DOI QR코드

DOI QR Code

Silencing of Twist Expression by RNA Interference Suppresses Epithelial-mesenchymal Transition, Invasion, and Metastasis of Ovarian Cancer

  • Wang, Wen-Shuang ;
  • Yang, Xing-Sheng ;
  • Xia, Min ;
  • Jiang, Hai-Yang ;
  • Hou, Jian-Qing
  • Published : 2012.09.30

Abstract

Purpose: This study aimed to explore the role of the Twist gene in the epithelial-mesenchymal transition of ovarian cancer. Methods: An RNA interference plasmid expressing a small interfering RNA (siRNA)-targeting Twist (Twist siRNA vector) was designed, constructed, and transfected into the human ovarian cancer cell line A2780. Transfection efficiency was assessed under a fluorescence microscope. Changes in the expression of Twist mRNA in A2780 after transfection with the pGenesil Twist shRNA plasmid were analyzed through RT-PCR. MTT assays and adhesion experiments were applied to determine changes in proliferation and adhesion ability of A2870 after transfection with the Twist shRNA plasmid. Changes in the expression of the E-cadherin and N-cadherin proteins in A2780 after transfection with the Twist shRNA plasmid were analyzed using Western blotting. Result: The restructuring plasmid pGenesil-Twist shRNA was constructed successfully. After 48 h of culture, 80% of the cells expressed high-intensity GFP fluorescence and stability. The expression of Twist decreased significantly after the transfection of the Twist shRNA plasmid (P<0.05). Proliferation of the transfected Twist shRNA cells showed no difference with that of the A2780-nontransfection or A2780-si-control groups (P>0.05) but the adhesion ability of A2780 decreased dramatically (P<0.05). Expression of the E-cadherin protein increased, whereas that of the N-cadherin protein decreased compared with that in the A2780-nontransfection or A2780-si-control groups (P<0.05). Conclusion: Twist is essential for epithelial-mesenchymal transition, invasion, and metastasis of ovarian cancer.

