Detection of p53, PGP9.5, SOX2, GAGE7, GBU4-5 and MAGE A1 protein levels in lung cancer tissues and peripheral blood and their clinical value

doi:10.19401/j.cnki.1007-3639.2023.01.004

Abstract

Abstract:

Background and purpose: The pathogenesis of lung cancer is diverse, however once confirmed, the best time for treatment is often missed. In this paper, the correlation between tumor suppressor gene (p53), protein gene product (PGP9.5), transcription factor (SOX2), tumor-associated gene protein (GAGE7), helicase (GBU4-5) and melanoma antigen (MAGE A1) protein levels in lung cancer tissues and peripheral blood of patients was discussed, and its application value in the diagnosis and treatment of lung cancer was analyzed. Methods: A total of 100 lung cancer patients in Fudan University Shanghai Cancer Center from May 2018 to May 2019 were enrolled, including 25 cases of clinical TNM stage Ⅰ, 45 cases of stage Ⅱ, 30 cases of stage Ⅲa, 80 cases of non-small cell lung cancer and 20 cases of small cell lung cancer. Following surgical removal of intact tumor tissues and cancer-adjacent tissues, immunohistochemical staining was used to detect positive rates of above-mentioned 6 proteins, real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) was used to detect their gene expression levels, and enzyme-linked immunosorbent assay (ELISA) was performed to detect antibody positive rates in peripheral serum. Results: The positive rates and gene expression levels of p53, PGP9.5, SOX2, GAGE7, GBU4-5 and MAGE A1 in tumor tissues were significantly higher compared with adjacent tissues (P<0.05). Correlation analysis between p53, PGP9.5, SOX2, GAGE7, GBU4-5 and MAGE A1 proteins in tumor tissues and serum showed that the positive rates and gene expression levels of P53, PGP9.5, SOX2, GAGE7, GBU4-5 and MAGE A1 were correlated with TNM stage and differentiation level (P<0.05). There was no significant difference in gender, age, tumor diameter and pathological type (P>0.05). The protein expression levels of p53, PGP9.5, SOX2, GAGE7, GBU4-5 and MAGE A1 in tumor tissues were consistent with their expression levels in peripheral blood (P>0.05). Conclusion: Positive expressions of p53, PGP9.5, SOX2, GAGE7, GBU4-5 and MAGE A1 protein in lung cancer tissues and peripheral serum were closely related to TNM staging and differentiation.

Key words: Lung cancer, Tumor suppressor gene (p53), Protein gene product (PGP9.5), Transcription factor (SOX2), Tumor-associated gene protein (GAGE7), Helicase (GBU4-5), Melanoma antigen (MAGE A1)

CLC Number:

R734.2

ZOU Chunyuan, XU Xiaofeng, LU Renquan, GUO Lin. Detection of p53, PGP9.5, SOX2, GAGE7, GBU4-5 and MAGE A1 protein levels in lung cancer tissues and peripheral blood and their clinical value[J]. China Oncology, 2023, 33(1): 36-44.

Figures/Tables 6

Tab. 1

Tab. 2

Tab. 3

Tab. 4

Relationship between positive and relative expression levels of protein in tumor tissues and clinicopathological characteristics of tumors [Quantification （n）]"

