Expression of Transcription Factor FOXC2 in Cervical Cancer and Effects of Silencing on Cervical Cancer Cell Proliferation

  • Zheng, Chun-Hua (Gynecology, The First Hospital of Jilin University) ;
  • Quan, Yuan (Gynecology, The First Hospital of Jilin University) ;
  • Li, Yi-Yang (Gynecology, The First Hospital of Jilin University) ;
  • Deng, Wei-Guo (Public Health, Jilin University) ;
  • Shao, Wen-Jing (Gynecology, The First Hospital of Jilin University) ;
  • Fu, Yan (Gynecology, The First Hospital of Jilin University)
  • Published : 2014.02.28


Objective: Forkhead box C2 (FOXC2) is a member of the winged helix/forkhead box (Fox) family of transcription factors. It has been suggested to regulate tumor vasculature, growth, invasion and metastasis, although it has not been studied in cervical cancer. Here, we analyzed FOXC2 expression in cervical tissues corresponding to different stages of cervical cancer development and examined its correlation with clinicopathological characteristics. In addition, we examined the effects of targeting FOXC2 on the biological behavior of human cervical cancer cells. Methods: The expression of FOXC2 in normal human cervix, CIN I-III and cervical cancer was examined by immunohistochemistry and compared among the three groups and between cervical cancers with different pathological subtypes. Endogenous expression of FOXC2 was transiently knocked down in human Hela and SiHa cervical cells by siRNA, and cell viability and migration were examined by scratch and CCK8 assays, respectively. Results: In normal cervical tissue the frequency of positive staining was 25% (10/40 cases), with a staining intensity (PI) of $0.297{\pm}0.520$, in CIN was 65% (26/40cases), with a PI of $3.00{\pm}3.29$, and in cancer was 91.8% (68/74 cases), with a PI of $5.568 {\pm}3.449$. The frequency was 100% in adenocarcinoma (5/5 cases) and 91.3% in SCCs (63/69 cases). The FOXC2 positive expression rate was 88.5% in patients with cervical SCC stage I and 100% in stage II, showing significant differences compared with normal cervix and CIN. With age, pathologic differentiation degree and tumor size, FOXC2 expression showed no significant variation. On transient transfection of Hela and SiHa cells, FOXC2-siRNA inhibition rates were 76.2% and 75.7%; CCK8 results showed reduced proliferation and relative migration (in Hela cells from $64.5{\pm}3.16$ to $49.5{\pm}9.24$ and in SiHa cells from $60.1{\pm}3.05$ to $44.3{\pm}3.98$) (P < 0.05). Conclusion: FOXC2 gene expression increases with malignancy, especially with blood vessel hyperplasia and invasion degree. Targeted silencing was associated with reduced cell proliferation as well as invasion potential.


