自噬相关蛋白Beclin-1及微管相关蛋白1轻链3-Ⅱ在肺癌转移性胸腔积液中的表达研究

doi:10.12114/j.issn.1007-9572.2023.0812

中国全科医学 ›› 2026, Vol. 29 ›› Issue (05): 623-630.DOI: 10.12114/j.issn.1007-9572.2023.0812

自噬相关蛋白Beclin-1及微管相关蛋白1轻链3-Ⅱ在肺癌转移性胸腔积液中的表达研究

姚文静¹, 王翠峰²^,^*(), 高金亮³, 任美英², 景学芬², 付玉华²

1．014040　内蒙古自治区包头市，内蒙古科技大学包头医学院
2．014010　内蒙古自治区包头市，内蒙古科技大学包头医学院第一附属医院检验科
3．017000　内蒙古自治区鄂尔多斯市，鄂尔多斯中心医院分子医学实验室

收稿日期:2023-10-16 修回日期:2025-05-01 出版日期:2026-02-15 发布日期:2026-01-05
通讯作者: 王翠峰
作者贡献：
姚文静、高金亮负责进行实验，研究过程的实施；王翠峰、任美英提出研究命题、设计研究方案；王翠峰对文章整体负责；景学芬负责最终版本修订；付玉华负责统计学分析、绘制图表等。
基金资助:
内蒙古自然科学基金(2021MS08004)

Autophagy Related Proteins Beclin-1 and LC3-ⅡExpression in Lung Cancer Metaststic Pleural Effusion

YAO Wenjing¹, WANG Cuifeng²^,^*(), GAO Jinliang³, REN Meiying², JING Xuefen², FU Yuhua²

1. Baotou Medical College, Inner Mongolia University of Science and Technology, Baotou 014040, China
2. Department of Clinical Laboratory, the First Affiliated Hospital of Baotou Medical College, Inner Mongolia University of Science and Technology, Baotou 014010, China
3. Laboratory of Molecular Medicine, Ordos Central Hospital, Ordos 017000, China

Received:2023-10-16 Revised:2025-05-01 Published:2026-02-15 Online:2026-01-05
Contact: WANG Cuifeng

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摘要/Abstract

摘要： 背景恶性胸腔积液是肿瘤晚期以及转移的重要标志之一，现有诊断胸腔积液性质的相关方法存在缺陷与不足，无法对胸腔积液的性质进行及时有效的判断。多数恶性胸腔积液是由转移性肺癌引起，自噬相关蛋白1（Beclin-1）及用于监测细胞自噬水平的微管相关蛋白1轻链3（LC3）-Ⅱ在肺癌疾病进程中有重要作用，但在转移性胸腔积液中应用的相关报道较少，仍需进一步研究。目的分析自噬相关蛋白Beclin-1及LC3-Ⅱ在肺癌转移性胸腔积液中的表达情况，探讨其在临床疾病早期诊断中的价值。方法 2022年5月—2023年7月利用GEO数据库、GEPIA2及GeneMANIA数据库分析获得Beclin-1及LC3的生物信息；收集包头医学院第一附属医院检验科细胞室自2018—2023年未经任何临床治疗患者的胸腔积液，通过液基薄层细胞涂片并结合其临床资料对收集的胸腔积液进行分组，将其分为恶性测定组（95例）及良性对照组（190例），通过RT-PCR方法及Western Blotting方法分别检测Beclin-1、LC3-Ⅱ基因及蛋白水平表达，并利用免疫荧光试验确认LC3-Ⅱ蛋白的表达数量。结果生物信息学分析结果显示，Beclin-1与LC3-Ⅱ均存在于肺癌差异表达基因数据集中，在肺癌组织与正常组织中表达具有差异性；RT-PCR结果显示Beclin-1与LC3-Ⅱ的基因表达在良性对照组中均高于恶性测定组（P<0.05）；Western Blotting结果中Beclin-1与LC3-Ⅱ的蛋白表达在良性对照组较恶性测定组高（P<0.05）；此外在良性对照组中异硫氰酸荧光素标记的代表自噬体数量的LC3-Ⅱ荧光斑点较恶性测定组高。结论通过分析自噬相关蛋白Beclin-1及LC3-Ⅱ在良性胸腔积液及肺癌转移性恶性胸腔积液中的表达差异，可用于恶性胸腔积液的早期诊断，为良恶性胸腔积液的鉴别诊断与针对性治疗提供新的视角。