Keywords

Twist;RNAi;ovarian cancer;epithelial methenchymal transition

References

  1. BoyerB, Vall S, Edme N (2000). Indution and regulation of epithelial mesenehymal tansition. Biochem Pharmcol, 60, 1091-9. https://doi.org/10.1016/S0006-2952(00)00427-5
  2. Desprez PY, Sumdia T, Coppe JP (2003). Helix-loop-helix proteins in mammary gland development and breast cancer. Mammary Gland Biol Neoplasia, 8, 225-39. https://doi.org/10.1023/A:1025957025773
  3. Haney SA (2007). Expanding the repertoire of RNA interference screens for developing new anticancer drug targets. Expert Opin Ther Targets, 11, 1429-41. https://doi.org/10.1517/14728222.11.11.1429
  4. Hasselblatt M, Mertsch S, Koos B, et al (2009). Twist-1 is overexpressed in neoplastic choroid plexus epithelial cells and promotes proliferation and invasion. Cancer Res, 69, 2219-23. https://doi.org/10.1158/0008-5472.CAN-08-3176
  5. Hay ED (2004). Tbe extracellular matrix in development and regeneration. Int J Dev Biol, 48, 687-94. https://doi.org/10.1387/ijdb.041857rt
  6. Jouppila-Mättö A, Närkiö-Mäkelä M, Soini Y, et al (2011). Twist and snai1 expression in pharyngeal squamous cell carcinoma stroma is related to cancer progression. BMC Cancer, 11, 350-63. https://doi.org/10.1186/1471-2407-11-350
  7. Kang Y, Massague J (2004). Epithelial mesenchymal transitions: twist in development and metastasis. Cell, 118, 277-9. https://doi.org/10.1016/j.cell.2004.07.011
  8. Kwok WK, Ling MT, Lee TW, et al (2005). Up-Regulation of Twist in Prostate Cancer and Its Implication as a Therapeutic Target. Cancer Res, 65, 5153-62. https://doi.org/10.1158/0008-5472.CAN-04-3785
  9. Lee JM, Dedhar S, Kalluri R, Thompson EW (2006). The epithelial-mesenchymal transition: new insights in signaling, development and disease. Cell Biol, 172, 973-81. https://doi.org/10.1083/jcb.200601018
  10. Matsuo N, Shiraha H, Fujikawa T, et al (2009). Twist expression promotes migration and invasion in hepatocellular carcinoma. BMC Cancer, 9, 240-55. https://doi.org/10.1186/1471-2407-9-240
  11. Niu RF, Zhang L, Xi GM, et al (2007). Up-regulation of Twist induces angiogenesis and correlates with metastasis in hepatocellular carcinoma. J Exp Clin Cancer Res, 26, 385-94.
  12. Puisieux A (2009). Role of epithelial-mesenchymal transition in tumor progressionm. Bull Acad Natl Med, 193, 2017-34.
  13. Radisky DC (2005). Epithelial-mesenchymal transition. Cell Sci, 118, 4325-6. https://doi.org/10.1242/jcs.02552
  14. Soo K, O'Rourke MP, Khoo PL, et al (2002). Twist function is required for the morphogenesis of the cephalic neural tube and the differentiation of the cranial neural crest cells in the mouse embryo. Dev Biol, 247, 251-70. https://doi.org/10.1006/dbio.2002.0699
  15. Thiery JP (2002). Epithelial-mesenchymal transitions in tumour progression. Nat Rev Cancer, 2, 442-54. https://doi.org/10.1038/nrc822
  16. Valdes-Mora F, Gomez del Pulgar T, Bandres E, et al (2009). Twist1 overexpression is associated with nodal invasion and male sex in primary colorectal cancer. Ann Surg Oncol, 16, 78-87. https://doi.org/10.1245/s10434-008-0166-x
  17. Valsesia-Wittmann S, Magdeleine M, Dupasquier S, et al (2004). Oncogenic cooperation between H-Twist and N-Myc overrides failsafe programs in cancer cells. Cancer Cell, 6, 625-30. https://doi.org/10.1016/j.ccr.2004.09.033
  18. Yang J, Mani SA, Dongaher JL, et al (2004). Twist, a master regulator of morphogenesis, plays an essential role in tumor metastasis. Cell, 117, 927-39. https://doi.org/10.1016/j.cell.2004.06.006
  19. Yoshida J, Hofiuchi A, Kikuchi N, et al (2009). Changes in the expression of E-cadherin repressers, Snail, Slug, SIP1, and Twist, in the development and progression of ovarian carcinoma: the important role of Snail in ovarian tumorigenesis and progresion. Med Mol Morphol, 42, 82-91. https://doi.org/10.1007/s00795-008-0436-5
  20. Yu AM (2007). Small interfering RNA in drug metabolism and transport. Curr Drug Metab, 8, 700-8. https://doi.org/10.2174/138920007782109751

Cited by

  1. Fisetin inhibits migration, invasion and epithelial-mesenchymal transition of LMP1-positive nasopharyngeal carcinoma cells vol.9, pp.2, 2013, https://doi.org/10.3892/mmr.2013.1836
  2. Role of epithelial to mesenchymal transition proteins in gynecological cancers: pathological and therapeutic perspectives vol.35, pp.10, 2014, https://doi.org/10.1007/s13277-014-2537-1
  3. Soluble factors regulated by epithelial-mesenchymal transition mediate tumour angiogenesis and myeloid cell recruitment vol.236, pp.4, 2015, https://doi.org/10.1002/path.4546
  4. Sonic hedgehog–Gli1 signals promote epithelial–mesenchymal transition in ovarian cancer by mediating PI3K/AKT pathway vol.32, pp.1, 2015, https://doi.org/10.1007/s12032-014-0368-y
  5. MiR-548c impairs migration and invasion of endometrial and ovarian cancer cells via downregulation of Twist vol.35, pp.1, 2016, https://doi.org/10.1186/s13046-016-0288-0
  6. Combined detection of Twist1, Snail1 and squamous cell carcinoma antigen for the prognostic evaluation of invasion and metastasis in cervical squamous cell carcinoma pp.1437-7772, 2017, https://doi.org/10.1007/s10147-017-1210-2
  7. Epithelial-to-Mesenchymal Transition and Cancer Invasiveness: What Can We Learn from Cholangiocarcinoma? vol.4, pp.12, 2015, https://doi.org/10.3390/jcm4121958
  8. A two-gene epigenetic signature for the prediction of response to neoadjuvant chemotherapy in triple-negative breast cancer patients vol.11, pp.1, 2019, https://doi.org/10.1186/s13148-019-0626-0