International Journal of Clinical Oncology

, Volume 23, Issue 2, pp 321–328 | Cite as

Combined detection of Twist1, Snail1 and squamous cell carcinoma antigen for the prognostic evaluation of invasion and metastasis in cervical squamous cell carcinoma

  • Huilun Yang
  • Haiyang Hu
  • Yanling Gou
  • Yuhong Hu
  • Hui Li
  • Hongwei Zhao
  • Beidi Wang
  • Peiling Li
  • Zongfeng ZhangEmail author
Original Article



Cervical cancer is one of the most common malignant tumours of the female reproductive system, ranking second only to breast cancer in morbidity worldwide. Essential features of the progression of cervical cancer are invasion and metastasis, which are closely related to disease prognosis and mortality rate. At the present time there is no effective method to evaluate cancer invasion and metastasis before surgery. Here we report our study on molecular changes in biopsy tissue for the prognostic evaluation of cancer invasion and metastasis.

Patients and methods

Expression of the epithelial–mesenchymal transition-inducing transcription factors Twist1 and Snail1 was detected by immunohistochemistry in 32 normal, 36 low-grade squamous intraepithelial neoplasia (LSIL), 54 high-grade squamous intraepithelial neoplasia (HSIL) and 320 cervical squamous cell carcinoma (CSCC) samples. The correlation between the expression of Twist1, Snail1 and squamous cell carcinoma antigen (SCCA) in CSCC tissues and clinical pathology results was evaluated. A transwell migration and invasion assay was used to explore the roles of Twist1 and Snail1 in the invasion of cancer cells. Lymph node metastasis and lymphovascular space invasion (LVSI) rates for the following groups were analysed: SCCA(+) group, Twist1(+) group, Snail1(+) group, Twist1(+)Snail1(+)group, Twist1(+)SCCA(+)group, Snail1(+)SCCA(+)group and Twist1(+)Snail1(+)SCCA(+) group.


The expression of Twist1 and Snail1 was significantly upregulated in HSIL and CSCC (p < 0.05). Twist1 and Snail1 expression levels were associated with LVSI, lymph node metastasis and histological grade (p < 0.05) but not with age or FIGO stage (p > 0.05). The expression of SCCA was associated with LVSI, lymph node metastasis, FIGO stage and histological grade (p < 0.05) but not with age (p > 0.05). Twist1 was an independent factor contributing to the invasion ability of cervical cancer cells. In addition, the positive rate of lymph node metastasis and LVSI was higher in the Twist1(+)Snail1(+)SCCA(+) group than in the SCCA(+) group, Twist1(+) group and Snail1(+) group, respectively (p < 0.05).


Combined detection of Twist1 and Snail1 in SCCA-positive biopsy specimens may be a potential method for evaluating the invasion and metastasis of CSCC prior to surgery.


Twist1 Snail1 Squamous cell carcinoma antigen Cervical squamous cell carcinoma 


Compliance with ethical standards

Informed consent

Informed consent was obtained from each participants included in the study.

Conflicts of interest

The authors declare that they have no conflict of interest.


This study was funded by National Natural Science Foundation of China (81671430).

Ethical standards

Ethical approval was given by the medical ethics committee of Second Affiliated Hospital of Harbin Medical University. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. And this article does not contain any studies with animals performed by any of the authors.


