Short-hairpin RNA-mediated suppression of cortactin may inhibit the migration and invasion abilities of endometrial cancer cells by reducing lamellipodia

Document Type : Original Article


1 Department of Gynecology and Obstetrics, The First Affiliated Hospital of Guangxi Medical University, Nanning, China

2 Department of Gynecology and Obstetrics, University of the Chinese Academy of Sciences, Shenzhen Hospital, Shenzhen, China

3 Key Laboratory of Early Prevention and Treatment for Regional High-Frequency Tumor (Guangxi Medical University), Ministry of Education, Nanning, China


Objective(s): The prognosis of endometrial cancer (EC) is significantly affected by tumor infiltration and metastasis. Cortactin (CTTN) regulates infiltration and metastasis in other tumors. Studies on the role and mechanism of CTTN in EC are limited and further studies are needed.
Materials and Methods: Quantitative PCR and immunohistochemistry were used to detect Ras-associated C3 botulinum toxin substrate 1 (Rac1) and CTTN in EC and normal tissues. The relationship between the expression of these two genes and their prognostic factors was analyzed. A CTTN-RNAi lentiviral system was constructed and transfected into EC cells. Migration and invasion were evaluated by scratch assay, transwell migration, and invasion assays. Pseudopodia formation was observed by immunofluorescence staining. Western blotting was performed to detect the expression of Rac1.
Results: The expression levels of Rac1 and CTTN in EC tissues were significantly higher than those in normal tissues. In the EC group, Rac1 and CTTN levels were correlated. The protein expression levels of Rac1 and CTTN were related to myometrial invasion and stage. After CTTN knockdown, the migration rate, invasiveness, and migratory ability of EC cells decreased significantly. Lamellipodia was observed to disappear with the appearance of blebs. Rac1 protein expression was decreased after CTTN knockdown.
Conclusion: CTTN may promote the invasion and migration of EC by lamellipodia. This effect may be related to the regulation of Rac1 by CTTN.