关键词: 肺肿瘤, 胸腔积液, 自噬, Beclin-1, LC3-Ⅱ

Abstract:

Background

Malignant pleural effusion, a critical clinical indicator of advanced-stage malignancy and metastatic progression. The existing methods for the diagnosis of the nature of pleural effusion have defects and shortcomings, and it is impossible to timely and effectively determine the nature of pleural effusion. Metastatic lung cancer constitutes the primary etiology of malignant pleural effusion. While the roles of autophagy-related proteins Beclin-1 and LC3-Ⅱ in lung cancer pathogenesis have been well established through multiple studies, their expression patterns and potential as diagnostic biomarkers in MPE urgently require further investigation.

Objective

To analyze the expression of autophagy-related proteins Beclin-1 and LC3-Ⅱ in lung cancer-associated malignant pleural effusion, and to evaluate their potential value in early clinical diagnosis.

Methods

From May 2022 to July 2023, we performed bioinformatics analysis of Beclin-1 and LC3 using GEO, GEPIA2, and GeneMANIA databases. Pleural effusion samples were collected from patients who had not received any clinical treatment at the Clinical Laboratory Cell Room of the First Affiliated Hospital of Baotou Medical College, Inner Mongolia University of Science and Technology, from 2018 to 2023. The collected pleural effusion samples were grouped through liquid-based thin-layer cytology smears combined with clinical data, dividing them into a malignant group (95 cases) and a benign control group (190 cases). The expression of Beclin-1 and LC3-Ⅱat the gene and protein levels was detected by RT-PCR and Western Blotting methods, respectively, and the expression level of LC3-Ⅱ protein was confirmed using immunofluorescence assay.

Results

Bioinformatics analysis confirmed the inclusion of Beclin-1 and LC3 in the differentially expressed gene profile of lung cancer. RT-PCR revealed significantly higher mRNA expression levels of both Beclin-1 and LC3-Ⅱ in benign controls versus malignant cases (P<0.05). Western Blotting analysis showed elevated Beclin-1 and LC3-Ⅱ protein abundance in benign specimens compared to malignant group (P<0.05). Immunofluorescence microscopy identified increased FITC-labeled LC3-Ⅱ puncta in benign controls.

Conclusion

By analyzing the expression differences of autophagy-related proteins Beclin-1 and LC3-Ⅱ in benign pleural effusion and metastatic malignant pleural effusion of lung cancer, may facilitate early diagnosis of MPE and provide novel perspectives for differential diagnosis and targeted therapeutic strategies.

Key words: Lung neoplasms, Pleural effusion, Autophgy, Beclin-1, LC3-Ⅱ

中图分类号:

R 734.2

姚文静,王翠峰,高金亮,等. 自噬相关蛋白Beclin-1及微管相关蛋白1轻链3-Ⅱ在肺癌转移性胸腔积液中的表达研究[J]. 中国全科医学, 2026, 29(05): 623-630. DOI: 10.12114/j.issn.1007-9572.2023.0812.
YAO Wenjing,WANG Cuifeng,GAO Jinliang, et al. Autophagy Related Proteins Beclin-1 and LC3-ⅡExpression in Lung Cancer Metaststic Pleural Effusion[J]. Chinese General Practice, 2026, 29(05): 623-630. DOI: 10.12114/j.issn.1007-9572.2023.0812.

图/表 9

图1 Beclin-1及LC3的相互作用蛋白网络注：A为自噬相关蛋白1（Beclin-1）的相互作用蛋白网络；B为微管相关蛋白1轻链3（LC3）的相互作用蛋白网络。CASP10=半胱天冬酶10，BCL2L1=B细胞淋巴瘤2样蛋白1，AMBRA1=Beclin1调节因子1，UVRAG=紫外线抗性相关基因，PIK3R4=磷酸肌醇3-激酶调节亚基4，TSC1=结节性硬化症复合体1，NRBF2=核受体结合因子2，SQSTM1=自噬接头蛋白，RETREG1=网状自噬调节因子1，Network=网络，Physical Interactions=物理相互作用，Co-expression=共表达，Predicted=预测关联，Co-localization=共定位，Genetic Interactions=遗传互作，Pathway=通路，Shared protein domains=共享蛋白结构域，phosphatidylinositol 3-kinase complex=磷脂酰肌醇3-激酶复合体，macronutophagy=巨自噬，cellular response to nutrient levels=细胞对营养水平的响应，cellular response to starvation=细胞对饥饿的响应，cellular response to extracellular stimulus=细胞对胞外刺激的响应，autophagosome organization=自噬体组装，regulation of cell division=细胞分裂调控，autophagosome=自噬体。

