DOI QR코드

DOI QR Code

The Inhibition Effect of Triptolide on Human Endometrial Carcinoma Cell Line HEC-1B: a in vitro and in vivo Studies

  • Ni, Jing (Department of Gynecologic Oncology, the Affiliated Cancer Hospital of Nanjing Medical University and Jiangsu Cancer Hospital) ;
  • Wu, Qiang (Department of Gynecologic Oncology, the Affiliated Cancer Hospital of Nanjing Medical University and Jiangsu Cancer Hospital) ;
  • Sun, Zhi-Hua (Department of Radiation of Gynecologic Oncology, the Affiliated Cancer Hospital of Nanjing Medical University and Jiangsu Cancer Hospital) ;
  • Zhong, Jian (Department of Gynecologic Oncology, the Affiliated Cancer Hospital of Nanjing Medical University and Jiangsu Cancer Hospital) ;
  • Cai, Yu (Department of Gynecologic Oncology, the Affiliated Cancer Hospital of Nanjing Medical University and Jiangsu Cancer Hospital) ;
  • Huang, Xin-En (Department of Chemotherapy, the Affiliated Cancer Hospital of Nanjing Medical University and Jiangsu Cancer Hospital)
  • Published : 2015.06.26

Abstract

Background: To investigate the inhibitory effect and the underlying mechanism of triptolide on cultured human endometrial carcinoma HEC-1B cells and corresponding xenograft. Materials and Methods: For in vitro studies, the inhibition effect of proliferation on HEC-1B cell by triptolide was determined by MTT assay; cell cycle and apoptosis of the triptolide-treated and untreated cells were detected by flow cytometry. For in vivo studies, a xenograft tumor model of human endometrial carcinoma was established using HEC-1B cells, then the tumor-bearing mice were treated with high, medium, and low-dose ($8{\mu}g$, $4{\mu}g$ and $2{\mu}g/day$) triptolide or cisplatin at $40{\mu}g/day$ or normal saline as control. The mice were treated for 10-15 days, during which body weight of the mice and volume of the xenograft were weighted. Then expression of Bcl-2 and vascular endothelial growth factor (VEGF) was analyzed by SABC immunohistochemistry. Results: Cell growth was significantly inhibited by triptolide as observed by an inverted phase contrast microscope; the results of MTT assay indicated that triptolide inhibits HEC-1B cell proliferation in a dose and time-dependent manner; flow cytometry showed that low concentration (5 ng/ml) of triptolide induces cell cycle arrest of HEC-1B cells mainly at S phase, while higher concentration (40 or 80 ng/ml) induced cell cycle arrest of HEC-1B cells mainly at G2/M phase, and apoptosis of the cells was also induced. High-dose triptolide showed a similar tumor-inhibitory effect as cisplatin (-50%); high-dose triptolide significantly inhibited Bcl-2 and VEGF expression in the xenograft model compared to normal saline control (P<0.05). Conclusions: triptolide inhibits HEC-1B cell growth both in vitro and in mouse xenograft model. Cell cycle of the tumor cells was arrested at S and G2/M phase, and the mechanism may involve induction of tumor cell apoptosis and inhibition of tumor angiogenesis.

Keywords

Uterine cancer;triptolide;VEGF;Bcl-2

Acknowledgement

Supported by : Research Foundation of Jiangsu Cancer Hospital

References

  1. Zhong YY, Chen HP, Tan BZ, Yu HH, Huang XS (2013). Triptolide avoids cisplatin resistance and induces apoptosis via the reactive oxygen species/nuclear factor-${\kappa}B$ pathway in SKOV3PT platinum-resistant human ovarian cancer cells. Oncol Lett, 6, 1084-92. https://doi.org/10.3892/ol.2013.1524
  2. Alsaied OA, Sangwan V, Banerjee S, et al (2014). Sorafenib and triptolide as combination therapy for hepatocellular carcinoma. Surgery, 156, 270-9. https://doi.org/10.1016/j.surg.2014.04.055
  3. Cai YY, Lin WP, Li AP, Xu JY (2013). Combined effects of curcumin and triptolide on an ovarian cancer cell line. Asian Pac J Cancer Prev, 14, 4267-71. https://doi.org/10.7314/APJCP.2013.14.7.4267
  4. Chen RH, Tian YJ (2013). Enhanced anti-tumor efficacy of aspirin combined with triptolide in cervical cancer cells. Asian Pac J Cancer Prev, 14, 3041-4. https://doi.org/10.7314/APJCP.2013.14.5.3041
  5. Dobrzycka B, Mackowiak-Matejczyk B, Kinalski M, Terlikowski SJ (2013). Pretreatment serum levels of bFGF and VEGF and its clinical significance in endometrial carcinoma. Gynecol Oncol, 128, 454-60. https://doi.org/10.1016/j.ygyno.2012.11.035
  6. Ho JN, Byun SS, Lee S, et al (2015). Synergistic antitumor effect of triptolide and cisplatin in cisplatin resistant human bladder cancer cells. J Urol, 193, 1016-22. https://doi.org/10.1016/j.juro.2014.09.007
  7. Kwon HY, Kim KS, An HK, et al (2013). Triptolide induces apoptosis through extrinsic and intrinsic pathways in human osteosarcoma U2OS cells. Indian J Biochem Biophys, 50, 485-91.
  8. Li CJ, Chu CY, Huang LH, et al (2012). Synergistic anticancer activity of triptolide combined with cisplatin enhances apoptosis in gastric cancer in vitro and in vivo. Cancer Lett. 319, 203-13. https://doi.org/10.1016/j.canlet.2012.01.006
  9. Li J, Liu R, Yang Y, et al (2014). Triptolide-induced in vitro and in vivo cytotoxicity in human breast cancer stem cells and primary breast cancer cells. Oncol Rep, 31, 2181-6. https://doi.org/10.3892/or.2014.3115
  10. Liu Y, Xiao E, Yuan L, Li G (2014). Triptolide synergistically enhances antitumor activity of oxaliplatin in colon carcinoma in vitro and in vivo. DNA Cell Biol, 33, 418-25. https://doi.org/10.1089/dna.2014.2356
  11. Park B (2014). Triptolide, a diterpene, inhibits osteoclastogenesis, induced by RANKL signaling and human cancer cells. Biochimie, 105, 129-36. https://doi.org/10.1016/j.biochi.2014.07.003
  12. Saarelainen SK, Staff S, Peltonen N, et al (2014). Endoglin, VEGF, and its receptors in predicting metastases in endometrial carcinoma. Tumour Biol, 35, 4651-7. https://doi.org/10.1007/s13277-014-1609-6
  13. Siegel R1, Ma J, Zou Z, Jemal A (2014). Cancer statistics, 2014. CA Cancer J Clin, 64, 9-29. https://doi.org/10.3322/caac.21208
  14. Wang J, Taylor A, Showeil R, et al (2014). Expression profiling and significance of VEGF-A, VEGFR2, VEGFR3 and related proteins in endometrial carcinoma. Cytokine. 68, 94-100. https://doi.org/10.1016/j.cyto.2014.04.005
  15. Wang W, Lin W, Hong B, et al (2012). Effect of triptolide on malignant peripheral nerve sheath tumours in vitro and in vivo. J Int Med Res, 40, 2284-94. https://doi.org/10.1177/030006051204000626
  16. Zhao H, Yang Z, Wang X, et al (2012). Triptolide inhibits ovarian cancer cell invasion by repression of matrix metalloproteinase 7 and 19 and upregulation of E-cadherin. Exp Mol Med, 44, 633-41. https://doi.org/10.3858/emm.2012.44.11.072

Cited by

  1. A guide for endometrial cancer cell lines functional assays using the measurements of electronic impedance pp.1573-0778, 2017, https://doi.org/10.1007/s10616-017-0149-5
  2. Triptolide Combined with Radiotherapy for the Treatment of Nasopharyngeal Carcinoma via NF-κB-Related Mechanism vol.17, pp.12, 2016, https://doi.org/10.3390/ijms17122139