Main Subjects

1. Urick ME, Bell DW. Clinical actionability of molecular targets in endometrial cancer. Nat Rev Cancer 2019;19:510-521.
2. Jonusiene V, Sasnauskiene A. Notch and endometrial cancer. Adv Exp Med Biol 2021;1287:47-57. 
3. Hamilton CA, Pothuri B, Arend RC, Backes FJ, Gehrig PA, Soliman PT, et al. Endometrial cancer: A society of gynecologic oncology evidence-based review and recommendations. Gynecol Oncol 2021;160:817-826. 
4. Colombo N, Creutzberg C, Amant F, Bosse T, González-Martín A, Ledermann J, et al. ESMO-ESGO-ESTRO consensus conference on endometrial cancer: Diagnosis, treatment and follow-up. Int J Gynecol Cancer 2016;26:2-30. 
5. Chen J, Gao F, Liu N. L1CAM promotes epithelial to mesenchymal transition and formation of cancer initiating cells in human endometrial cancer. Exp Ther Med 2018;15:2792-2797. 
6. Wu H, Parsons JT. Cortactin, an 80/85-kilodalton pp60src substrate, is a filamentous actin-binding protein enriched in the cell cortex. J Cell Biol 1993;120:1417-1426.
7. Svitkina TM. Actin cell cortex: Structure and molecular organization. Trends Cell Biol 2020;30:556-565. 
8. Schnoor M, Stradal TE, Rottner K. Cortactin: Cell functions of a multifaceted actin-binding protein. Trends Cell Biol 2018;28:79-98. 
9. Yin M, Ma W, An L. Cortactin in cancer cell migration and invasion. Oncotarget. 2017;8:88232-88243. 
10. Ramos-García P, González-Moles MÁ, González-Ruiz L, Ayén Á, Ruiz-Ávila I, Navarro-Triviño FJ, et al. An update of knowledge on cortactin as a metastatic driver and potential therapeutic target in oral squamous cell carcinoma. Oral Dis 2019;25:949-971.
11. Li A, Zhang L, Zhang X, Jin W, Ren Y. Expression and clinical significance of cortactin protein in ovarian neoplasms. Clin Transl Oncol 2016;18:220-227. 
12. Watkins RJ, Imruetaicharoenchoke W, Read ML, Sharma N, Poole VL, Gentilin E, et al. Pro-invasive effect of proto-oncogene pbf is modulated by an interaction with cortactin. J Clin Endocrinol Metab 2016;101:4551-4563. 
13. Ji J, Feng X, Shi M, Cai Q, Yu Y, Zhu Z, et al. Rac1 is correlated with aggressiveness and a potential therapeutic target for gastric cancer. Int J Oncol 2015;46:1343-1353. 
14. Bao Y, Guo HH, Lu Y, Feng W, Sun X, Tang C, et al. Blocking hepatic metastases of colon cancer cells using an shRNA against Rac1 delivered by activatable cell-penetrating peptide. Oncotarget 2016;7:77183-77195. 
15. Melzer C, Hass R, von der Ohe J, Lehnert H, Ungefroren H. The role of TGF-β and its crosstalk with RAC1/RAC1b signaling in breast and pancreas carcinoma. Cell Commun Signal. 2017;15:1-11. 
16. De P, Aske JC, Dey N. RAC1 takes the lead in solid tumors. Cells 2019;8:1-17. 
17. Ji R, Zhu XJ, Wang ZR, Huang LQ. Cortactin in epithelial-mesenchymal transition. Front Cell Dev Biol. 2020 Oct 20;8:585619. 
18. Head JA, Jiang D, Li M, Zorn LJ, Schaefer EM, Parsons JT, et al. Cortactin tyrosine phosphorylation requires Rac1 activity and association with the cortical actin cytoskeleton. Mol Biol Cell 2003;14:3216-3229. 
19. Song JX, Kong LP, Du ZL. Expression of ras-related C3 botulinum toxin substrate 1 in endometrial carcinoma tissue and its clinical significance. J Precision Medicine 2017;32:636.
20. Hu M, Li Z, Qiu J, Zhang R, Feng J, Hu G, et al. CKS2 (CDC28 protein kinase regulatory subunit 2) is a prognostic biomarker in lower grade glioma: a study based on bioinformatic analysis and immunohistochemistry. Bioengineered 2021;12:5996-6009. 
21. Fromowitz FB, Viola MV, Chao S,Oravez S, Mishriki Y, Finkel G, et al. Ras p21 expression in the progression of breast cancer. Hum Pathol 1987;18:1268-1275. 
22. Clark EA, King WG, Brugge JS, Symons M, Hynes RO. Integrin-mediated signals regulated by members of the rho family of GTPases. J Cell Biol 1998;142:573-586.
23. Hall A. Rho GTPases and the actin cytoskeleton. Science 1998;279:509-514. 
24. Ridley AJ, Schwartz MA, Burridge K, Firtel RA, Ginsberg MH, Borisy G, et al. Cell migration: integrating signals from front to back. Science 2003;302:1704-1709. 
25. Rottner K, Schaks M. Assembling actin filaments for protrusion. Curr Opin Cell Biol 2019;56:53-63. 
26. Blanchoin L, Boujemaa-Paterski R, Sykes C, Plastino J. Actin dynamics, architecture, and mechanics in cell motility. Physiol Rev 2014;94:235-263.
27. Small JV, Stradal T, Vignal E, Rottner K. The lamellipodium: where motility begins. Trends Cell Biol 2002;12:112-120. 
28. Beli P, Mascheroni D, Xu D,Innocenti M. WAVE and Arp2/3 jointly inhibit filopodium formation by entering into a complex with mDia2. Nat Cell Biol 2008;10:849-857.
29. Borm B, Requardt RP, Herzog V, Kirfel G. Membrane ruffles in cell migration: indicators of inefficient lamellipodia adhesion and compartments of actin filament reorganization. Exp Cell Res 2005;302:83-95. 
30. Hogue MJ. The effect of hypotonic and hypertonic solutions on fibroblasts of the embryonic chick heart in vitro. J Exp Med 1919;30:617-648. 
31. Charras GT. A short history of blebbing. J Microsc 2008;231:466-478.
32. Charras G, Paluch E. Blebs lead the way: How to migrate without lamellipodia. Nat Rev Mol Cell Biol 2008;9:730-736.  
33. Lawson CD, Ridley AJ. Rho GTPase signaling complexes in cell migration and invasion. J Cell Biol 2018;217:447-457. 
34. Bergert M, Chandradoss SD, Desai RA, Paluch E. Cell mechanics control rapid transitions between blebs and lamellipodia during migration. Proc Natl Acad Sci U S A 2012;109:14434-14439.
35. Chikina AS, Svitkina TM, Alexandrova AY. Time-resolved ultrastructure of the cortical actin cytoskeleton in dynamic membrane blebs. J Cell Biol 2019;218:445-454. 
36. Paluch EK, Raz E. The role and regulation of blebs in cell migration. Curr Opin Cell Biol 2013;25:582-590. 
37. Hudson LG, Gillette JM, Kang H, Rivera MR, Wandinger-Ness A. Ovarian tumor microenvironment signaling: convergence on the Rac1 GTPase. Cancers (Basel) 2018;10:1-26. 
38. Zhou K, Rao J, Zhou ZH, Yao XH, Wu F, Yang J, et al. RAC1-GTP promotes epithelial-mesenchymal transition and invasion of colorectal cancer by activation of STAT3. Lab Invest 2018;98:989-998. 
39. Hohmann T, Dehghani F. The Ccytoskeleton-A complex interacting meshwork. Cells. 2019;8:1-55. 
40. Lozano E, Betson M, Braga VM. Tumor progression: Small GTPases and loss of cell-cell adhesion. Bioessays 2003;25:452-463. 
41. Wang Y, Hu L, Ji P, Teng F, Tian W, Liu Y, et al. MIIP remodels Rac1-mediated cytoskeleton structure in suppression of endometrial cancer metastasis. J Hematol Oncol 2016;9:1-10. 
42. Rane CK, Minden A. P21 activated kinases: structure, regulation, and functions. Small GTPases 2014;5:e28003. 
43. Lai FP, Szczodrak M, Oelkers JM, Ladwein M, Acconcia F, Benesch S, et al. Cortactin promotes migration and platelet-derived growth factor-induced actin reorganization by signaling to Rho-GTPases. Mol Biol Cell 2009;20:3209-3223. 
44. Lim Lam VK, Hin Wong JY, Chew SY, Chan BP. Rac1-GTPase regulates compression-induced actin protrusions (CAPs) of mesenchymal stem cells in 3D collagen micro-tissues. Biomaterials 2021;274:120829. 
45. Krause M, Gautreau A. Steering cell migration: Lamellipodium dynamics and the regulation of directional persistence. Nat Rev Mol Cell Biol 2014;15:577-590.