  1. VanTrappen PO, Ryan A, Carroll M, Lecoeur C (2002). A model for coexpression pattern analysis of genes implicated in angiogenesis and tumour cell invasion in cervical cancer. Br J Cancer, 87, 537-44.
  2. Vreeburg M, Heitink MV, Damstra RJ, et al (2008). Lymphedemadistichiasis syndrome: a distinct type of primary lymphedema caused by mutations in the FOXC2 gene. Int J Dermatol, 47, 52-5.
  3. Wang WS, Yu SL, Yang XS, Chang SD, Hou JQ (2013) . Expression and Significance of Twist and E-cadherin in Ovarian Cancer Tissues. Asian Pac J Cancer Prev, 14, 669-72.
  4. Yang SH, Cheng HH, Wang ZH (2008). LYVE-1 and D2-40 in the study of early carcinoma and precancerous lesions of cervical squamous cell lymphatic microvessel in China detection. J Histochem Cytochem, 17, 588-92.
  5. Yu YH, Chen HA, Chen PS, et al (2013). MiR-520h-mediated FOXC2 regulation is critical for inhibition of lung cancer progression by resveratrol. Oncogene, 32, 431-43.
  6. Pichler A, Zelcer N, Prior JL, et al (2005). In vivo RNA interference-mediated ablation of MDR1 P-glycoprotein. Clin Cancer Res, 11, 4487-94.
  7. Mani SA, Yang J, Brooks M, et al (2007). Mesenchyme Forkhead 1 (FOXC2) plays a key role in metastasis and is associated with aggressive basal-like breast cancers. Proc Natl Acad Sci USA, 104, 10069-74.
  8. Naohiro N, Koshi M, Takehiko Y, et al (2011). FOXC2 is a Novel Prognostic Factor in Human Esophageal SCC.Ann Surg Oncol, 18, 535-42.
  9. Nishida N, Mimori K, Yokobori T, et al (2011). FOXC2 is a novel prognostic factor in human esophageal SCC. Ann Surg Oncol, 18, 535-42.
  10. Petrova TV, Karpanen T, Norrmen C, et al (2004). Defective valves and abnormal mural cell recruitment underlie lymphatic vascular failure in lymphedema distichiasia. Nat Med, 10, 974-81.
  11. Putral LN, Bywater MJ, Gu W, et al (2005). RNA interference against human papillomavirus oncogenes in cervical cancer cells results in increased sensitivity to cisplatin. Mol Pharmacol, 68, 1311-9.
  12. Roxanis I (2013). Occurrence and significance of epithelialmesenchymal transition in breast cancer. J Clin Pathol, 66, 517-21.
  13. Sano H, Leboeuf JP, Novitskiy SV, et al (2010). The FOXC2 transcription factor regulates tumor angiogenesis. Biochem Biophys Res Commun, 392, 201-6.
  14. Sendurai A.Mani, Jing Y, Mary B, et al (2007). Mesenchyme Forkhead 1 (FOXC2) plays a key role in metastasis and is associated with aggressive basal-like breast cancers.Proc Natl Acad Sci USA, 104, 10069-74.
  15. Sima N, Wang W, Kong D, et al (2008). RNA interference against HPV16E7 oncogene leads to viral E6 and E7 suppression in cervical cancer cells and apoptosis via upregulation of Rb and P53.Apoptosis, 13, 273-81.
  16. Kreis NN, Sanhaji M, Kramer A, et al (2010). Restoration of the tumor suppressor p53 by down regulating cyclin B1 in human papilloma virus 16 /18 infected cancer cells. Oncogene, 29, 5591-603.
  17. Hayashi H, Kume T (2008). Foxc transcription factors directly regulate Dll4 and Hey2 expression by interacting with the VEGF-notch signaling pathways in endothelial cells.Plos One, 3, 2401.
  18. Itoh M, Nakagawa H (2013). A novel complex insertion-deletion mutation in the FOXC2 gene in a Japanese patient with lymphedema-distichiasis Syndrome. Eur J Dermatol, 23, 411-13.
  19. Jiang XY, Liu WK, Zhang ZX, et al (2009). According to the HPV16 E7 gene RNA interference on the biological characteristics of CaSKi cell modern. Oncology, 17, 1029-31.
  20. Kreis NN, Sanhaji M, Kramer A, et al (2010). Restoration of the tumorsuppressor p53 by downregulating cyclin B1 in human papilloma-virus 16 /18-infected cancer cells.Oncogene, 29, 5591-603.
  21. Kriederman BM, Myloyde TL, Witte MH, et al (2003). FOXC2 haploinsufficient mice are a model for human autosomal dominant lymphedema-distichiasis syndrome. Hum Mol Genet, 12, 1179-85.
  22. Kuhnert F, Jessica RK, Thurston G (2011). Dll4-notch signaling as a therapeutic target in tumor angiogenesis. Vascular Cell, 3, 1-8.
  23. Kume T (2008). FOXC2 transcription factor: a newly described regulator of angiogenesis. Trends Cardiovasc Med, 18, 224-8.
  24. Kume T (2012). The role of FOXC2 transcription factor in tumor angiogenesis. J Oncol, 2012, 204593.
  25. Lee MY, Shen MR (2012). Epithelial-mesenchymal transition in cervical carcinoma.Am J Transl Res, 4, 1-13.
  26. Dellinger MT, Thome K, Bernas MJ, Erickson RP, Witte MH (2008). Novel FOXC2 missense mutation identified in patient with lymphedema-distichiasis syndrome and review. Lymphology, 41, 98-102.
  27. Forouzanfar MH, Foreman KJ, Delossantos AM, et al (2011). Breast and cervical cancer in 187 countries between 1980 and 2010: a systematic analysis. Lance Oct, 1461-84.

Cited by

  1. Comprehensive Expression Analysis Suggests Functional Overlapping of Human FOX Transcription Factors in Cancer vol.15, pp.23, 2015,
  2. High Expression of Forkhead Box Protein C2 is Related to Poor Prognosis in Human Gliomas vol.15, pp.24, 2015,
  3. 'Drawing' a Molecular Portrait of CIN and Cervical Cancer: a Review of Genome-Wide Molecular Profiling Data vol.16, pp.11, 2015,
  4. FOXC2 is up-regulated in pancreatic ductal adenocarcinoma and promotes the growth and migration of cancer cells vol.37, pp.7, 2016,