腺苷酸活化蛋白激酶在哈萨克族食管鳞癌组织中的表达及意义

doi:10.3969/j.issn.1004-616x.2023.04.007

摘要/Abstract

摘要： 目的：研究腺苷酸活化蛋白激酶(AMPK)在哈萨克族食管鳞癌患者癌组织中的表达水平，及其与患者临床病理指标和预后的关系，探讨AMPK在哈萨克族食管鳞癌脂质代谢重编程中的作用。方法：选取65例哈萨克族食管鳞癌患者。采用qPCR方法检测食管鳞癌及癌旁正常组织中AMPK mRNA的表达水平。免疫组织化学法检测食管鳞癌组织及癌旁正常组织中AMPK蛋白表达水平。Kaplan-Meier法绘制生存曲线分析食管鳞癌组织中AMPK表达与患者预后的相关性。Cox比例风险回归分析影响食管鳞癌患者不良预后发生的危险因素。在食管癌细胞株KYSE150中稳定敲低AMPK，通过超高效液相色谱-质谱法进行脂质定性定量分析。结果：食管鳞癌组织中AMPK的mRNA及蛋白表达水平高于癌旁组织(P<0.05)。AMPK表达水平与癌组织分化程度、淋巴结转移情况、TNM分期相关(P<0.05)，AMPK的阳性表达是哈萨克族食管鳞癌患者不良预后的危险因素。通过对食管癌细胞脂质含量的定量分析，发现不同脂质等级中脂质含量及变化趋势不同。共鉴定出10种脂质亚类，由脂肪酸类(7.88%)、甘油脂类(60.77%)、甘油鞘脂类(21.35%)、鞘脂类(9.62%)和固醇脂类(0.38%)组成。AMPK诱导食管癌细胞发生明显的脂质代谢变化，共发现76个差异脂质代谢物，其中37个上调，29个下调，其中甘油三酯(TAG)上调和鞘磷脂(SM)、甘油二酯(DAG)和磷脂酰乙醇胺(PE)下调，推测这些分布的变化有助于阐明食管癌中脂质积累的原因和可能的致癌分子机制。结论：AMPK在食管鳞癌组织中高表达，可能参与了食管鳞癌的发生与发展，对食管鳞癌的预后具有重要意义；AMPK可能通过介导脂质代谢重编程参与哈萨克族食管鳞癌的进展。

关键词: 食管鳞癌, 哈萨克族, 腺苷酸活化蛋白激酶, 生存预后, 代谢重编程

Abstract: OBJECTIVE： To investigate expression levels of the AMPK in Kazakh esophageal squamous cell carcinoma，its role in lipid metabolism reprogramming，and its relationship with clinicopathological features and prognosis. METHODS： Sixty-five Kazakh patients with esophageal squamous cell carcinomas were selected. AMPK mRNA expression levels in the cancer cells and the adjacent normal tissues were detected using qPCR. AMPK protein expressions were detected using immunohistochemistry. Kaplan-Meier method was used to draw the survival curve，and to analyze the correlation between AMPK expression and the prognosis of the carcinomas. Cox proportional hazards regression analysis was used to analyze the risk factors affecting the poor prognosis of the cancer patients. AMPK was stably knocked down in the esophageal cancer cell line，KYSE150，and lipid qualitative and quantitative analyses were performed using ultra performance liquid chromatography-mass spectrometry. RESULTS： The mRNA levels and protein positive rates of AMPK in the carcinoma tissues were significantly higher than those in adjacent tissues (P<0.05). The expression level of AMPK was correlated with the degree of differentiation，lymph node metastasis and TNM stage (P<0.05). The expression levels of AMPK were correlated with the degrees of differentiation，lymph node metastasis and TNM stages (P<0.05). The positive expression of AMPK was a reliable risk factor for poor prognosis of esophageal squamous cell carcinoma among Kazakhs. Through the absolute quantitative analysis of lipid content in esophageal cancer cells，it was found that the lipid content and its changing trend were different in different lipid grades. A total of 10 lipid subclasses were identified，consisting of fatty acids (7.88%)，glycerides (60.77%)，glycerides sphingolipids (21.35%)，sphingolipids (9.62%) and sterol lipids (0.38%). AMPK induced significant changes in lipid metabolism in esophageal cancer cells. A total of 76 differential lipid metabolites were found，among which 37 were up-regulated and 29 were down-regulated. The up-regulation of triglyceride (TAG) and the down-regulation of sphingolipid (SM)，diglyceride (DAG) and phosphatidyl ethanolamine (PE) were observed. Targeted lipid quantitative analysis results suggest that AMPK might be involved in lipid metabolic reprogramming in Kazakh patients with esophageal squamous cell carcinoma. CONCLUSION： The mRNA level and protein positive rate of AMPK in esophageal squamous cell carcinoma tissues were significantly higher than those in adjacent tissues (P<0.05). The expression level of AMPK was correlated with the degree of differentiation，lymph node metastasis and TNM stage (P<0.05). The positive expression of AMPK was a reliable risk factor for poor prognosis of Kazakh esophageal squamous cell carcinoma.

