热休克蛋白家族A成员1A在肝细胞癌中表达增高并介导索拉非尼耐药

摘要/Abstract

摘要： 目的: 探索热休克蛋白家族A成员1A（HSPA1A）在肝细胞癌（HCC）患者中表达及是否参与索拉非尼耐药机制。
方法: 选取青海省第五人民医院2022年3月至2024年3月收治的152例HCC患者。通过基因表达综合数据库中3个微阵列数据筛选介导索拉非尼耐药的枢纽基因。通过癌症基因组图谱-HCC队列、免疫组织化学（IHC）、实时定量聚合酶链反应（RT-qPCR）、蛋白质印迹（WB）分析及细胞功能等实验进一步探索耐药基因。通过药物敏感性分析HSPA1A表达与索拉非尼半数抑制浓度（IC50）相关性。通过单样本基因组富集基因分析（ssGSEA）算法分析HSPA1A表达与自噬信号间的关系。
结果: HSPA1A在索拉非尼治疗和抵抗组织中表达均显著增高（P<0.05）。IHC、RT-qPCR和WB证实索拉非尼抵抗患者HSPA1A表达水平均高于应答患者（均P<0.05）。基因表达谱互动分析数据库及IHC显示HCC组织中HSPA1A蛋白表达显著增高。临床队列及卡普尔-迈耶绘图仪数据库证实HSPA1A高表达患者总生存率和无复发生存率更低（均P<0.05）。CCK-8、克隆形成、划痕和侵袭实验证实敲降HSPA1A可显著抑制Huh-7和MHCC97-L细胞活力、增殖、迁移和侵袭能力（均P<0.05）。CCK-8实验结果显示索拉非尼在敲降HSPA1A的Huh-7和MHCC97-L细胞中的IC50分别降低17.4%和19.3%（均P<0.05）,增加索拉非尼治疗敏感性。ssGSEA分析显示HSPA1A mRNA高表达抑制自噬活性（P<0.05）。
结论: HSPA1A在HCC中表达增加并可能参与索拉非尼耐药机制,其潜在的机制可能与自噬信号有关。需要进一步实验探索HSPA1A在HCC中的潜在机制,为克服索拉非尼耐药提供新的策略。

关键词: 热休克蛋白家族A成员1A, 肝细胞癌, 索拉非尼, 耐药性, 自噬

Abstract: Objective: To explore the expression of heat shock protein family A member 1A (HSPA1A) in hepatocellular carcinoma (HCC) patients and whether it is involved in sorafenib resistance mechanism.
Methods: The 152 patients with HCC admitted to our hospital from March 2022 to March 2024 were selected. Hub genes mediating sorafenib resistance were screened by three microarray data from the Gene Expression Omnibus database. Resistance genes were further explored by The Cancer Genomic Atlas-HCC cohort, immunohistochemistry (IHC), quantitative real-time polymerase chain reaction (RT-qPCR), western blot (WB) analyses, and cellular function. The correlation between HSPA1A expression and sorafenib 50% inhibitory concentration(IC50) was analysed by drug sensitivity. The relationship between HSPA1A expression and autophagy signalling was analysed by single-sample single sample gene set enrichment analysis (ssGSEA) algorithm.
Results: HSPA1A expression was significantly higher in both sorafenib-treated and resistance tissues (P<0.05). IHC, RT-qPCR and WB confirmed that HSPA1A expression levels were higher in sorafenib-resistant patients than in responding patients (all P<0.05). Gene Expression Profiling Interactive Analysis database and IHC showed significantly higher HSPA1A protein expression in HCC tissues. Clinical cohorts and the Kaplan-Meier plotter database confirmed lower overall survival rate and relapse-free survival rate in patients with high HSPA1A expression (all P<0.05). CCK-8, clone formation, scratch and invasion assays confirmed that knockdown of HSPA1A significantly inhibited Huh-7 and MHCC97-L cell viability, proliferation, migration and invasion ability (all P<0.05).CCK-8 assay showed that the IC50 of sorafenib in Huh-7 and MHCC97-L cells knocked down by HSPA1A decreased by 17.4% and 19.3% (all P<0.05), increasing the therapeutic sensitivity of sorafenib. ssGSEA analysis showed that high HSPA1A mRNA expression inhibited autophagy activity (P<0.05).
Conclusion: HSPA1A expression is increased in HCC and may be involved in the mechanism of sorafenib resistance, and the underlying mechanism may be related to autophagy signalling. Further experiments are needed to explore the potential mechanism of HSPA1A in HCC and provide new strategies to overcome sorafenib resistance.