Item	Case n	p53	PGP9.5	SOX2	GAGE7	GBU4-5	MAGE A1
Gender
Men	60	0.53±0.14 (25)	0.46±0.12 (27)	0.38±0.12 (19)	0.33±0.09 (24)	0.28±0.11 (25)	0.41±0.16 (26)
Women	40	0.51±0.13 (16)	0.49±0.17 (18)	0.39±0.16 (13)	0.36±0.12 (12)	0.29±0.12 (12)	0.43±0.15 (13)
t/χ² value		0.122 (0.028)	0.163 (0.000)	0.155 (0.293)	0.096 (1.042)	0.121 (1.401)	0.113 (1.184)
P value		0.869 (0.868)	0.789 (1.000)	0.825 (0.588)	0.968 (0.307)	0.923 (0.236)	0.869 (0.277)
Age/year
<60.5	42	0.52±0.14 (17)	0.47±0.16 (20)	0.37±0.12 (13)	0.34±0.10 (13)	0.27±0.09 (13)	0.40±0.18 (14)
≥60.5	58	0.54±0.16 (24)	0.49±0.19 (25)	0.40±0.16 (19)	0.36±0.12 (23)	0.30±0.10 (24)	0.43±0.18 (25)
t/χ² value		0.203 (0.008)	0.185 (0.201)	0.252 (0.037)	0.156 (0.801)	0.232 (1.136)	0.257 (0.977)
P value		0.724 (0.928)	0.769 (0.654)	0.689 (0.848)	0.868 (0.371)	0.822 (0.286)	0.786 (0.323)
Tumor diameter/cm
<3.5	46	0.51±0.13 (18)	0.47±0.16 (19)	0.38±0.12 (14)	0.35±0.11 (14)	0.29±0.11 (14)	0.42±0.16 (15)
≥3.5	54	0.55±0.19 (23)	0.49±0.18 (26)	0.40±0.16 (18)	0.36±0.13 (22)	0.31±0.16 (23)	0.43±0.18 (24)
t/χ² value		0.320 (0.123)	0.264 (0.470)	0.358 (0.096)	0.265 (1.145)	0.321 (1.575)	0.367 (1.463)
P value		0.764 (0.726)	0.806 (0.493)	0.702 (0.757)	0.802 (0.285)	0.763 (0.209)	0.712 (0.227)
Pathologic type
SCLC	20	0.52±0.15 (8)	0.46±0.18 (11)	0.37±0.12 (6)	0.33±0.08 (8)	0.28±0.12 (8)	0.41±0.19 (9)
NSCLC	80	0.53±0.16 (33)	0.49±0.18 (34)	0.41±0.16 (26)	0.36±0.12 (28)	0.30±0.13 (29)	0.43±0.21 (30)
t/χ² value		0.524 (0.010)	0.326 (1.010)	0.421 (0.046)	0.326 (0.174)	0.402 (0.097)	0.456 (0.378)
P value		0.523 (0.919)	0.659 (0.315)	0.603 (0.830)	0.721 (0.677)	0.695 (0.756)	0.636 (0.539)
TNM
Ⅰ	25	0.48±0.15 (3)	0.46±0.18 (4)	0.36±0.19 (2)	0.31±0.08 (2)	0.28±0.12 (2)	0.40±0.17 (2)
Ⅱ	45	0.53±0.16 (15)	0.47±0.16 (15)	0.38±0.12 (10)	0.36±0.11 (13)	0.30±0.13 (13)	0.43±0.16 (13)
Ⅲa	30	0.59±0.17 (23)	0.52±0.20 (26)	0.42±0.16 (20)	0.39±0.13 (21)	0.34±0.15 (22)	0.46±0.19 (24)
F/χ² value		5.236 (25.562)	5.524 (32.013)	5.123 (25.163)	4.659 (24.547)	5.003 (27.280)	5.212 (32.231)
P value		0.000 (0.000)	0.000 (0.000)	0.000 (0.000)	0.013 (0.000)	0.007 (0.000)	0.003 (0.000)
Differentiation degree
Low	40	0.60±0.18 (31)	0.53±0.18 (34)	0.43±0.18 (25)	0.41±0.16 (26)	0.33±0.16 (27)	0.47±0.21 (29)
Middle	35	0.55±0.17 (8)	0.48±0.16 (8)	0.38±0.16 (5)	0.37±0.16 (8)	0.29±0.15 (8)	0.44±0.18 (8)
High	25	0.46±0.14 (2)	0.45±0.18 (3)	0.35±0.16 (2)	0.33±0.15 (2)	0.26±0.12 (2)	0.40±0.15 (2)
F/χ² value		5.659 (38.047)	5.857 (43.792)	5.903 (28.765)	5.524 (25.732)	5.968 (27.986)	6.003 (32.802)
P value		0.000 (0.000)	0.000 (0.000)	0.000 (0.000)	0.001 (0.000)	0.000 (0.000)	0.000 (0.000)

Tab. 4

Tab. 5

Relationship between the positive rate and level of p53, PGP9.5, SOX2, GAGE7, GBU4-5, MAGE A1 in serum and the clinicopathological characteristics of tumors [Quantification （n）]"

