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目的 明确山柰酚（Kaempferol）对人类T细胞白血病病毒1型（Human T-cell leukemia virus type-1, HTLV-1）和成人T细胞白血病/淋巴瘤（Adult T-cell leukemia/lymphoma, ATLL）的作用，探究其抗HTLV-1侵染及抑制ATLL增殖的分子机制。方法 采用细胞共培养及2种药物处理方式，通过流式细胞术、双荧光素酶报告基因技术、实时定量逆转录聚合酶链反应（RT-qPCR）和Western blot等方法探究对HTLV-1病毒侵染的影响及作用机制；采用CCK-8、克隆形成实验和流式细胞术检测山柰酚对ATLL细胞增殖和凋亡的影响，并通过网络药理学分析及分子实验验证山柰酚影响ATLL细胞的分子机制。结果 HTLV-1阳性细胞株与JETWT35细胞共培养后立即加入低剂量山柰酚，药物能够明显下调后者红色荧光蛋白的表达（P<0.01），抑制HTLV-1侵染JETWT35细胞（P<0.01）。HTLV-1阳性细胞株和Jurkat细胞共培养体系中立即加入低剂量山柰酚，药物能够明显下调WT-Luc启动子活力（P<0.01），并显著抑制受体细胞中HTLV-1病毒基因Tax、HBZ等转录水平（P<0.01），而病毒侵染完成（12 h）后再加入山柰酚，则无上述明显影响。机制上，低浓度山柰酚能够剂量依赖性抑制阳性细胞株ATL-T、ATL-2细胞表面包膜蛋白亚基gp46的表达（P<0.01）并抑制病毒颗粒释放，从而抑制HTLV-1侵染。此外，较高剂量山柰酚能够剂量依赖性抑制ATLL细胞株的恶性增殖并诱导细胞凋亡（P<0.01），且相较于HTLV-1阴性细胞，山柰酚对阳性细胞株的作用更为明显（P<0.01）。机制上，网络药理学显示山柰酚通过靶向关键靶点SRC和EGFR治疗ATLL，且Western blot证实山柰酚能够剂量依赖性抑制SRC和EGFR的表达，从而抑制ATLL增殖。结论 本研究首次揭示山奈酚具有双重作用：既能抑制HTLV-1病毒传播，又可阻断ATLL肿瘤细胞的恶性进展，表明其作为抗HTLV-1候选药物的潜在价值，为后续体内实验及临床转化研究提供了重要理论依据。
Abstract：Objective This study aimed to clarify the effects of kaempferol on human T-cell leukemia virus type-1(HTLV-1) and adult T-cell leukemia/lymphoma(ATLL), and to elucidate its molecular mechanisms in inhibiting HTLV-1 infection and suppressing ATLL proliferation. Methods Cell co-culture systems with two drug treatment regimens were established, and flow cytometry, dual-luciferase reporter assays, quantitative reverse transcription PCR(RT-qPCR), and Western blot were employed to investigate the effects of kaempferol on HTLV-1 infection and the underlying mechanisms. CCK-8 assays, colony formation assays, and flow cytometry were used to assess the effects of kaempferol on ATLL cell proliferation and apoptosis. In addition, network pharmacology combined with molecular validation experiments was performed to explore the signaling pathways involved. Results In co-cultures of HTLV-1-positive cell lines with JETWT35 cells, immediate low-dose kaempferol treatment significantly downregulated red fluorescent protein(RFP) expression(P < 0.01) and inhibited HTLV-1 infection(P<0.01). Likewise, in HTLV-1-positive/Jurkat co-cultures, low-dose kaempferol markedly reduced WT-Luc promoter activity(P<0.01) and significantly suppressed transcription of HTLV-1 viral genes, including Tax and HBZ(P<0.01). In contrast, administration of kaempferol after viral infection had been established(12 h post-infection) showed no such significant effect. Mechanistically, low concentrations of kaempferol dose-dependently suppressed expression of the envelope glycoprotein subunit gp46(P<0.01) in ATL-T and ATL-2 cells, thereby inhibiting viral particle release and subsequent HTLV-1 infection. At higher concentrations, kaempferol dose-dependently inhibited malignant proliferation and induced apoptosis in ATLL cell lines(P<0.01), with more pronounced effects observed in HTLV-1-positive cells compared to HTLV-1-negative cells(P<0.01). Network pharmacology analysis further suggested that kaempferol exerts its anti-ATLL effects by targeting key molecules SRC and EGFR, a finding validated by Western blot, which confirmed dose-dependently downregulation of SRC and EGFR expression. Conclusion In conclusion, this study demonstrates for the first time that kaempferol exerts dual actions: it inhibits HTLV-1 transmission and blocks malignant progression of ATLL cells. These findings underscore the potential of kaempferol as a candidate anti-HTLV-1 therapeutic agent and provide an important theoretical foundation for future in vivo studies and clinical translation.
[1]Giam CZ. HTLV-1 replication and adult T cell leukemia development[J]. Recent Results Cancer Res,2021, 217:209-243. DOI:10. 1007/978-3-030-57362-1_10.
