胃腺癌患者LHPP 表达及其对癌细胞生物学功能的影响

doi:10.3969/j.issn.1673-8640.2025.01.008

摘要/Abstract

摘要：

目的探讨胃腺癌患者磷酸赖氨酸磷酸组氨酸无机焦磷酸盐磷酸酶（LHPP ）表达的变化，及其对癌细胞生物学功能的影响。方法选取2020年10月—2021年6月江苏大学附属医院胃腺癌患者21例，收集其癌组织和癌旁组织。通过生物数据库分析LHPP 在胃癌中的表达及其与患者预后的关系。检测胃腺癌患者癌组织和正常胃组织，以及不同细胞株（人胃黏膜上皮细胞株GES-1和胃癌细胞株HGC-27、AGS、MKN-45、SGC-7901、MGC-803）中LHPP 蛋白和mRNA的表达情况。构建胃癌细胞干扰模型，并采用细胞生物学实验评估LHPP 对胃癌细胞增殖和迁移能力的影响。采用基因本体（GO）富集和京都基因与基因组数据库（KEGG）信号通路分析评估LHPP 相关基因的作用。结果癌症基因组图谱计划（TCGA）数据库、综合基因表达（GEO）数据库和人类蛋白质图谱（HPA）数据库的分析结果显示，胃癌组织LHPP mRNA和蛋白表达均低于癌旁组织（P<0.05）；Kaplan-Meier Plotter数据库的分析结果显示，LHPP 高表达的胃癌患者总生存期、首次进展生存期和再次进展生存期均长于LHPP 低表达的胃癌患者[风险比（HR）分别为0.66、0.57、0.64，95%可信区间（CI）分别为0.54~0.81、0.45~0.73、0.48~0.85]。各胃癌细胞LHPP 蛋白和mRNA相对表达量与GES-1细胞比较，差异均有统计学意义（P<0.05），选择AGS细胞进行后续实验。胃腺癌患者癌组织中LHPP 蛋白和mRNA相对表达量均低于癌旁组织（P<0.05）。LHPP 可抑制AGS细胞的增殖和迁移。LHPP 在胃癌中的主要功能为有丝分裂中的细胞周期调控、正向调控DNA代谢过程、蛋白折叠、调控核质转运等，主要涉及细胞周期、TPX2调控AURKA激活和表观遗传学调控基因表达等信号通路。结论胃腺癌患者LHPP 呈低表达，且与患者预后密切相关。LHPP 对胃腺癌细胞的增殖和迁移有抑制作用。

关键词: 磷酸赖氨酸磷酸组氨酸无机焦磷酸盐磷酸酶, 胃腺癌, 增殖, 迁移, 预后

Abstract:

Objective To investigate the role of phospholysine phosphohistidine inorganic pyrophosphate phosphatase （LHPP ） expression in patients with gastric adenocarcinoma and its effect on the biological function of carcinoma cells. Methods A total of 21 patients with gastric adenocarcinoma from the Affiliated Hospital of Jiangsu University from October 2020 to June 2021 were enrolled to collect gastric cancer tissues and adjacent tissues. The expression of LHPP in gastric cancer and its relationship with prognosis were analyzed by biological database. The expression of LHPP protein and mRNA in cancer tissues and adjacent tissues of patients with gastric cancer and different cell lines （human gastric mucosal epithelial cell line GES-1 and gastric cancer cell line HGC-27，AGS，MKN-45，SGC-7901，MGC-803） were determined. A gastric cancer cell interference model was constructed，and the effects of LHPP on the proliferation and migration of gastric cancer cells were evaluated by cell biology experiments. Gene Ontology （GO） enrichment and Kyoto Encyclopedia of Genes and Genomes（KEGG） signaling pathway analysis were used to evaluate the role of LHPP related genes. Results The analysis results of the Cancer Genome Atlas（TCGA），the Gene Expression Omnibus（GEO）and the Human Protein Atlas（HPA）databases showed that the LHPP mRNA and protein expressions in gastric cancer tissues were lower than those in adjacent tissues（P<0.05）. The analysis results of Kaplan-Meier Plotter database showed that the overall survival，first progression survival and reprogression survival of gastric cancer patients with high LHPP expression were longer than those with low LHPP expression [hazard ratios （HR） were 0.66，0.57 and 0.64，95% confidence intervals （CI） were 0.54-0.81，0.45-0.73 and 0.48-0.85，respectively]. The relative expression levels of LHPP protein and mRNA in all gastric cancer cell lines were statistically significant compared with GES-1 cells （P<0.05），and AGS cells were selected for follow-up tests. The relative expression levels of LHPP protein and mRNA in gastric cancer tissues were lower than those in adjacent tissues （P<0.05）. LHPP can inhibit the proliferation and migration of AGS cells. The main functions of LHPP in gastric cancer were cell cycle regulation in mitosis，positive regulation of DNA metabolism，protein folding，karyoplasmic transport and so on，mainly involved in cell cycle，TPX2 regulated AURKA activation，epigenetic regulation of gene expression and other signaling pathways. Conclusions The expression of LHPP is low in patients with gastric adenocarcinoma，which is related to the prognosis of patients，and LHPP has an inhibitory effect on the proliferation and migration of gastric adenocarcinoma cells.