Figure 1 Diagram of the interacting protein network of Beclin-1 and LC3

表1 GSE19188数据库Beclin-1、LC3相关参数

Table 1 Beclin-1 and LC3 related parameters in the GSE19188 database

基因名称	校正后P值	P值
Beclin-1	1.49×10^-2	8.45×10^-3
LC3	6.44×10^-3	3.32×10^-3

图2 差异基因的火山图注：红点代表差异基因中表达上调的基因，蓝点代表差异基因中表达下调的基因，黑色为差异基因中无明显变化的基因。tumor=肿瘤，health=健康，P value=P值，fold change=倍数变化。

Figure 2 Volcano map of differential genes

图3 良恶性胸腔积液显微镜下细胞形态（×100）注：A表示良性胸腔积液，B表示恶性胸腔积液。

Figure 3 Microscopic cell morphology of benign and malignant pleural effusion

表2 PCR引物序列表

Table 2 PCR primer sequence table

基因名称	引物序列
Beclin-1	F	5'-CTGGACACTCAGCTCAACGTCA-3'
Beclin-1	R	5'-CTCTAGTGCCAGCTCCTTTAGC-3'
LC3	F	5'-GCTACAAGGGTGAGAAGCAGCT-3'
LC3	R	5'-CTGGTTCACCAGCAGGAAGAAG-3'
β-actin	F	5'-TCCCTGGAGAAGAGCTACGA-3'
β-actin	R	5'-AGCACTGTGTTGGCGTACAG-3'

图4 GEPIA2分析TCGA数据库中Beclin-1 、LC3 mRNA表达注：A表示Beclin-1的mRNA表达结果分析图，B表示LC3的mRNA表达结果分析图；LUAD=肺腺癌，LUSC=肺鳞癌。

Figure 4 GEPIA2 analyzed Beclin-1，LC3 mRNA expression in the TCGA database

图5 良恶性胸腔积液中Beclin-1/LC3-ⅡmRNA相对表达量注：A为Beclin-1 mRNA的相对表达量；B为LC3-Ⅱ mRNA的相对表达量；a表示P<0.05，b表示P<0.01。

Figure 5 The expression levels of Beclin-1/LC3-Ⅱ mRNA in benign/malignant pleural effusion

图6 良恶性胸腔积液中Beclin-1、LC3-Ⅱ蛋白表达量注：A表示Western Blotting目的基因蛋白Beclin-1表达条带；B表示Western Blotting目的基因蛋白LC3-Ⅱ表达条带；C表示Beclin-1蛋白的相对表达量；D表示LC3-Ⅱ蛋白的相对表达量；a表示P<0.01。

Figure 6 The expression levels of Beclin-1/LC3-Ⅱ protein in benign/malignant pleural effusion

图7 荧光显微镜下良恶性胸腔积液LC3-Ⅱ表达量注：A表示良性胸腔积液（×10），B表示良性胸腔积液（×100），C表示恶性胸腔积液（×10），D表示恶性胸腔积液（×100）。

Figure 7 Expression of LC3-Ⅱ of benign /malignant pleural effusion under fluorescence microscopy