Key words: HSPA1A, Hepatocellular carcinoma, Sorafenib, Drug resistance, Autophagy

杨丽, 鲁雪. 热休克蛋白家族A成员1A在肝细胞癌中表达增高并介导索拉非尼耐药[J]. 肝癌电子杂志, 2025, 12(2): 14-21.

Yang Li, Lu Xue. Heat shock protein family A member 1A expression is increased in hepatocellular carcinoma and mediates sorafenib resistance[J]. Electronic Journal of Liver Tumor, 2025, 12(2): 14-21.

参考文献

[1] SON Y, SHIN N R, KIM S H, et al.Fibrinogen-like protein 1 modulates sorafenib resistance in human hepatocellular carcinoma cells[J]. Int J Mol Sci, 2021, 22(10): 5330.
[2] TANG W, CHEN Z, ZHANG W, et al.The mechanisms of sorafenib resistance in hepatocellular carcinoma: theoretical basis and therapeutic aspects[J]. Signal Transduct Target Ther, 2020, 5(1): 87.
[3] TAN X P, XIONG B H, ZHANG Y X, et al.FXYD5 promotes sorafenib resistance through the Akt/mTOR signaling pathway in hepatocellular carcinoma[J]. Eur J Pharmacol, 2022, 931: 175186.
[4] LU Y, CHAN Y T, TAN H Y, et al.Epigenetic regulation of ferroptosis via ETS1/miR-23a-3p/ACSL4 axis mediates sorafenib resistance in human hepatocellular carcinoma[J]. J Exp Clin Cancer Res, 2022, 41(1): 3.
[5] ZHU Y J, ZHENG B, WANG H Y, et al.New knowledge of the mechanisms of sorafenib resistance in liver cancer[J]. Acta Pharmacol Sin, 2017, 38(5): 614-622.
[6] MERY B, VALLARD A, ROWINSKI E, et al.High-throughput sequencing in clinical oncology: from past to present[J]. Swiss Med Wkly, 2019, 149: w20057.
[7] WU C, QI X, QIU Z, et al.Low expression of KIF20A suppresses cell proliferation, promotes chemosensitivity and is associated with better prognosis in HCC[J]. Aging (Albany NY), 2021, 13(18): 22148-22163.
[8] CHENG Z, WEI-QI J, JIN D.New insights on sorafenib resistance in liver cancer with correlation of individualized therapy[J]. Biochim Biophys Acta Rev Cancer, 2020, 1874(1): 188382.
[9] OGBODO E, MICHELANGELI F, WILLIAMS J H H. Exogenous heat shock proteins HSPA1A and HSPB1 regulate TNF-α, IL-1β and IL-10 secretion from monocytic cells[J]. FEBS Open Bio, 2023, 13(10): 1922-1940.
[10] RINGELHAN M, PFISTER D, O'CONNOR T, et al. The immunology of hepatocellular carcinoma[J]. Nat Immunol, 2018, 19(3): 222-232.
[11] WANG B, LAN T, XIAO H, et al.The expression profiles and prognostic values of HSP70s in hepatocellular carcinoma[J]. Cancer Cell Int, 2021, 21(1): 286.
[12] SUN T, LIU H, MING L.Multiple roles of autophagy in the sorafenib resistance of hepatocellular carcinoma[J]. Cell Physiol Biochem, 2017, 44(2): 716-727.
[13] RUSSELL R C, GUAN K L.The multifaceted role of autophagy in cancer[J]. EMBO J, 2022, 41(13): e110031.
[14] DEBNATH J, GAMMOH N, RYAN K M.Autophagy and autophagy-related pathways in cancer[J]. Nat Rev Mol Cell Biol, 2023, 24(8): 560-575.
[15] SHI Y H, DING Z B, ZHOU J, et al.Targeting autophagy enhances sorafenib lethality for hepatocellular carcinoma via ER stress-related apoptosis[J]. Autophagy, 2011, 7(10): 1159-1172.
[16] ZHAI B, HU F, JIANG X, et al.Inhibition of Akt reverses the acquired resistance to sorafenib by switching protective autophagy to autophagic cell death in hepatocellular carcinoma[J]. Mol Cancer Ther, 2014, 13(6): 1589-1598.
[17] TAI W T, SHIAU C W, CHEN H L, et al.Mcl-1-dependent activation of Beclin 1 mediates autophagic cell death induced by sorafenib and SC-59 in hepatocellular carcinoma cells[J]. Cell Death Dis, 2013, 4(2): e485.
[18] HE C, DONG X, ZHAI B, et al.MiR-21 mediates sorafenib resistance of hepatocellular carcinoma cells by inhibiting autophagy via the PTEN/Akt pathway[J]. Oncotarget, 2015, 6(30): 28867-28881.