Key words: Phospholysine phosphohistidine inorganic pyrophosphate phosphatase, Gastric carcinoma, Proliferation, Migration, Prognosis

中图分类号:

Q555

向敏, 舒扬, 盛玉程, 武晓白, 郭若楠, 蒋一凡. 胃腺癌患者LHPP 表达及其对癌细胞生物学功能的影响[J]. 检验医学, 2025, 40(1): 41-48.

XIANG Min, SHU Yang, SHENG Yucheng, WU Xiaobai, GUO Ruonan, JIANG Yifan. Expression of LHPP in patients with gastric adenocarcinoma and its effect on biological function of carcinoma cells[J]. Laboratory Medicine, 2025, 40(1): 41-48.

图/表 9

表1 引物序列

基因名称	上游引物（5'~3'）	下游引物（5'~3'）
GAPDH	ACCCAGAAGACTGTGGTGATGG	TTCAGCTCAGGGATGACCTT
LHPP	GCGACCATACCTGCTCATCC	CGTCTGCAATTACCACACAGTTT

图1 TCGA数据库和GEO数据库中胃癌组织和癌旁组织LHPP mRNA相对表达量比较注：（a）TCGA数据库中胃癌组织和癌旁组织LHPP mRNA表达的配对分析统计图；（b）TCGA 数据库中胃癌组织和癌旁组织LHPP mRNA表达的非配对分析统计图；（c）GEO数据库的GSE 19826数据集中胃癌组织和癌旁组织LHPP mRNA表达比较。

图2 HPA数据库中胃癌组织和癌旁组织LHPP 蛋白的表达注：（a）癌旁组织样本1；（b）癌旁组织样本2；（c）胃癌组织样本1；（d）胃癌组织样本2。

图3 不同LHPP mRNA表达情况胃癌患者Kaplan-Meier生存曲线注：（a）总生存期；（b）首次进展生存期；（c）再次进展生存期；—LHPP mRNA高表达；—LHPP mRNA低表达。

图4 不同细胞和胃腺癌患者癌组织、癌旁组织中LHPP 的表达情况注：（a）GES-1细胞和不同胃癌细胞LHPP 蛋白表达情况；（b）GES-1细胞和不同胃癌细胞LHPP mRNA相对表达量比较；与GES-1细胞比较，*P<0.05；（c）胃腺癌患者癌组织和癌旁组织LHPP mRNA相对表达量比较；（d）胃腺癌患者癌组织和癌旁组织LHPP 蛋白表达情况；1为癌旁组织，2为癌组织。

图5 sh-LHPP -AGS细胞LHPP 蛋白和mRNA的表达注：（a）LHPP 蛋白表达；（b）LHPP mRNA相对表达量；与对照AGS细胞比较，*P<0.05。

图6 LHPP 对不同AGS细胞增殖能力的影响注：（a）CCK-8实验；对照AGS细胞， sh-LHPP 1-AGS细胞， sh-LHPP 2-AGS细胞；（b）平板克隆实验；（c）不同细胞的集落形成数比较；与对照AGS细胞比较，*P<0.05。

图7 LHPP 对不同AGS细胞迁移能力的影响注：（a）Transwell实验；（b）不同细胞的侵袭数比较；（c）划痕实验；（d）不同细胞的相对划痕愈合率比较；与对照AGS细胞比较，*P<0.05。