[2]O’Donnell JS, Jaberolansar N, Chappell KJ. Human Tlymphotropic virus type 1 and antiretroviral therapy:practical considerations for pre-exposure and postexposure prophylaxis, transmission prevention, and mitigation of severe disease[J]. Lancet Microbe, 2024,5(4):e400-e408. DOI:10. 1016/S2666-5247(23)00359-2.
[3]Zuo X, Zhou R, Yang S, et al. HTLV-1 persistent infection and ATLL oncogenesis[J]. J Med Virol,2023, 95(1):e28424. DOI:10. 1002/jmv. 28424.
[4]Bangham CRM. HTLV-1 persistence and the oncogenesis of adult T-cell leukemia/lymphoma[J].Blood, 2023, 141(19):2299-2306. DOI:10. 1182/blood. 2022019332.
[5]Hleihel R, Akkouche A, Skayneh H, et al. Adult Tcell leukemia:a comprehensive overview on current and promising treatment modalities[J]. Curr Oncol Rep,2021, 23(12):141. DOI:10. 1007/s11912-021-01138-3.
[6]Rakha A, Umar N, Rabail R, et al. Anti-inflammatory and anti-allergic potential of dietary flavonoids:a review[J]. Biomed Pharmacother, 2022, 156:113945. DOI:10. 1016/j. biopha. 2022. 113945.
[7]Nejabati HR, Roshangar L. Kaempferol:a potential agent in the prevention of colorectal cancer[J]. Physiol Rep, 2022, 10(20):e15488. DOI:10. 14814/phy2. 15488.
[8]Imran M, Rauf A, Shah ZA, et al. Chemo-preventive and therapeutic effect of the dietary flavonoid kaempferol:a comprehensive review[J]. Phytother Res, 2019, 33(2):263-275. DOI:10. 1002/ptr. 6227.
[9]Periferakis A, Periferakis AT, Troumpata L, et al.Kaempferol:a review of current evidence of its antiviral potential[J]. Int J Mol Sci, 2023, 24(22):16299.DOI:10. 3390/ijms242216299.
[10]Yang ZF, Bai LP, Huang WB, et al. Comparison of in vitro antiviral activity of tea polyphenols against influenza A and B viruses and structure-activity relationship analysis[J]. Fitoterapia, 2014, 93:47-53.DOI:10. 1016/j. fitote. 2013. 12. 011.
[11]Furuta R, Yasunaga JI, Miura M, et al. Human T-cell leukemia virus type 1 infects multiple lineage hematopoietic cells in vivo[J]. PLoS Pathog, 2017, 13(11):e1006722. DOI:10. 1371/journal. ppat. 1006722.
[12]Kamoi K, Watanabe T, Uchimaru K, et al. Updates on HTLV-1 uveitis[J]. Viruses, 2022, 14(4):794. DOI:10. 3390/v14040794.
[13]Panfil AR, Al-Saleem J, Howard CM, et al. Stability of the HTLV-1 antisense-derived protein, HBZ, is regulated by the E3 ubiquitin-protein ligase, UBR5[J].Front Microbiol, 2018, 9:80. DOI:10. 3389/fmicb. 2018. 00080.
[14]Hachiman M, Yoshimitsu M, Ezinne C, et al. In vitro effects of arsenic trioxide, interferon α and zidovudine in adult T cell leukemia/lymphoma cells[J]. Oncol Lett,2018, 16(1):1305-1311. DOI:10. 3892/ol. 2018. 8771.
[15]Jeong HJ, Ryu YB, Park SJ, et al. Neuraminidase inhibitory activities of flavonols isolated from Rhodiola rosea roots and their in vitro anti-influenza viral activities[J]. Bioorg Med Chem, 2009, 17(19):6816-6823.DOI:10. 1016/j. bmc. 2009. 08. 036.
[16]Behbahani M, Sayedipour S, Pourazar A, et al. In vitro anti-HIV-1 activities of kaempferol and kaempferol-7-Oglucoside isolated from Securigera securidaca[J]. Res Pharm Sci, 2014, 9(6):463-469.
[17]Dai W, Bi J, Li F, et al. Antiviral efficacy of flavonoids against enterovirus 71 infection in vitro and in newborn mice[J]. Viruses, 2019, 11(7):625. DOI:10. 3390/v11070625.
[18]Care C, Sornjai W, Jaratsittisin J, et al. Discordant activity of kaempferol towards dengue virus and Japanese encephalitis virus[J]. Molecules, 2020, 25(5):1246.DOI:10. 3390/molecules25051246.
[19]Matsuoka M, Jeang KT. Human T-cell leukaemia virus type 1(HTLV-1)infectivity and cellular transformation[J]. Nat Rev Cancer, 2007, 7(4):270-280. DOI:10. 1038/nrc2111.
[20]Wolf S, Vercruyssen M, Cook L. HTLV-1-related adult T-cell leukemia/lymphoma:insights in early detection and management[J]. Curr Opin Oncol, 2022,34(5):446-453. DOI:10. 1097/CCO. 0000000000000883.
