	hsa-miR-26a-5p
17	hsa-miR-26a-5p	hsa-miR-200c-3p	hsa-miR-103a-3p	hsa-miR-21-5p
18	unconservative_21_1433793	hsa-miR-7-5p	hsa-miR-107	hsa-miR-103a-3p
19	hsa-miR-7-5p	hsa-miR-26b-5p	hsa-miR-30d-5p	hsa-miR-107
20	hsa-miR-26b-5p	hsa-miR-375-3p	hsa-miR-146a-5p	hsa-miR-93-5p

序号	G1	G2	G3	G4
1	hsa-let-7c-5p	hsa-let-7c-5p	hsa-miR-16-5p	hsa-miR-16-5p
2	hsa-let-7b-5p	hsa-let-7b-5p	hsa-let-7a-5p	hsa-let-7c-5p
3	hsa-let-7a-5p	hsa-let-7a-5p	hsa-let-7f-5p	hsa-let-7a-5p
4	hsa-let-7f-5p	hsa-let-7f-5p	hsa-let-7c-5p	hsa-let-7f-5p
5	hsa-let-7e-5p	hsa-miR-125b-5p	hsa-miR-126-3p	hsa-miR-126-3p
6	hsa-miR-148a-3p	hsa-miR-125a-5p	hsa-let-7e-5p	hsa-let-7e-5p
7	hsa-miR-375-3p	hsa-miR-16-5p	hsa-miR-142-3p	hsa-miR-122-5p
8	hsa-miR-125b-5p	hsa-let-7e-5p	hsa-let-7b-5p	hsa-let-7b-5p
9	hsa-miR-125a-5p	hsa-miR-21-5p	hsa-miR-223-3p	hsa-miR-486-5p
10	hsa-miR-16-5p	hsa-miR-199b-3p	hsa-let-7i-5p	hsa-let-7i-5p
11	hsa-miR-21-5p	hsa-let-7i-5p	hsa-miR-26a-5p	hsa-miR-92a-3p
12	hsa-miR-30a-5p	hsa-miR-148a-3p	hsa-miR-26b-5p	hsa-miR-142-3p
13	hsa-miR-30d-5p	hsa-miR-30a-5p	hsa-miR-30a-5p	hsa-miR-223-3p
14	hsa-miR-199b-3p	hsa-miR-143-3p	hsa-miR-486-5p	hsa-miR-26b-5p
15	hsa-miR-200c-3p	hsa-miR-26a-5p	hsa-miR-21-5p	hsa-miR-30a-5p
16	hsa-let-7i-p	hsa-miR-30d-5p	hsa-miR-92a-3p	hsa-miR-26a-5p
17	hsa-miR-26a-5p	hsa-miR-200c-3p	hsa-miR-103a-3p	hsa-miR-21-5p
18	unconservative_21_1433793	hsa-miR-7-5p	hsa-miR-107	hsa-miR-103a-3p
19	hsa-miR-7-5p	hsa-miR-26b-5p	hsa-miR-30d-5p	hsa-miR-107
20	hsa-miR-26b-5p	hsa-miR-375-3p	hsa-miR-146a-5p	hsa-miR-93-5p

对比组	COG	GO	KEGG	KOG	NR	Pfam	SWISS-PROT	eggNOG
G1比G2	4 979	14 831	15 293	10 800	15 785	14 839	15 734	15 448
G2比G4	5 918	17 525	18 118	12 618	18 751	17 554	18 661	18 317
G3比G4	3 727	11 181	11 555	8 130	11 922	11 159	11 872	11 684

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