Item	Case ⁿ	p53	PGP9.5	SOX2	GAGE7	GBU4-5	MAGE A1
Gender
Men	60	6.23±10.84 (22)	1.11±3.12 (25)	5.58±9.14 (17)	3.23±5.09 (24)	1.78±2.71 (22)	2.43±4.76 (24)
Women	40	6.51±9.73 (13)	1.09±3.17 (21)	5.62±8.96 (13)	3.16±5.12 (10)	1.69±2.62 (13)	2.26±4.55 (16)
t/χ² value		0.132 (0.183)	0.031 (1.134)	0.022 (0.499)	0.067 (2.406)	0.165 (0.183)	0.178 (0.000)
P value		0.896 (0.669)	0.975 (0.287)	0.983 (0.480)	0.947 (0.121)	0.869 (0.669)	0.859 (1.000)
Age/year
<60.5	42	6.12±10.14 (14)	1.07±3.16 (19)	5.37±9.12 (11)	3.34±5.11 (11)	1.67±2.59 (11)	2.40±4.68 (13)
≥60.5	58	6.04±10.16 (21)	1.19±3.19 (27)	5.40±8.76 (19)	3.26±5.07 (23)	1.70±2.60 (24)	2.23±4.56 (27)
t/χ² value		0.039 (0.088)	0.186 (0.017)	0.017 (0.500)	0.078 (1.968)	0.057 (2.470)	0.182 (2.470)
P value		0.969 (0.766)	0.953 (0.896)	0.987 (0.479)	0.938 (0.161)	0.955 (0.116)	0.856 (0.116)
Tumor diameter/cm
<3.5	46	6.21±9.73 (15)	1.15±3.13 (21)	5.38±9.15 (11)	3.29±5.16 (13)	1.59±2.61 (13)	2.42±4.46 (15)
≥3.5	54	6.15±10.12 (20)	1.17±2.98 (25)	5.43±8.22 (19)	3.24±5.09 (21)	1.62±2.55 (22)	2.33±4.38 (25)
t/χ² value		0.030 (0.214)	0.033 (0.004)	0.029 (1.503)	0.049 (1.250)	0.058 (1.701)	0.102 (1.939)
P value		0.976 (0.644)	0.974 (0.949)	0.977 (0.220)	0.961 (0.263)	0.954 (0.192)	0.919 (0.164)
Pathologic type
SCLC	20	6.12±9.65 (6)	1.18±3.22 (10)	5.33±9.12 (4)	3.33±5.08 (7)	1.48±2.52 (5)	2.41±4.19 (11)
NSCLC	80	6.03±10.11 (29)	1.22±2.89 (36)	5.41±9.24 (26)	2.26±5.12 (27)	1.60±2.63 (30)	2.32±4.27 (29)
t/χ² value		0.036 (0.275)	0.054 (0.161)	0.035 (1.191)	0.837 (0.011)	0.184 (1.099)	0.085 (2.344)
P value		0.971 (0.600)	0.957 (0.688)	0.972 (0.275)	0.405 (0.916)	0.854 (0.295)	0.933 (0.126)
TNM
Ⅰ	25	5.48±7.15 (1)	0.86±2.16 (5)	5.06±9.15 (1)	2.41±3.08 (1)	1.18±1.82 (2)	2.12±2.13 (3)
Ⅱ	45	6.13±10.16 (13)	1.17±3.11 (13)	5.28±8.92 (11)	3.16±4.11 (11)	1.35±2.13 (11)	2.43±2.06 (11)
Ⅲa	30	6.29±10.08 (21)	1.24±3.22 (28)	5.48±10.11 (18)	3.39±4.13 (23)	1.74±2.55 (22)	2.76±2.24 (26)
F/χ² value		5.965 (13.149)	5.687 (12.842)	5.987 (11.414)	8.655 (16.175)	7.685 (13.239)	6.589 (15.313)
P value		0.001 (0.001)	0.003 (0.002)	0.001 (0.003)	0.001 (0.000)	0.001 (0.001)	0.001 (0.000)
Differentiation degree
Low	40	6.60±4.14 (27)	1.63±2.15 (32)	5.73±5.52 (25)	3.61±3.15 (24)	1.83±2.22 (27)	2.82±2.01 (28)
Middle	35	5.55±4.13 (6)	1.48±2.02 (9)	5.38±5.44 (4)	3.38±4.16 (8)	1.44±1.75 (7)	2.35±2.13 (11)
High	25	5.26±3.18 (2)	1.24±2.08 (5)	5.11±5.36 (1)	3.13±4.12 (2)	1.26±2.11 (1)	2.11±1.85 (1)
F/χ² value		5.258 (14.750)	5.352 (11.137)	5.554 (17.694)	8.524 (10.613)	7.730 (15.682)	7.553 (13.572)
P value		0.005 (0.001)	0.004 (0.004)	0.002 (0.000)	0.001 (0.005)	0.002 (0.000)	0.003 (0.001)