  1. 1.
    Qi W, Ai Z, Liu X et al (2012) Expression of transient receptor potential channel 6 in cervical cancer. Oncol Targets Ther 5:171–176Google Scholar
  2. 2.
    Zhou X, Xu CJ, Wang JX et al (2015) Metastasis-associated in colon cancer-1 associates with poor prognosis and promotes cell invasion and angiogenesis in human cervical cancer. Int J Gynecol Cancer 25:1353–1363CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    De CB, Berx G (2013) Regulatory networks defining EMT during cancer initiation and progression. Nat Rev Cancer 13:97–110CrossRefGoogle Scholar
  4. 4.
    Ansieau S, Bastid J, Doreau A et al (2008) Induction of EMT by twist proteins as a collateral effect of tumor-promoting inactivation of premature senescence. Cancer Cell 14:79–89CrossRefPubMedGoogle Scholar
  5. 5.
    Xu Y, Qin L, Sun T et al (2017) Twist1 promotes breast cancer invasion and metastasis by silencing Foxa1 expression. Oncogene 36:1157–1166CrossRefPubMedGoogle Scholar
  6. 6.
    Chang TM, Hung WC (2012) Transcriptional repression of TWIST1, gene by Prospero-related homeobox 1 inhibits invasiveness of hepatocellular carcinoma cells. FEBS Lett 586:3746–3752CrossRefPubMedGoogle Scholar
  7. 7.
    Zhu X, Shen H, Yin X et al (2016) Mir-186 regulation of twist1 and ovarian cancer sensitivity to cisplatin. Oncogene 35:323–332CrossRefPubMedGoogle Scholar
  8. 8.
    Kaufhold S, Bonavida B (2014) Central role of Snail1 in the regulation of EMT and resistance in cancer: a target for therapeutic intervention. J Exp Clin Cancer Res 33:62CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Muenst S, Däster S, Obermann EC et al (2013) Nuclear expression of snail is an independent negative prognostic factor in human breast cancer. Dis Markers 35:337–344CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Wang YL, Zhao XM, Shuai ZF et al (2014) Snail promotes epithelial-mesenchymal transition and invasiveness in human ovarian cancer cells. Int J Clin Exp Med 8:7388–7393Google Scholar
  11. 11.
    Brzozowa M, Michalski M, Wyrobiec G et al (2014) The role of Snail1 transcription factor in colorectal cancer progression and metastasis. Contemp Oncol 4:265–270Google Scholar
  12. 12.
    Salvatici M, Achilarre MT, Sandri MT et al (2016) Squamous cell carcinoma antigen (SCC-Ag) during follow-up of cervical cancer patients: role in the early diagnosis of recurrence. Gynecol Oncol 142:115–119CrossRefPubMedGoogle Scholar
  13. 13.
    Chang SJ, Bristow RE, Ryu HS (2012) A model for prediction of parametrial involvement and feasibility of less radical resection of parametrium in patients with FIGO stage IB1 cervical cancer. Gynecol Oncol 126:82–86CrossRefPubMedGoogle Scholar
  14. 14.
    Kajiyama H, Shibata K, Umezu T et al (2013) Expression of Twist enhances risk of poor oncologic outcome in patients with stage Ib to II cervical carcinoma with lymphovascular space involvement. Hum Pathol 44:181–188CrossRefPubMedGoogle Scholar
  15. 15.
    Shibata K, Kajiyama H, Ino K et al (2008) Twist expression in patients with cervical cancer is associated with poor disease outcome. Ann Oncol 19:81–85CrossRefPubMedGoogle Scholar
  16. 16.
    Zhilan C, Shuang L, Kecheng H et al (2013) The nuclear protein expression levels of SNAI1 and ZEB1 are involved in the progression and lymph node metastasis of cervical cancer via the epithelial-mesenchymal transition pathway. Hum Pathol 44:2097–2105CrossRefGoogle Scholar
  17. 17.
    Li Y, Wang W, Wang W et al (2012) Correlation of TWIST2 up-regulation and epithelial-mesenchymal transition during tumorigenesis and progression of cervical carcinoma. Gynecol Oncol 124:112–118CrossRefPubMedGoogle Scholar
  18. 18.
    Liu Y, Qian W, Zhang J et al (2015) The indicative function of Twist2 and E-cadherin in HPV oncogene-induced epithelial-mesenchymal transition of cervical cancer cells. Oncol Rep 33:639–650CrossRefPubMedGoogle Scholar
  19. 19.
    Chung HH, Kang SB, Cho JY et al (2007) Can preoperative MRI accurately evaluate nodal and parametrial invasion in early stage cervical cancer? Jpn J Clin Oncol 37:370–375CrossRefPubMedGoogle Scholar
  20. 20.
    Bipat S, Glas AS, Dvj Van et al (2003) Computed tomography and magnetic resonance imaging in staging of uterine cervical carcinoma: a systematic review. Gynecol Oncol 91:59–66CrossRefPubMedGoogle Scholar
  21. 21.
    Choi HJ, Ju W, Myung SK (2010) Diagnostic performance of computer tomography, magnetic resonance imaging, and positron emission tomography or positron emission tomography/computer tomography for detection of metastatic lymph nodes in patients with cervical cancer: meta-analysis. Cancer Sci 101:1471–1479CrossRefPubMedGoogle Scholar
  22. 22.
    Puisieux A, Brabletz T, Caramel J (2014) Oncogenic roles of EMT-inducing transcription factors. Nat Cell Biol 16:488–494CrossRefPubMedGoogle Scholar
  23. 23.
    Cheng GZ, Chan J, Wang Q et al (2007) Twist transcriptionally up-regulates AKT2 in breast cancer cells leading to increased migration, invasion, and resistance to paclitaxel. Cancer Res 67:1979–1987CrossRefPubMedGoogle Scholar
  24. 24.
    Wang WS, Yang XS, Xia M et al (2012) Silencing of twist expression by RNA interference suppresses epithelial-mesenchymal transition, invasion, and metastasis of ovarian cancer. Asian Pac J Cancer Prev 13:4435–4439CrossRefPubMedGoogle Scholar
  25. 25.
    Fan XJ, Wan XB, Yang ZL et al (2013) Snail promotes lymph node metastasis and twist enhances tumor deposit formation through epithelial-mesenchymal transition in colorectal cancer. Hum Pathol 44:173–180CrossRefPubMedGoogle Scholar
  26. 26.
    Herreros AGD, Peiró S, Nassour M et al (2010) Snail family regulation and epithelial mesenchymal transitions in breast cancer progression. J Mammary Gland Biol Neoplasia 15:135–147CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Kurrey NK, Amit K, Bapat SA (2005) Snail and Slug are major determinants of ovarian cancer invasiveness at the transcription level. Gynecol Oncol 97:155–165CrossRefPubMedGoogle Scholar
  28. 28.
    Lekskul N, Charakorn C, Lertkhachonsuk AA et al (2015) The level of squamous cell carcinoma antigen and lymph node metastasis in locally advanced cervical cancer. Asian Pac J Cancer Prev 16:4719–4722CrossRefPubMedGoogle Scholar
  29. 29.
    Water TJ, Berkhof J (2009) SCC-Ag, lymph node metastases and sentinel node procedure in early stage squamous cell cervical cancer. Gynecol Oncol 112:119–125CrossRefPubMedGoogle Scholar
  30. 30.
    Katagiri C, Nakanishi J, Kadoya K (2006) Serpin squamous cell carcinoma antigen inhibits UV-induced apoptosis via suppression of c-JUN NH2-terminal kinase. J Cell Biol 172:983–990CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Suminami Y, Nagashima S, Murakami A et al (2001) Suppression of a squamous cell carcinoma (SCC)-related serpin, SCC antigen, inhibits tumor growth with increased intratumor infiltration of natural killer cells. Cancer Res 61:1776–1780PubMedGoogle Scholar
  32. 32.
    Quarta S, Vidalino L, Turato C et al (2010) SERPINB3 induces epithelial-mesenchymal transition. J Pathol 221:343–356CrossRefPubMedGoogle Scholar
  33. 33.
    Zheng X, Xing S, Liu XM et al (2014) Establishment of using serum YKL-40 and SCCA in combination for the diagnosis of patients with esophageal squamous cell carcinoma. BMC Cancer 14:490CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Japan Society of Clinical Oncology 2017

Authors and Affiliations

  • Huilun Yang
    • 1
    • 2
  • Haiyang Hu
    • 1
    • 3
  • Yanling Gou
    • 1
  • Yuhong Hu
    • 4
  • Hui Li
    • 1
  • Hongwei Zhao
    • 1
  • Beidi Wang
    • 1
  • Peiling Li
    • 1
  • Zongfeng Zhang
    • 1
    Email author
  1. 1.Department of Obstetrics and GynecologySecond Affiliated Hospital of Harbin Medical UniversityHarbinChina
  2. 2.Department of Obstetrics and GynecologyThe First People’s Hospital of YangzhouYangzhouChina
  3. 3.Department of Obstetrics and GynecologyAffiliated Hospital of Jining Medical UniversityJiningChina
  4. 4.Department of Obstetrics and GynecologyThe First Hospital Affiliated to Jiamusi UniversityJiamusiChina

Personalised recommendations