肺癌组织和外周血中p53、PGP9.5、SOX2、GAGE7、GBU4-5和MAGE A1蛋白水平检测及其临床价值探讨

doi:10.19401/j.cnki.1007-3639.2023.01.004

摘要/Abstract

摘要：

背景与目的：肺癌发病机制具有多样性，但一经确认，往往已错过最佳治疗时机，本文通过探讨肺癌组织和外周血中抑癌基因（p53）、蛋白基因产物（PGP9.5）、转录因子（SOX2）、肿瘤相关基因编码蛋白（GAGE7）、解旋酶（GBU4-5）和黑色素瘤抗原（MAGE A1）蛋白水平的相关性，同时分析其在肺癌诊疗中的应用价值。方法：回顾并分析2018年5月—2020年5月在复旦大学附属肿瘤医院确诊的100例肺癌患者的病历资料，临床TNM分期Ⅰ期25例，Ⅱ期45例，Ⅲa期30例；非小细胞肺癌80例，小细胞肺癌20例；取手术切除的肿瘤组织和癌旁组织，采用免疫组织化学染色法检测上述6种蛋白的水平，采用实时荧光定量聚合酶链反应（real-time fluorescence quantitative polymerase chain reaction，RTFQ-PCR）法检测其基因表达，采用酶联免疫吸附实验（enzyme-linked immunosorbent assay，ELISA）检测外周血中6种蛋白的抗体阳性情况。结果：肿瘤组织中p53、PGP9.5、SOX2、GAGE7、GBU4-5和MAGE A1蛋白的阳性率和基因表达水平均明显高于癌旁组织（P<0.05）；将其与肿瘤的临床病理学特征进行相关分析发现，肿瘤组织和外周血中的p53、PGP9.5、SOX2、GAGE7、GBU4-5和MAGE A1蛋白的阳性率和基因表达水平与肿瘤TNM分期和分化级别有关（P <0.05），而与患者性别、年龄、肿瘤直径、病理学类型无关（P>0.05）。肿瘤组织中6种蛋白的表达水平与外周血清表达具有较好的一致性（P>0.05）。结论：肺癌肿瘤组织和外周血中p53、PGP9.5、SOX2、GAGE7、GBU4-5和MAGE A1蛋白表达阳性与肿瘤TNM分期及分化级别密切相关。

关键词: 肺癌, 抑癌基因p53, 蛋白基因产物（PGP9.5）, 转录因子（SOX2）, 肿瘤相关基因编码蛋白（GAGE7）, 解旋酶（GBU4-5）, 黑色素瘤抗原（MAGE A1）

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)

中图分类号:

R734.2

邹淳缘, 许晓峰, 卢仁泉, 郭林. 肺癌组织和外周血中p53、PGP9.5、SOX2、GAGE7、GBU4-5和MAGE A1蛋白水平检测及其临床价值探讨[J]. 中国癌症杂志, 2023, 33(1): 36-44.

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.

图/表 6

表1

表2

表3

表4

肿瘤组织蛋白阳性表达和相对表达水平与肿瘤临床病理学特征的关系"

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)

表4

表5

血清中p53、PGP9.5、SOX2、GAGE7、GBU4-5、MAGE A1阳性率及表达水平与肿瘤临床病理学特征的关系"

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)

表5

图1

参考文献 25

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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