Anti-PF4 antibody inhibit proliferation of lung cancer cells without TP53 mutation by inducing apoptosis and cell cycle arrest

Abstract

Background: Elevated platelet count is correlated with poor survival of lung cancer. Platelet factor 4 (PF4), a platelet-specific protein, plays an important role in platelet function. Emerging evidence suggests that regulation of PF4 can modulate platelet function to impact the progression of lung cancer.

Methods: Anti-PF4 antibody was used to neutralise PF4 to regulate platelet function in the co-culture of platelets and lung cancer cells lines of H1299 and A549. The proliferation, apoptosis and cell cycle were examined by cell counting kit-8 (CCK-8) test and flow cytometry.

Results: Anti-PF4 antibody inhibited the proliferation of H1299 and A549 cells when stimulated with platelets, yet induced cell cycle arrest and cell apoptosis only in A549, not H1299. The proliferation, cell cycle distribution and apoptosis were not affected by anti-PF4 antibody in A549^TP53‒ cells.

Conclusion: Anti-PF4 antibody exerted anti-proliferative effects by inducing apoptosis and cell cycle arrest in lung cancer cells without TP53 mutation via p53 signalling pathway when stimulated with platelets.

Keywords

lung cancer / PF4 / platelet / platelet factor 4 / TP53

References

[1] Thai AA, Solomon BJ, Sequist LV, Gainor JF, Heist RS. Lung cancer. Lancet. 2021; 398(10299): 535-554.

[2] Siegel RL, Kratzer TB, Giaquinto AN, Sung H, Jemal A. Cancer statistics, 2025. CA Cancer J Clin. 2025; 75(1): 10-45.

[3] Bailey SE, Ukoumunne OC, Shephard EA, Hamilton W. Clinical relevance of thrombocytosis in primary care: a prospective cohort study of cancer incidence using English electronic medical records and cancer registry data. Br J Gen Pract. 2017; 67(659): e405-e413.

[4] Ma Y, Li G, Yu M, et al. Prognostic significance of thrombocytosis in lung cancer: a systematic review and meta-analysis. Platelets. 2021; 32(7): 919-927.

[5] Yuan Y, Zhong H, Ye L, et al. Prognostic value of pretreatment platelet counts in lung cancer: a systematic review and meta-analysis. BMC Pulm Med. 2020; 20(1): 96.

[6] Giannakeas V, Kotsopoulos J, Cheung MC, et al. Analysis of platelet count and new cancer diagnosis over a 10-year period. JAMA Netw Open. 2022; 5(1):e2141633.

[7] Davis RB, Theologides A, Kennedy BJ. Comparative studies of blood coagulation and platelet aggregation in patients with cancer and nonmalignant diseases. Ann Intern Med. 1969; 71(1): 67-80.

[8] Gasic GJ, Gasic TB, Stewart CC. Antimetastatic effects associated with platelet reduction. Proc Natl Acad Sci U S A. 1968; 61(1): 46-52.

[9] Xu XR, Yousef GM, Ni H. Cancer and platelet crosstalk: opportunities and challenges for aspirin and other antiplatelet agents. Blood. 2018; 131(16): 1777-1789.

[10] Tao DL, Yunga ST, Williams CD, McCarty OJT. Aspirin and antiplatelet treatments in cancer. Blood. 2021; 137(23): 3201-3211.

[11] Arepally GM. Heparin-induced thrombocytopenia. Blood. 2017; 129(21): 2864-2872.

[12] Hou J, Cao X, Cheng Y, Wang X. Roles of TP53 gene in the development of resistance to PI3K inhibitor resistances in CRISPR-Cas9-edited lung adenocarcinoma cells. Cell Biol Toxicol. 2020; 36(5): 481-492.

[13] Moloney JN, Cotter TG. ROS signalling in the biology of cancer. Semin Cell Dev Biol. 2018; 80: 50-64.

[14] Jamasbi E, Hamelian M, Hossain MA, Varmira K. The cell cycle, cancer development and therapy. Mol Biol Rep. 2022; 49(11): 10875-10883.

[15] Dorandish S, Atali S, Ray R, et al. Differences in the relative abundance of ProBDNF and mature BDNF in A549 and H1299 human lung cancer cell media. Int J Mol Sci. 2021; 22(13):7059.

[16] Inagaki N, Kibata K, Tamaki T, Shimizu T, Nomura S. Prognostic impact of the mean platelet volume/platelet count ratio in terms of survival in advanced non-small cell lung cancer. Lung Cancer. 2014; 83(1): 97-101.

[17] Gresele P, Malvestiti M, Momi S. Anti-platelet treatments in cancer: basic and clinical research. Thromb Res. 2018; 164(suppl 1): S106-S111.

[18] Pucci F, Rickelt S, Newton AP, et al. PF4 promotes platelet production and lung cancer growth. Cell Rep. 2016; 17(7): 1764-1772.

[19] Verbeke H, De Hertogh G, Li S, et al. Expression of angiostatic platelet factor-4var/CXCL4L1 counterbalances angiogenic impulses of vascular endothelial growth factor, interleukin-8/CXCL8, and stromal cell-derived factor 1/CXCL12 in esophageal and colorectal cancer. Hum Pathol. 2010; 41(7): 990-1001.

[20] Lavergne M, Janus-Bell E, Schaff M, Gachet C, Mangin PH. Platelet integrins in tumor metastasis: do they represent a therapeutic target? Cancers (Basel). 2017; 9(10):133.

[21] Umar S, Soni R, Durgapal SD, Soman S, Balakrishnan S. A synthetic coumarin derivative (4-flourophenylacetamide-acetyl coumarin) impedes cell cycle at G0/G1 stage, induces apoptosis, and inhibits metastasis via ROS-mediated p53 and AKT signaling pathways in A549 cells. J Biochem Mol Toxicol. 2020; 34(10):e22553.

[22] Sengelen A, Onay-Ucar E. Rosmarinic acid attenuates glioblastoma cells and spheroids' growth and EMT/stem-like state by PTEN/PI3K/AKT downregulation and ERK-induced apoptosis. Phytomedicine. 2024; 135:156060.

[23] Celik DA, Gurbuz N, Togay VA, Ozcelik N. Ochratoxin A causes cell cycle arrest in G1 and G1/S phases through p53 in HK-2 cells. Toxicon. 2020; 180: 11-17.

[24] Lam YK, Yu J, Huang H, et al. TP53 R249S mutation in hepatic organoids captures the predisposing cancer risk. Hepatology. 2023; 78(3): 727-740.

[25] Yan F, Liu C, Song D, et al. Integration of clinical phenoms and metabolomics facilitates precision medicine for lung cancer. Cell Biol Toxicol. 2024; 40(1): 25.

[26] Schlesinger M. Role of platelets and platelet receptors in cancer metastasis. J Hematol Oncol. 2018; 11(1): 125.

[27] Ruytinx P, Proost P, Struyf S. CXCL4 and CXCL4L1 in cancer. Cytokine. 2018; 109: 65-71.

[28] Amisten S. A rapid and efficient platelet purification protocol for platelet gene expression studies. Methods Mol Biol. 2012; 788: 155-172.

RIGHTS & PERMISSIONS

2025 The Author(s). Clinical and Translational Discovery published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics.