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Cucurbitacin-I, a Naturally Occurring Triterpenoid, Inhibits the CD44 Expression in Human Ovarian Cancer Cells

난소암 세포주의 CD44 발현에 미치는 Cucurbitacin-I의 효과

  • Seo, Hee Won (Department of Biomedical Science, Catholic University of Daegu) ;
  • Kim, Jin-Kyung (Department of Biomedical Science, Catholic University of Daegu)
  • 서희원 (대구가톨릭대학교 생명화학부 의생명과학전공) ;
  • 김진경 (대구가톨릭대학교 생명화학부 의생명과학전공)
  • Received : 2018.01.12
  • Accepted : 2018.06.04
  • Published : 2018.06.30

Abstract

Cucurbitacin-I, a natural triterpenoid derived from Cucurbitaceae family plants, exhibits a number of potentially useful pharmacological and biological activities. Indeed, the previous study demonstrated that cucurbitacin-I reduced the proliferation of colon cancer cells by enhancing apoptosis and causing cell cycle arrest at the G2/M phase. CD44, a type I transmembrane protein with the function of adhering to cells, mediates between the extracellular matrix and other cells through hyaluronic acid. Recent studies have demonstrated that an overexpression of the CD44 membrane receptor results in tumor initiation and growth, specific behaviors of cancer stem cells, the development of drug resistance, and metastasis. The aim was to examine the effect of cucurbitacin-I on CD44 expression human ovarian cancer cells because the effect of cucurbitacin-I on CD44 expression has not been reported. The expressions of CD44 mRNA and protein were detected using a quantitative real-time reverse-transcription polymerase chain reaction and a Western blot analysis, respectively. Treatment with cucurbitacin-I inhibited the expression of CD44 mRNA and protein. A subsequent analysis revealed that cucurbitacin-I blocked the phosphorylation of activator protein-1 (AP-1) and nuclear factor kappa-B ($NF-{\kappa}B$), which are key regulators of CD44 expression. Taken together, the data demonstrate that cucurbitacin-I regulates the AP-1 and $NF-{\kappa}B$ signaling pathways, leading to decreased CD44 expression.

박과 작물에 함유되어 있는 tetracyclic triterpene 성분 중 하나인 쿠쿠르비타신(cucurbitacin)-I는 대장암, 유방암, 간암세포에서의 항종양 활성이 밝혀져 있으나 난소암에서의 쿠쿠르비타신-I의 역할은 보고된 바 없다. CD44는 세포막에 존재하는 당단백질로 생체 내 리간드인 glycosaminoglycan hyaluronic acid를 통해 세포 외부 매트릭스와 다른 세포와의 접촉을 매개한다. 최근 연구에 의해 CD44의 발현이 난소암세포의 증식 및 세포 부착과 침윤을 증가시키는 주요 원인이라는 것이 보고되었다. 이러한 결과는 CD44의 발현을 억제함으로써 난소암의 진행을 조절할 수 있음을 시사하고 있다. 본 연구에서는 쿠쿠르비타신-I가 난소암세포의 CD44의 발현을 억제할 수 있는 지의 여부를 조사하였다. 인간의 난소암 세포인 SKOV-3를 이용한 MTS assay를 수행한 결과, 쿠쿠르비타신-I는 100 nM이상의 농도에서 세포독성을 나타내었다. 세포독성을 나타내지 않는 농도의 쿠쿠르비타신-I를 SKOV-3 세포에 처리하여 Western blot 분석과 qRT-PCR을 수행한 결과, 쿠쿠르비타신-I에 의해 CD44의 단백질과 mRNA의 발현이 유의적으로 감소되는 것을 확인하였다. 또한 쿠쿠르비타신-I에 의한 CD44의 발현 억제가 $NF-{\kappa}B$와 AP-1의 인산화 감소에 기인하고 있음을 밝혔다. 이러한 결과는 쿠쿠르비타신-I가 CD44 발현을 억제하는 기능을 가지며, 이는 난소암 치료에 도움을 줄 수 있는 제재로서 쿠쿠르비타신-I의 가능성을 제시하는 것이다.