图8 LHPP 相关基因及其GO富集和KEGG信号通路分析注：（a）LHPP 相关基因；（b）与LHPP 呈负相关的基因；（c）与 LHPP 呈正相关的基因；（d） LHPP 相关基因的GO富集和KEGG信号通路分析。

参考文献 18

[1]	SMYTH E C, NILSSON M, GRABSCH H I, et al. Gastric cancer[J]. Lancet, 2020, 396(10251):635-648. DOI PMID
[2]	LI G Z, DOHERTY G M, WANG J. Surgical management of gastric cancer:a review[J]. JAMA Surg, 2022, 157(5):446-454.
[3]	ZHANG B, WHITEAKER J R, HOOFNAGLE A N, et al. Clinical potential of mass spectrometry-based proteogenomics[J]. Nat Rev Clin Oncol, 2019, 16(4):256-268. DOI PMID
[4]	YOKOI F, HIRAISHI H, IZUHARA K. Molecular cloning of a cDNA for the human phospholysine phosphohistidine inorganic pyrophosphate phosphatase[J]. J Biochem, 2003, 133(5):607-614. PMID
[5]	HINDUPUR S K, COLOMBI M, FUHS S R, et al. The protein histidine phosphatase LHPP is a tumour suppressor[J]. Nature, 2018, 555(7698):678-682.
[6]	LIN J X, LIAN N Z, GAO Y X, et al. M6A methylation mediates LHPP acetylation as a tumour aerobic glycolysis suppressor to improve the prognosis of gastric cancer[J]. Cell Death Dis, 2022, 13(5):463.
[7]	LI C, YANG J, WANG W, et al. LHPP exerts a tumor-inhibiting role in glioblastoma via the downregulation of Akt and Wnt/β-catenin signaling[J]. J Bioenerg Biomembr, 2021, 53(1):61-71.
[8]	HOU B, LI W, XIA P, et al. LHPP suppresses colorectal cancer cell migration and invasion in vitro and in vivo by inhibiting Smad3 phosphorylation in the TGF-β pathway[J]. Cell Death Discov, 2021, 7(1):273.
[9]	PHAN J H, MOFFITT R A, STOKES T H, et al. Convergence of biomarkers,bioinformatics and nanotechnology for individualized cancer treatment[J]. Trends Biotechnol, 2009, 27(6):350-358.
[10]	MACHLOWSKA J, BAJ J, SITARZ M, et al. Gastric cancer:epidemiology,risk factors,classification,genomic characteristics and treatment strategies[J]. Int J Mol Sci, 2020, 21(11):4012.
[11]	DEBERARDINIS R J, LUM J J, HATZIVASSILIOU G, et al. The biology of cancer:metabolic reprogramming fuels cell growth and proliferation[J]. Cell Metab, 2008, 7(1):11-20.
[12]	TREPAT X, CHEN Z, JACOBSON K. Cell migration[J]. Compr Physiol, 2012, 2(4):2369-2392. DOI PMID
[13]	MAZZONE M, BERGERS G. Regulation of blood and lymphatic vessels by immune cells in tumors and metastasis[J]. Annu Rev Physiol, 2019, 81:535-560. DOI PMID
[14]	VALASTYAN S, WEINBERG R A. Tumor metastasis:molecular insights and evolving paradigms[J]. Cell, 2011, 147(2):275-292.
[15]	WU F, CHEN Y, ZHU J. LHPP suppresses proliferation,migration,and invasion and promotes apoptosis in pancreatic cancer[J]. Biosci Rep, 2020, 40(3):BSR20194142.
[16]	LI Y, ZHANG X, ZHOU X, et al. LHPP suppresses bladder cancer cell proliferation and growth via inactivating AKT/p65 signaling pathway[J]. Biosci Rep, 2019, 39(7):BSR20182270.
[17]	SUN W, QIAN K, GUO K, et al. LHPP inhibits cell growth and migration and triggers autophagy in papillary thyroid cancer by regulating the AKT/AMPK/mTOR signaling pathway[J]. Acta Biochim Biophys Sin(Shanghai), 2020, 52(4):382-389.
[18]	WANG X, CHENG H, ZHAO J, et al. Long noncoding RNA DLGAP1-AS2 promotes tumorigenesis and metastasis by regulating the Trim21/ELOA/LHPP axis in colorectal cancer[J]. Mol Cancer, 2022, 21(1):210.