Tab. 5

Fig. 1

References 25

[1]	BADE B C, DELA CRUZ C S. Lung cancer 2020: epidemiology, etiology, and prevention[J]. Clin Chest Med, 2020, 41(1): 1-24. doi: S0272-5231(19)30080-2 pmid: 32008623
[2]	ZHOU Y Y, HÖTI N, AO M H, et al. Expression of p16 and p53 in non-small cell lung cancer: clinicopathological correlation and potential prognostic impact[J]. Biomark Med, 2019, 13(9): 761-771. doi: 10.2217/bmm-2018-0441
[3]	BRAIT M, MALDONADO L, NOORDHUIS M G, et al. Association of promoter methylation of VGF and PGP9.5 with ovarian cancer progression[J]. PLoS One, 2013, 8(9): e70878. doi: 10.1371/journal.pone.0070878
[4]	WANG K X, JI W X, YU Y F, et al. FGFR1-ERK1/2-SOX2 axis promotes cell proliferation, epithelial-mesenchymal transition, and metastasis in FGFR1-amplified lung cancer[J]. Oncogene, 2018, 37(39): 5340-5354. doi: 10.1038/s41388-018-0311-3 pmid: 29858603
[5]	WANG J, SHIVAKUMAR S, BARKER K, et al. Comparative study of autoantibody responses between lung adenocarcinoma and benign pulmonary nodules[J]. J Thorac Oncol, 2016, 11(3): 334-345. doi: 10.1016/j.jtho.2015.11.011 pmid: 26896032
[6]	ORLOVETSKIE N, SERRUYA R, ABBOUD-JARROUS G, et al. Targeted inhibition of WRN helicase, replication stress and cancer[J]. Biochim Biophys Acta Rev Cancer, 2017, 1867(1): 42-48. doi: 10.1016/j.bbcan.2016.11.004
[7]	YI E, CHANG J E, LEEM C, et al. Association of MAGE A1-6 expression with lung cancer progression[J]. J Cancer, 2017, 8(8): 1324-1329. doi: 10.7150/jca.18086 pmid: 28638445
[8]	ZHANG X Z, LIU M, ZHANG X, et al. Autoantibodies to tumor-associated antigens in lung cancer diagnosis[J]. Adv Clin Chem, 2021, 103: 1-45. doi: 10.1016/bs.acc.2020.08.005 pmid: 34229848
[9]	XIAO K J, MA X L, WANG Y C, et al. Diagnostic value of serum tumor-associated autoantibodies in esophageal cancer[J]. Biomark Med, 2021, 15(15): 1333-1343. doi: 10.2217/bmm-2021-0351 pmid: 34541870
[10]	ZHANG R, MA L, LI W Y, et al. Diagnostic value of multiple tumor-associated autoantibodies in lung cancer[J]. Onco Targets Ther, 2019, 12: 457-469. doi: 10.2147/OTT.S187734
[11]	DU Q, YU R F, WANG H, et al. Significance of tumor-associated autoantibodies in the early diagnosis of lung cancer[J]. Clin Respir J, 2018, 12(6): 2020-2028. doi: 10.1111/crj.12769 pmid: 29356386
[12]	CHAPMAN C J, HEALEY G F, MURRAY A, et al. EarlyCDT^®-Lung test: improved clinical utility through additional autoantibody assays[J]. Tumour Biol, 2012, 33(5): 1319-1326. doi: 10.1007/s13277-012-0379-2
[13]	CHAPMAN C J, MURRAY A, MCELVEEN J E, et al. Autoantibodies in lung cancer: possibilities for early detection and subsequent cure[J]. Thorax, 2008, 63(3): 228-233. doi: 10.1136/thx.2007.083592 pmid: 17932110
[14]	INFANTE M, CAVUTO S, LUTMAN F R, et al. A randomized study of lung cancer screening with spiral computed tomography: three-year results from the DANTE trial[J]. Am J Respir Crit Care Med, 2009, 180(5): 445-453. doi: 10.1164/rccm.200901-0076OC
[15]	PARRALES A, IWAKUMA T. Targeting oncogenic mutant p53 for cancer therapy[J]. Front Oncol, 2015, 5: 288. doi: 10.3389/fonc.2015.00288 pmid: 26732534
[16]	QIN J Y, ZENG N, YANG T, et al. Diagnostic value of autoantibodies in lung cancer: a systematic review and meta-analysis[J]. Cell Physiol Biochem, 2018, 51(6): 2631-2646. doi: 10.1159/000495935 pmid: 30562746
[17]	NING Y C, HUI N, QING B, et al. ZCCHC10 suppresses lung cancer progression and cisplatin resistance by attenuating MDM2-mediated p53 ubiquitination and degradation[J]. Cell Death Dis, 2019, 10(6): 414. doi: 10.1038/s41419-019-1635-9 pmid: 31138778
[18]	SHAO L P, ZUO X L, YANG Y, et al. The inherited variations of a p53-responsive enhancer in 13q12.12 confer lung cancer risk by attenuating TNFRSF19 expression[J]. Genome Biol, 2019, 20(1): 103. doi: 10.1186/s13059-019-1696-1 pmid: 31126313
[19]	REN S X, ZHANG S C, JIANG T, et al. Early detection of lung cancer by using an autoantibody panel in Chinese population[J]. Oncoimmunology, 2018, 7(2): e1384108. doi: 10.1080/2162402X.2017.1384108
[20]	WEN W, LIU G, JIN K, et al. TGF-β1 induces PGP9.5 expression in CAFs to promote the growth of colorectal cancer cells[J]. Oncol Rep, 2017, 37(1): 115-122. doi: 10.3892/or.2016.5238 pmid: 27840994
[21]	LIN S C, CHOU Y T, JIANG S S, et al. Epigenetic switch between SOX2 and SOX9 regulates cancer cell plasticity[J]. Cancer Res, 2016, 76(23): 7036-7048. doi: 10.1158/0008-5472.CAN-15-3178
[22]	KAMEL L M, ATEF D M, MACKAWY A M H, et al. Circulating long non-coding RNA GAS5 and SOX2OT as potential biomarkers for diagnosis and prognosis of non-small cell lung cancer[J]. Biotechnol Appl Biochem, 2019, 66(4): 634-642. doi: 10.1002/bab.1764
[23]	TANG Z M, LING Z G, WANG C M, et al. Serum tumor-associated autoantibodies as diagnostic biomarkers for lung cancer: a systematic review and meta-analysis[J]. PLoS One, 2017, 12(7): e0182117. doi: 10.1371/journal.pone.0182117
[24]	MU Y Y, XIE F Y, WANG F B, et al. Performance evaluation of an enzyme-linked immunosorbent assay for seven autoantibodies in lung cancer[J]. Clin Lab, 2019, 65(4).
[25]	FANIPAKDEL A, SEILANIAN TOUSSI M, REZAZADEH F, et al. Overexpression of cancer-testis antigen melanoma-associated antigen A1 in lung cancer: a novel biomarker for prognosis, and a possible target for immunotherapy[J]. J Cell Physiol, 2019, 234(7): 12080-12086. doi: 10.1002/jcp.27884 pmid: 30569450