Keywords

References

  1. Aruffo, A., Stamenkovic, I., Melnick, M., Underhill, C. B. and Seed, B. 1990. CD44 is the principal cell surface receptor for hyaluronate. Cell 61, 1303-1313. https://doi.org/10.1016/0092-8674(90)90694-A
  2. Bajorath, J., Greenfield, B., Munro, S. B., Day, A. J. and Aruffo, A. 1998. Identification of CD44 residues important for hyaluronan binding and delineation of the binding site. J. Biol. Chem. 273, 338-343. https://doi.org/10.1074/jbc.273.1.338
  3. Blaskovich, M. A., Sun, J., Cantor, A., Turkson, J., Jove, R. and Sebti, S. M. 2003. Discovery of JSI-124 (cucurbitacin I), a selective Janus kinase/signal transducer and activator of transcription 3 signaling pathway inhibitor with potent antitumor activity against human and murine cancer cells in mice. Cancer Res. 63, 1270-1279.
  4. Bourguignon, L. Y., Peyrollier, K., Xia, W. and Gilad, E. 2008. Hyaluronan-CD44 interaction activates stem cell marker Nanog, Stat-3-mediated MDR1 gene expression, and ankyrin-regulated multidrug efflux in breast and ovarian tumor cells. J. Biol. Chem. 283, 17635-17651. https://doi.org/10.1074/jbc.M800109200
  5. Chen, Y. W., Chen. K. H., Huang, P. I., Chen, Y. C., Chiou, G. Y., Lo, W. L., Tseng, L. M., Hsu, H. S., Chang, K. W. and Chiou, S. H. 2010. Cucurbitacin I suppressed stem-like property and enhanced radiation-induced apoptosis in head and neck squamous carcinoma-derived $CD44^+\;ALDH1^+$cells. Mol. Cancer Ther. 9, 2879-2892. https://doi.org/10.1158/1535-7163.MCT-10-0504
  6. Cortez, A. J., Tudrej, P., Kujawa, K. A. and Lisowska, K. M. 2018. Advances in ovarian cancer therapy. Cancer Chemother. Pharmacol. 81, 17-38. https://doi.org/10.1007/s00280-017-3501-8
  7. Gao, Y., Foster, R., Yang, X., Feng, Y., Shen, J. K., Mankin, H. J., Hornicek, F. J., Amiji, M. M. and Duan, Z. 2015. Up-regulation of CD44 in the development of metastasis, recurrence and drug resistance of ovarian cancer. Oncotarget 6, 9313-9326.
  8. Jia, Q., Feng, M., Wang, Y. and Xue, S. 2008. Gastric cancer cells in collagen gel matrix: Three-dimensional growth and differential expression of adhesion molecules (CD44s, CD54, E-cadherin). J. Biomed. Mater. Res. A. 84, 917-925.
  9. Kim, H. J., Park, J. H. and Kim, J. K. 2014. Cucurbitacin-I, a natural cell-permeable triterpenoid isolated from Cucurbitaceae, exerts potent anticancer effect in colon cancer. Chem. Biol. Interact. 219, 1-8. https://doi.org/10.1016/j.cbi.2014.05.005
  10. Lui, V. W., Yau, D. M., Wong, E. Y., Ng ,Y. K., Lau, C. P., Ho, Y., Chan, J. P., Hong, B., Ho, K., Cheung, C. S., Tsang, C. M., Tsao, S. W. and Chan, A. T. 2009. Cucurbitacin I elicits anoikis sensitization, inhibits cellular invasion and in vivo tumor formation ability of nasopharyngeal carcinoma cells. Carcinogenesis 30, 2085-2094. https://doi.org/10.1093/carcin/bgp253
  11. Mao, M., Zheng, X., Jin, B., Zhang, F., Zhu, L. and Cui, L. 2017. Effects of CD44 and E-cadherin overexpression on the proliferation, adhesion and invasion of ovarian cancer cells. Exp. Ther. Med. 14, 5557-5563.
  12. Ren, Y., Yu, K., Sun, S., Li, Z., Yuan, J., Han, X. D., Shi, J. and Zhen, L. 2014. JSI124 inhibits breast cancer cell growth by suppressing the function of B cells via the downregulation of signal transducer and activator of transcription 3. Oncol. Lett. 8, 928-932. https://doi.org/10.3892/ol.2014.2221
  13. Sacks, J. D. and Barbolina, M. V. 2015. Expression and function of CD44 in epithelial ovarian carcinoma. Biomolecules 5, 3051-3066. https://doi.org/10.3390/biom5043051
  14. Shi, Y. Y. and Jiang, H. 2016. Prognostic role of the cancer stem cell marker CD44 in ovarian cancer: A meta-analysis. Genet. Mol. Res. 15, doi: 10.4238/gmr.15038325.
  15. Smith, S. M., Lyu, Y. L. and Cai, L. 2014. NF-${\kappa}B$ affects proliferation and invasiveness of breast cancer cells by regulating CD44 expression. PLoS One 9, e106966. https://doi.org/10.1371/journal.pone.0106966
  16. Song, J., Liu, H., Li, Z., Yang, C. and Wang, C. 2015. Cucurbitacin I inhibits cell migration and invasion and enhances chemosensitivity in colon cancer. Oncol. Rep. 33, 1867-1871. https://doi.org/10.3892/or.2015.3749
  17. Su, Y., Li, G., Zhang, X., Gu, J., Zhang, C., Tian, Z. and Zhang, J. 2008. JSI-124 inhibits glioblastoma multiforme cell proliferation through G(2)/M cell cycle arrest and apoptosis augment. Cancer Biol. Ther. 7, 1243-1249. https://doi.org/10.4161/cbt.7.8.6263