Introduction
Materials and methods
Patients and samples
Extraction and tryptic digestion of protein
Quantitative proteomic analysis
Targeted proteomics analysis
Retrospective immunohistochemical analysis
Cell culture
Transfection of small interfering RNA and evaluation biological behaviors of cervical cancer cells
Bioinformatics and statistics
Results
Screening of candidate proteins using untargeted quantitative proteomic
Validation of candidate proteins and identification COPA as a potential biomarker using targeted proteomics
Protein Accession | Gene name | SCC/H Ratio | SCC/H p value | AC/H Ratio | AC/H p value | AC/SCC Ratio | AC/SCC p value |
---|---|---|---|---|---|---|---|
P53621 | COPA | 2.07 | 4.00E−03 | 2.80 | 2.53E−04 | 1.35 | 2.95E−02 |
Q7Z3K3 | POGZ | 4.74 | 1.46E−04 | 6.43 | 4.98E−05 | 1.36 | 5.30E−02 |
Q8N2K0 | ABHD12 | 2.86 | 2.12E−06 | 3.93 | 6.60E−03 | 1.37 | 1.11E−01 |
P04275 | VWF | 0.72 | 1.30E−01 | 0.75 | 2.07E−01 | 1.04 | 7.68E−01 |
Q6YHK3 | CD109 | 1.08 | 6.71E−01 | 0.47 | 1.05E−03 | 0.44 | 3.49E−05 |
P52209 | PGD | 5.00 | 1.83E−03 | 3.00 | 1.12E−02 | 0.60 | 2.74E−02 |
P27482 | CALML3 | 6.85 | 4.03E−03 | 0.93 | 9.36E−01 | 0.14 | 5.31E−05 |
P02671 | FGA | 0.35 | 1.99E−02 | 0.54 | 1.06E−01 | 1.53 | 2.61E−02 |
P02679 | FGG | 0.42 | 1.23E−02 | 0.61 | 8.61E−02 | 1.45 | 1.83E−02 |
P00568 | AK1 | 0.90 | 5.27E−01 | 1.23 | 3.65E−01 | 1.37 | 5.33E−02 |
Q9UIJ7 | AK3 | 0.96 | 7.95E−01 | 0.95 | 8.07E−01 | 0.99 | 9.55E−01 |
P43034 | PAFAH1B1 | 1.31 | 1.26E−01 | 1.27 | 1.12E−01 | 0.97 | 7.92E−01 |
Q14624 | ITIH4 | 0.48 | 7.14E−05 | 0.56 | 1.23E−03 | 1.16 | 2.92E−01 |
P01860 | IGHG3 | 0.46 | 2.71E−05 | 0.44 | 1.32E−04 | 0.95 | 7.86E−01 |
Q9H223 | EHD4 | 1.22 | 2.67E−01 | 1.33 | 1.35E−01 | 1.09 | 5.04E−01 |
P42765 | ACAA2 | 0.97 | 8.73E−01 | 1.76 | 1.26E−01 | 1.81 | 1.80E−02 |
P02649 | APOE | 0.28 | 5.01E−06 | 0.46 | 6.79E−03 | 1.64 | 6.50E−02 |
Q9BSJ8 | ESYT1 | 1.63 | 4.62E−04 | 1.62 | 7.65E−03 | 0.99 | 9.46E−01 |
O60716 | CTNND1 | 5.21 | 5.00E−04 | 3.34 | 3.27E−04 | 0.64 | 1.79E−02 |
Functional analysis of COPA using bioinformatics
Correlation analysis between COPA expression and clinicopathological characteristics in IHC cohort
Clinicopathological characteristics | Cervical AC (n = 85) | Cervical SCC (n = 55) | ||||||
---|---|---|---|---|---|---|---|---|
Score 0 n (%) | Score 1 n (%) | Score 2 + n (%)3 | P value | Score 0 n (%) | Score 1 n (%) | Score 2 + n (%)3 | p value | |
Stage | ||||||||
I | 1 (1.2) | 7 (8.2) | 50 (58.8) | 0.455 | 2 (3.6) | 18 (32.7) | 27 (49.1) | 0.536 |
II | 0 (0) | 0 (0) | 20 (23.5) | 0 (0) | 1 (1.8) | 5 (9.1) | ||
III | 0 (0) | 0 (0) | 7 (8.2) | 0 (0) | 0 (0) | 2 (3.6) | ||
PI | ||||||||
Yes | 0 (0) | 0 (0) | 2 (2.4) | 1 | 0 (0) | 1 (1.8) | 1 (1.8) | 1 |
No | 1 (1.2) | 7 (8.2) | 75 (88.2) | 2 (3.6) | 18 (32.7) | 33 (60.0) | ||
LNM | ||||||||
Yes | 0 (0) | 1 (1.2) | 10 (11.8) | 1 | 1 (1.8) | 2 (3.6) | 6 (10.9) | 0.331 |
No | 1 (1.2) | 6 (7.1) | 67 (78.8) | 1 (1.8) | 17 (30.9) | 28 (50.9) | ||
TS | ||||||||
≥ 2 cm | 1 (1.2) | 1 (1.2) | 46 (54.1) | 0.040 | 2 (3.6) | 14 (25.5) | 24 (43.6) | 1 |
< 2 cm | 0 (0) | 6 (7.1) | 31 (36.5) | 0 (0) | 5 (9.1) | 10 (18.1) | ||
DSI | ||||||||
Yes | 0 (0) | 3 (3.5) | 43 (50.6) | 0.558 | 2 (3.6) | 14 (25.5) | 23 (41.8) | 0.880 |
No | 1 (1.2) | 4 (4.7) | 34 (40.0) | 0 (0) | 5 (9.1) | 11 (20.0) | ||
LVSI | ||||||||
Yes | 0 (0) | 1 (1.2) | 14 (16.5) | 1 | 1 (1.8) | 6 (10.9) | 16 (29.1) | 0.551 |
No | 1 (1.2) | 6 (7.1) | 63 (74.1) | 1 (1.8) | 13 (23.6) | 18 (32.7) |
Histology
Age
Stage
Parametrial invasion
Lymph node metastasis
Tumor size
Deep stromal invasion
Lymphatic vascular space involvement
Validation of COPA staining can serve as an independent prognostic factor for cervical cancer
Variables | Univariate analysis | Multivariate analysis | ||||
---|---|---|---|---|---|---|
HR | 95% CI | p value | HR | 95% CI | p value | |
COPA | 10.773 | 1.479–78.497 | 0.019* | 8.946 | 1.218–65.714 | 0.031* |
Stage | 5.427 | 2.546–11.565 | < 0.001* | 3.624 | 1.459–8.999 | 0.006* |
LNM | 2.304 | 1.146–4.629 | 0.019* | 1.354 | 0.584–3.139 | 0.48 |
TS | 1.243 | 0.637–2.426 | 0.523 | |||
DSI | 1.455 | 0.737–2.874 | 0.28 | |||
PI | 0.97 | 0.133–7.076 | 0.976 | |||
LVSI | 1.501 | 0.780–2.888 | 0.224 |