[21]Thompson MR, Kaminski JJ, Kurt-Jones EA, et al.Pattern recognition receptors and the innate immune response to viral infection[J]. Viruses, 2011, 3(6):920-940. DOI:10. 3390/v3060920.
[22]Carty M, Guy C, Bowie AG. Detection of viral infections by innate immunity[J]. Biochem Pharmacol,2021, 183:114316. DOI:10. 1016/j.bcp. 2020. 114316.
[23]Barnard AL, Igakura T, Tanaka Y, et al. Engagement of specific T-cell surface molecules regulates cytoskeletal polarization in HTLV-1–infected lymphocytes[J].Blood, 2005, 106(3):988-995. DOI:10. 1182/blood-2004-07-2850.
[24]Wallin M, Ekström M, Garoff H. Isomerization of the intersubunit disulphide-bond in Env controls retrovirus fusion[J]. EMBO J, 2004, 23(1):54-65. DOI:10. 1038/sj. emboj. 7600012.
[25]Weng CJ, Yen GC. Flavonoids, a ubiquitous dietary phenolic subclass, exert extensive in vitro anti-invasive and in vivo anti-metastatic activities[J]. Cancer Metastasis Rev, 2012, 31(1-2):323-351. DOI:10. 1007/s10555-012-9347-y.
[26]Neuhouser ML. Dietary flavonoids and cancer risk:evidence from human population studies[J]. Nutr Cancer, 2004, 50(1):1-7. DOI:10. 1207/s15327914nc5001_1.
[27]Azevedo C, Correia-Branco A, Araújo JR, et al. The chemopreventive effect of the dietary compound kaempferol on the MCF-7 human breast cancer cell line is dependent on inhibition of glucose cellular uptake[J].Nutr Cancer, 2015, 67(3):504-513. DOI:10. 1080/01635581. 2015. 1002625.
[28]Diantini A, Subarnas A, Lestari K, et al. Kaempferol-3-O-rhamnoside isolated from the leaves of Schima wallichii Korth. inhibits MCF-7 breast cancer cell proliferation through activation of the caspase cascade pathway[J]. Oncol Lett, 2012, 3(5):1069-1072.DOI:10. 3892/ol. 2012. 596.
[29]Riahi-Chebbi I, Souid S, Othman H, et al. The Phenolic compound Kaempferol overcomes 5-fluorouracil resistance in human resistant LS174 colon cancer cells[J]. Sci Rep, 2019, 9(1):195. DOI:10. 1038/s41598-018-36808-z.
[30]Yoshida T, Konishi M, Horinaka M, et al. Kaempferol sensitizes colon cancer cells to TRAIL-induced apoptosis[J]. Biochem Biophys Res Commun, 2008, 375(1):129-133. DOI:10. 1016/j. bbrc. 2008. 07. 131.
[31]Shuh M, Morse BA, Heidecker G, et al. Association of SRC-related kinase Lyn with the interleukin-2receptor and its role in maintaining constitutive phosphorylation of JAK/STAT in human T-cell leukemia virus type 1-transformed T cells[J]. J Virol,2011, 85(9):4623-4627. DOI:10. 1128/JVI. 00839-10.
[32]Kern D, Regl G, Hofbauer SW, et al. Hedgehog/GLI and PI3K signaling in the initiation and maintenance of chronic lymphocytic leukemia[J]. Oncogene, 2015, 34(42):5341-5351. DOI:10. 1038/onc. 2014. 450.
[33]Wee P, Wang Z. Epidermal growth factor receptor cell proliferation signaling pathways[J]. Cancers(Basel),2017, 9(5):52. DOI:10. 3390/cancers9050052.
[34]Joo EH, Bae JH, Park J, et al. Deconvolution of adult T-cell leukemia/lymphoma with single-cell RNA-seq using frozen archived skin tissue reveals new subset of cancer-associated fibroblast[J]. Front Immunol, 2022,13:856363. DOI:10. 3389/fimmu. 2022. 856363.
基本信息:
DOI：10.13242/j.cnki.bingduxuebao.250144
中图分类号:R965
引用信息:
[1]卯玉,方金勇,马婧,等.山柰酚抗HTLV‑1病毒侵染及抑制成人T细胞白血病增殖的机制研究[J].病毒学报,2026,42(02):493-505.DOI:10.13242/j.cnki.bingduxuebao.250144.
基金信息:
浙江省自然科学基金（项目号：ZCLMS25C0101）,题目：HTLV-1病毒通过调控细胞自噬抑制天然免疫病促进病毒复制的机制研究;浙江省自然科学基金（探索公益）（项目号：LT-LT24H080009）,题目：骨髓微环境通过S100A9诱导FLT3-ITD阳性AML细胞对奎扎替尼耐药的机制研究; 金华市科技计划项目（项目号：2026-3-003）,题目：HTLV-1病毒通过miR-155介导的细胞自噬途径促进白血病发生的分子机制~~
2025-09-23
2025-09-23
2025-09-23