Vol 7, No 3 (2022)
Original article
Published online: 2022-09-30

open access

Page views 3793
Article views/downloads 307
Get Citation

Connect on Social Media

Connect on Social Media

Icaritin induces apoptosis and downregulates RhoA/ROCK pathway in the A549 adenocarcinoma cell line

Agnieszka Żuryń1, Adrian Krajewski1
Medical Research Journal 2022;7(3):242-248.


Introduction: Icaritin is a prenylflavonoid derivative from a traditional Chinese herb, Epimedium Genus. Icaritin exerts its anti-tumour effects in various cancers mainly by inhibiting cell proliferation, inducing cell differentiation and apoptosis, and suppressing cell migration and invasion. The study aimed to evaluate the effect of icaritin on apoptosis, cell cycle, and expression of selected cell cycle proteins: RhoA, ROCK1, and ROCK2 in the A549 cell line.

Methods: Apoptosis and the cell cycle distribution were evaluated by flow cytometry. Protein expression was measured by conducting a western blot assay and immunofluorescence staining.

Results and conclusions: The study revealed that icaritin induces apoptosis and cell cycle arrest in the G2/M phase in A549 cells. The authors showed that icaritin activates the p53 protein and downregulates Rho/ROCK pathway. Additionally, the decrease in expression of thrombospondin-1 was observed which can be part of the cytoprotective response of A549 cells on ICT treatment.

Article available in PDF format

View PDF Download PDF file


  1. Guo Y, Zhang X, Meng J. and Wang Z-Y: An anticancer agent icaritin induces sustained activation of the extracellular signal-regulated kinase (ERK) pathway and inhibits growth of breast cancer cells. Eur J Pharmacol 658: 114–122. ; 2011.
  2. Han S, Gou Y, Jin D, et al. and Dong X: Effects of Icaritin on the physiological activities of esophageal cancer stem cells. Biochem Biophys Res Commun 504: 792–796. ; 2018.
  3. Liu Y, Shi L, Liu Y, et al. Activation of PPARγ mediates icaritin-induced cell cycle arrest and apoptosis in glioblastoma multiforme. Biomed Pharmacother. 2018; 100: 358–366.
  4. Sun Li, Peng Q, Qu L, et al. Anticancer agent icaritin induces apoptosis through caspase-dependent pathways in human hepatocellular carcinoma cells. Mol Med Rep. 2015; 11(4): 3094–3100.
  5. Wu T, Wang S, Wu J, et al. Icaritin induces lytic cytotoxicity in extranodal NK/T-cell lymphoma. J Exp Clin Cancer Res. 2015; 34: 17.
  6. Zheng Q, Liu WW, Li B, et al. Anticancer effect of icaritin on human lung cancer cells through inducing S phase cell cycle arrest and apoptosis. J Huazhong Univ Sci Technolog Med Sci. 2014; 34(4): 497–503.
  7. Zhu S, Wang Z, Li Z, et al. Icaritin suppresses multiple myeloma, by inhibiting IL-6/JAK2/STAT3. Oncotarget. 2015; 6(12): 10460–10472.
  8. Matsuoka T, Yashiro M. Rho/ROCK signaling in motility and metastasis of gastric cancer. World J Gastroenterol. 2014; 20(38): 13756–13766.
  9. Marei H, Malliri A. Rac1 in human diseases: The therapeutic potential of targeting Rac1 signaling regulatory mechanisms. Small GTPases. 2017; 8(3): 139–163.
  10. Hall A. Rho family GTPases. Biochem Soc Trans 40: 1378. 1382; 2012.
  11. Orgaz JL, Herraiz C, Sanz-Moreno V. Rho GTPases modulate malignant transformation of tumor cells. Small GTPases. 2014; 5: e29019.
  12. Sadok A, Marshall CJ. Rho GTPases: masters of cell migration. Small GTPases. 2014; 5: e29710.
  13. Mack NA, Whalley HJ, Castillo-Lluva S, et al. The diverse roles of Rac signaling in tumorigenesis. Cell Cycle. 2011; 10(10): 1571–1581.
  14. Porter AP, Papaioannou A, Malliri A. Deregulation of Rho GTPases in cancer. Small GTPases. 2016; 7(3): 123–138.
  15. Żuryń A, Krajewski A. Klimaszewska‑Wiśniewska A, Grzanka A and Grzanka D: Expression of cyclin B1, D1 and K in non‑small cell lung cancer H1299 cells following treatment with sulforaphane. Oncol Rep. ; 2018.
  16. Zhao X, Lin Y, Jiang B, et al. and Zeng J: Icaritin inhibits lung cancer-induced osteoclastogenesis by suppressing the expression of IL-6 and TNF-a and through AMPK/mTOR signaling pathway. Anticancer Drugs 31: 1004. 1011; 2020.
  17. Zhou C, Gu J, Zhang G, et al. Chen M-B and Xu D: AMPK-autophagy inhibition sensitizes icaritin-induced anti-colorectal cancer cell activity. Oncotarget 8: 14736–1. 4747; 2017.
  18. Huang C, Li Z, Zhu J, et al. Systems pharmacology dissection of targeting tumor microenvironment to enhance cytotoxic T lymphocyte responses in lung cancer. Aging (Albany NY). 2021; 13(2): 2912–2940.
  19. Chen X, Song L, Hou Y. and Li F: Reactive oxygen species induced by icaritin promote DNA strand breaks and apoptosis in human cervical cancer cells. Oncol Rep. ; 2018.
  20. Hu C, Zhou H, Liu Y, et al. ROCK1 promotes migration and invasion of non‑small‑cell lung cancer cells through the PTEN/PI3K/FAK pathway. Int J Oncol. 2019; 55(4): 833–844.
  21. Vigil D, Kim TY, Plachco A, et al. ROCK1 and ROCK2 are required for non-small cell lung cancer anchorage-independent growth and invasion. Cancer Res. 2012; 72(20): 5338–5347.
  22. Zhang Z, Nong Li, Chen M, et al. Baicalein suppresses vasculogenic mimicry through inhibiting RhoA/ROCK expression in lung cancer A549 cell line. Acta Biochim Biophys Sin (Shanghai). 2020; 52(9): 1007–1015.
  23. Chen Z, Le H, Zhang Y, et al. and Li W: Microvessel Density and Expression of Thrombospondin-1 in Non-small Cell Lung Cancer and Their Correlation with Clinicopathological Features. J Int Med Res 37: 551–556. ; 2009.
  24. Rouanne M, Adam J, Goubar A, et al. Osteopontin and thrombospondin-1 play opposite roles in promoting tumor aggressiveness of primary resected non-small cell lung cancer. BMC Cancer. 2016; 16: 483.
  25. Daubon T, Léon C, Clarke K, et al. Deciphering the complex role of thrombospondin-1 in glioblastoma development. Nat Commun. 2019; 10(1): 1146.
  26. Guillon J, Petit C, Moreau M, et al. Regulation of senescence escape by TSP1 and CD47 following chemotherapy treatment. Cell Death Dis. 2019; 10(3): 199.
  27. Shan SW, Do CW, Lam TC, et al. Thrombospondin-1 mediates Rho-kinase inhibitor-induced increase in outflow-facility. J Cell Physiol. 2021; 236(12): 8226–8238.
  28. Borsotti P, Ghilardi C, Ostano P, et al. Thrombospondin-1 is part of a Slug-independent motility and metastatic program in cutaneous melanoma, in association with VEGFR-1 and FGF-2. Pigment Cell Melanoma Res. 2015; 28(1): 73–81.