参考文献 35

[1]	BRAY F, FERLAY J, SOERJOMATARAM I, et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2018, 68(6): 394-424. DOI: 10.3322/caac.21492.
[2]	CARTER J, MILLER J A, FELLER-KOPMAN D, et al. Molecular profiling of malignant pleural effusion in metastatic non-small-cell lung carcinoma. the effect of preanalytical factors[J]. Ann Am Thorac Soc, 2017, 14(7): 1169-1176.
[3]	XIA H W, ZHANG Z Q, YUAN J, et al. Human RECQL5 promotes metastasis and resistance to cisplatin in non-small cell lung cancer[J]. Life Sci, 2021, 265: 118768. DOI: 10.1016/j.lfs.2020.118768.
[4]	HOWLADER N, FORJAZ G, MOORADIAN M J, et al. The effect of advances in lung-cancer treatment on population mortality[J]. N Engl J Med, 2020, 383(7): 640-649.
[5]	CHEN R Q, MANOCHAKIAN R, JAMES L, et al. Emerging therapeutic agents for advanced non-small cell lung cancer[J]. J Hematol Oncol, 2020, 13(1): 58.
[6]	PSALLIDAS I, KALOMENIDIS I, PORCEL J M, et al. Malignant pleural effusion: from bench to bedside[J]. Eur Respir Rev, 2016, 25(140): 189-198. DOI: 10.1183/16000617.0019-2016.
[7]	GAYEN S. Malignant pleural effusion: presentation, diagnosis, and management[J]. Am J Med, 2022, 135(10): 1188-1192.
[8]	BIBBY A C, DORN P, PSALLIDAS I, et al. ERS/EACTS statement on the management of malignant pleural effusions[J]. Eur Respir J, 2018, 52(1): 1800349. DOI: 10.1183/13993003.00349-2018.
[9]	FENG Y C, HE D, YAO Z Y, et al. The machinery of macroautophagy[J]. Cell Res, 2014, 24(1): 24-41.
[10]	HANAHAN D, WEINBERG R A. Hallmarks of cancer: the next generation[J]. Cell, 2011, 144(5): 646-674.
[11]	ALZAHRANI A S. PI3K/Akt/mTOR inhibitors in cancer: At the bench and bedside[J]. Semin Cancer Biol, 2019, 59: 125-132. DOI: 10.1016/j.semcancer.2019.07.009.
[12]	LI X H, HE S K, MA B Y. Autophagy and autophagy-related proteins in cancer[J]. Mol Cancer, 2020, 19(1): 12.
[13]	JACQUET M, GUITTAUT M, FRAICHARD A, et al. The functions of Atg8-family proteins in autophagy and cancer: linked or unrelated [J]. Autophagy, 2021, 17(3): 599-611.
[14]	POON I K, CHAN R C K, CHOI J S H, et al. A comparative study of diagnostic accuracy in 3026 pleural biopsies and matched pleural effusion cytology with clinical correlation[J]. Cancer Med, 2023, 12(2): 1471-1481. DOI: 10.1002/cam4.5038.
[15]	ARNOLD D T, DE FONSEKA D, PERRY S, et al. Investigating unilateral pleural effusions: the role of cytology[J]. Eur Respir J, 2018, 52(5): 1801254. DOI: 10.1183/13993003.01254-2018.
[16]	WANG S F, TIAN S, LI Y, et al. Development and validation of a novel scoring system developed from a nomogram to identify malignant pleural effusion[J]. EBioMedicine, 2020, 58: 102924. DOI: 10.1016/j.ebiom.2020.102924.
[17]	WOO C G, SON S M, HAN H S, et al. Diagnostic benefits of the combined use of liquid-based cytology, cell block, and carcinoembryonic antigen immunocytochemistry in malignant pleural effusion[J]. J Thorac Dis, 2018, 10(8): 4931-4939.
[18]	HAN Y H, MUN J G, JEON H D, et al. Betulin inhibits lung metastasis by inducing cell cycle arrest, autophagy, and apoptosis of metastatic colorectal cancer cells[J]. Nutrients, 2019, 12(1): 66. DOI: 10.3390/nu12010066.
[19]	ZHANG S Q, QIAN Y, YE L H. Delineating the twin role of autophagy in lung cancer[J]. Biol Futura, 2023, 74(1): 119-135. DOI: 10.1007/s42977-023-00165-4.