Key words: esophageal squamous cell carcinoma, Kazakh, AMPK, survival prognosis, metabolic reprogramming

中图分类号:

R735.1

刘瑞雪, 张力为. 腺苷酸活化蛋白激酶在哈萨克族食管鳞癌组织中的表达及意义[J]. 癌变·畸变·突变, 2023, 35(4): 285-291.

LIU Ruixue, ZHANG Liwei. Expression and significance of AMPK esophageal squamous cell carcinoma among Kazakhs[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2023, 35(4): 285-291.

参考文献

[1] MORGAN E, SOERJOMATARAM I, RUMGAY H, et al. The global landscape of esophageal squamous cell carcinoma and esophageal adenocarcinoma incidence and mortality in 2020 and projections to 2040: new estimates from GLOBOCAN 2020[J]. Gastroenterology, 2022, 163(3): 649-658.e2.
[2] UHLENHOPP D J, THEN E O, SUNKARA T, et al. Epidemiology of esophageal cancer: update in global trends, etiology and risk factors[J]. Clin J Gastroenterol, 2020, 13(6): 1010-1021.
[3] ZHENG S T, VUITTON L, SHEYHIDIN I, et al. Northwestern China: a place to learn more on oesophageal cancer. Part two: gene alterations and polymorphisms[J]. Eur J Gastroenterol Hepatol, 2011, 23(12): 1087-1099.
[4] ZHENG S T, HUO Q, TUERXUN A, et al. The expression and activation of ERK/MAPK pathway in human esophageal cancer cell line EC9706[J]. Mol Biol Rep, 2011, 38(2): 865-872.
[5] MENENDEZ J A, LUPU R. Fatty acid synthase and the lipogenic phenotype in cancer pathogenesis[J]. Nat Rev Cancer, 2007, 7(10): 763-777.
[6] SALMINEN A, KAARNIRANTA K, KAUPPINEN A. AMPK and HIF signaling pathways regulate both longevity and cancer growth: the good news and the bad news about survival mechanisms[J]. Biogerontology, 2016, 17(4): 655-680.
[7] GANAPATHY-KANNIAPPAN S, GESCHWIND J F. Tumor glycolysis as a target for cancer therapy: progress and prospects[J]. Mol Cancer, 2013, 12: 152.
[8] DASGUPTA B, CHHIPA R R. Evolving lessons on the complex role of AMPK in normal physiology and cancer[J]. Trends Pharmacol Sci, 2016, 37(3): 192-206.
[9] 郭幸, 刘宗文, 张艳, 等. 食管鳞癌组织中AMPKα1及磷酸化AMPKα的表达[J]. 郑州大学学报: 医学版, 2017, 52(3): 255-258.
[10] HERZIG S, SHAW R J. AMPK: guardian of metabolism and mitochondrial homeostasis[J]. Nat Rev Mol Cell Biol, 2018, 19(2): 121-135.
[11] WANG Q, LIU S D, ZHAI A H, et al. AMPK-mediated regulation of lipid metabolism by phosphorylation[J]. Biol Pharm Bull, 2018, 41(7): 985-993.
[12] GAO L X, XU Z G, HUANG Z, et al. CPI-613 rewires lipid metabolism to enhance pancreatic cancer apoptosis via the AMPK-ACC signaling[J]. J Exp Clin Cancer Res, 2020, 39(1): 73.
[13] ZHANG Z G, ZHANG H S, SUN H L, et al. KDM5B promotes breast cancer cell proliferation and migration via AMPK-mediated lipid metabolism reprogramming[J]. Exp Cell Res, 2019, 379(2): 182-190.
[14] LOPE V, GUERRERO-ZOTANO á, CASAS A, et al. Serum phospholipids fatty acids and breast cancer risk by pathological subtype[J]. Nutrients, 2020, 12(10): 3132.
[15] XU D Q, WANG Z, XIA Y, et al. The gluconeogenic enzyme PCK1 phosphorylates INSIG1/2 for lipogenesis[J]. Nature, 2020, 580(7804): 530-535.
[16] QU Y Y, ZHAO R, ZHANG H L, et al. Inactivation of the AMPK-GATA3-ECHS1 pathway induces fatty acid synthesis that promotes clear cell renal cell carcinoma growth[J]. Cancer Res, 2020, 80(2): 319-333.
[17] LIU J H, WU Q F, FU J K, et al. Obesity potentiates esophageal squamous cell carcinoma growth and invasion by AMPK-YAP pathway[J]. J Immunol Res, 2020, 2020: 6765474.