Item	p53	PGP9.5	SOX2	GAGE7	GBU4-5	MAGE A1
Tumor tissue	41 (41.0)	45 (45.0)	32 (32.0)	36 (36.0)	37 (37.0)	39 (39.0)
Adjacent tissue	3 (3.0)	4 (4.0)	2 (2.0)	1 (1.0)	1 (1.0)	1 (1.0)
Serum	35 (35.0)	46 (46.0)	30 (30.0)	34 (34.0)	35 (35.0)	40 (40.0)
χ² value	44.477	31.558	23.926	29.587	30.429	33.325
P value	<0.001	<0.001	<0.001	<0.001	<0.001	<0.001

Item	p53	PGP9.5	SOX2	GAGE7	GBU4-5	MAGE A1
Tumor tissue	0.52±0.13	0.48±0.14	0.39±0.11	0.35±0.12	0.29±0.08	0.42±0.19
Adjacent tissue	0.16±0.05	0.15±0.04	0.09±0.02	0.10±0.03	0.15±0.06	0.18±0.07
Serum	6.47±11.81	1.16±3.75	5.61±9.14	3.36±5.52	1.81±2.84	2.31±4.82
F value	5.038	8.137	5.121	4.389	5.009	5.001
P value	<0.001	<0.001	<0.001	<0.001	<0.001	<0.001