[20]	YU J E, KIM Y, HONG D E, et al. Bee venom triggers autophagy-induced apoptosis in human lung cancer cells via the mTOR signaling pathway[J]. J Oncol, 2022, 2022: 8916464.
[21]	RAGUSA M A, NASELLI F, CRUCIATA I, et al. Indicaxanthin induces autophagy in intestinal epithelial cancer cells by epigenetic mechanisms involving DNA methylation[J]. Nutrients, 2023, 15(15): 3495. DOI: 10.3390/nu15153495.
[22]	LI X, YANG K B, CHEN W, et al. CUL3(cullin 3)-mediated ubiquitination and degradation of BECN1(beclin 1)inhibit autophagy and promote tumor progression[J]. Autophagy, 2021, 17(12): 4323-4340. DOI: 10.1080/15548627.2021.1912270.
[23]	NICCO C, THOMAS M, GUILLERMET J, et al. Mechanistic target of rapamycin (mTOR) regulates self-sustained quiescence, tumor indolence, and late clinical metastasis in a Beclin-1-dependent manner[J]. Cell Cycle, 2023, 22(5): 542-564.
[24]	DI LEO L, BODEMEYER V, BOSISIO F M, et al. Loss of Ambra1 promotes melanoma growth and invasion[J]. Nat Commun, 2021, 12(1): 2550.
[25]	WAN Y H, QIAN Y H, WANG Y Y, et al. Prognostic value of Beclin 1, EGFR and ALK in non-squamous non-small cell lung cancer[J]. Discov Oncol, 2022, 13(1): 127.
[26]	刘俊英, 杨会钗, 李诗, 等. 肺腺癌中CALHM2、PD-L1的表达及其预后分析[J]. 临床与实验病理学杂志, 2023, 39(8): 957-962. DOI: 10.13315/j.cnki.cjcep.2023.08.013.
[27]	LIANG L J, HUI K Y, HU C X, et al. Autophagy inhibition potentiates the anti-angiogenic property of multikinase inhibitor anlotinib through JAK2/STAT3/VEGFA signaling in non-small cell lung cancer cells[J]. J Exp Clin Cancer Res, 2019, 38(1): 71. DOI: 10.1186/s13046-019-1093-3.
[28]	ICHIMURA Y, KOMINAMI E, TANAKA K, et al. Selective turnover of p62/A170/SQSTM1 by autophagy[J]. Autophagy, 2008, 4(8): 1063-1066. DOI: 10.4161/auto.6826.
[29]	SUN X D, LI J, LI Y Z, et al. Apatinib, a novel tyrosine kinase inhibitor, promotes ROS-dependent apoptosis and autophagy via the Nrf2/HO-1 pathway in ovarian cancer cells[J]. Oxid Med Cell Longev, 2020, 2020: 3145182. DOI: 10.1155/2020/3145182.
[30]	XING Y H, WEI X Q, LIU Y C, et al. Autophagy inhibition mediated by MCOLN1/TRPML1 suppresses cancer metastasis via regulating a ROS-driven TP53/p53 pathway[J]. Autophagy, 2022, 18(8): 1932-1954.
[31]	JIANG X W, LU W Q, SHEN X Y, et al. Repurposing sertraline sensitizes non-small cell lung cancer cells to erlotinib by inducing autophagy[J]. JCI Insight, 2018, 3(11): e98921.
[32]	LV Y S, ZHANG W J, ZHAO J Y, et al. SRSF1 inhibits autophagy through regulating Bcl-x splicing and interacting with PIK3C3 in lung cancer[J]. Signal Transduct Target Ther, 2021, 6(1): 108. DOI: 10.1038/s41392-021-00495-6.
[33]	王婧喆, 王翠峰. Beclin-1、LC3和mTOR在肺癌中的表达及意义[J]. 包头医学院学报, 2022, 38(1): 45-49. DOI: 10.16833/j.cnki.jbmc.2022.01.011.
[34]	TORII S, HONDA S, MUROHASHI M, et al. Autophagy involvement in oncogenesis[J]. Cancer Sci, 2020, 111(11): 3993-3999. DOI: 10.1111/cas.14646.
[35]	WU H, LU X X, WANG J R, et al. TRAF6 inhibits colorectal cancer metastasis through regulating selective autophagic CTNNB1/β-catenin degradation and is targeted for GSK3B/GSK3β-mediated phosphorylation and degradation[J]. Autophagy, 2019, 15(9): 1506-1522. DOI: 10.1080/15548627.2019.1586250.

自噬相关蛋白Beclin-1及微管相关蛋白1轻链3-Ⅱ在肺癌转移性胸腔积液中的表达研究

Autophagy Related Proteins Beclin-1 and LC3-ⅡExpression in Lung Cancer Metaststic Pleural Effusion

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