Apoptosis induction of human endometriotic epithelial and stromal cells by noscapine

Document Type: Original Article


1 Fertility and Infertility Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran

2 Department of Anatomy, Medical Faculty, Tehran University of Medical Sciences, Tehran, Iran


Objective(s): Endometriosis is a complex gynecologic disease with unknown etiology. Noscapine has been introduced as a cancer cell suppressor. Endometriosis was considered as a cancer like disorder, The aim of present study was to investigate noscapine apoptotic effect on human endometriotic epithelial and stromal cells in vitro.
Materials and Methods:In this in vitro study, endometrial biopsies from endometriosis patients (n=9) were prepared and digested by an enzymatic method (collagenase I, 2 mg/ml). Stromal and epithelial cells were separated by sequential filtration through a cell strainer and ficoll layering. The cells of each sample were divided into five groups: control (0), 10, 25, 50 and 100 micromole/liter (µM) concentration of noscapine and were cultured for three different periods of times; 24, 48 and 72 hr. Cell viability was assessed by colorimetric assay. Nitric oxide (NO) concentration was measured by Griess reagent. Cell death was analyzed by Acridine Orange (AO)–Ethidium Bromide (EB) double staining and Terminal deoxynucleotidyl transferase (TdT) dUTP Nick-End Labeling (TUNEL) assay. Data were analyzed by one-way ANOVA.
Results: Viability of endometrial epithelial and stromal cells significantly decreased in 10, 25, 50 and 100 µM noscapine concentration in 24, 48, 72 hr (P<0.05) and apoptotic index increased in 25, 50 and 100 µM noscapine concentrations in 48 hr significantly (P<0.05). Concentrations of NO didn’t show a significant decrease.
Conclusion: Noscapine increased endometriotic epithelial and stromal cell death and can be suggested as a treatment for endometriosis.


1. Dunselman GA, Vermeulen N, Becker C, Calhaz-Jorge C, D'Hooghe T, De Bie B, et al ESHRE guideline: management of women with endometriosis. European Society of Human Reproduction and Embryology. Hum Reprod 2014; 29:400-412.

2.  Vignali M, Infantino M, Matronem R, Chiodo I, Somigliana E, Busacca M, et al Endometriosis: novel etiopathogenetic concepts and clinical perspectives. Fertil Steril 2002; 78; 665–678.

3. Nap AW, Groothuis PG, Demir AY, Evers JL, Dunselman GA. Pathogenesis of endometriosis. Best Practice and Research Clinical Obstetrics and Gynaecolgy 2004; 18: 233–244.

4. Giudice LC, Kao LC.  Endometriosis. Lancet 2004; 364:1789-1799.

5. Yadav L, Puri N, Rastogi V, Satpute P, Sharma V. Tumour Angiogenesis and Angiogenic Inhibitors: A Review. J Clin Diagn Res. 2015; 9: XE01–XE05.

6. Taylor RN, Lebovic DI. Mueller MD. Angiogenic factors in endometriosis. Annals of New York Academy Sciences 2002; 955; 89-100.

7. Sainz de la Cuesta R, Eichhorn JH, Rice LW, Fuller AF, Nikrui N, Goff BA. Histologic transformation of benign endometriosis to early epithelial ovarian cancer. Gynecol Oncol. 1996; 60: 238–244.

8. Huang HY. Medical Treatment of Endometriosis. Chang Gung Med J. 2008; 31: 431-440.

9. Vercellini P, De Giorgi O, Mosconi P, Stellato, G, Vicentini S, Crosignani PG. Cyproterone acetate versus a continuous monophasic oral contraceptive in the treatment of recurrent pelvic pain after conservative surgery for symptomatic endometriosis. Fertil Steril 2002; 77: 52- 61.

10. Kettel LM, Murphy AA, Morales AJ, Yen SS. Preliminary report on the treatment of endometriosis with low-dos mifepristone (RU 486). Am J Obstet Gynecol 1998; 178: 1151-1156.

11. Esfandiari N, Khazaei M, Jafar A, Bielecki R, Gotlieb L, Ryan E. et al Effect of a statin on an in vitro model of endometriosis. Fertil Steril 2007; 87: 257–262.

12. Rezavand N, Khazaei M, Oliapanah E, Nikzad H, Khazaei MR. Low doses of celecoxib stimulate human endometrium growth in a three-dimensional culture model. Int J Fertil Steril 2013; 7: 7-12.

13. Kokawa K, Shikone T, Nakano R. Apoptosis in the human uterine endometrium during the menstrual cycle. J Clinic Endocrin Met 1996; 81: 4144-4147.

14.  Meresman GF, Bilotas MA, Lombardi E, Tesone M, Sueldo C, Baranao R. Effect of GnRH analogues on apoptosis and release of interleukin-1b and vascular endothelial growth factor in endometrial cell cultures from patients with endometriosis. Hum Reprod 2003; 18: 1767-1771.

15. Mahmoudian M. Recent progress in clinical application of noscapine: a review. Cur Topics Pharmacology 2006; 10: 81-86.

16. Ebrahimi SA, Zareie-Rostami P, Mahmoudian M. Interaction of noscapine with the bradukinin mediation of cough response. Acta Physiolo 2003; 90: 147-155.

17- Landen   JW, Lang   R, McMahon   SJ,  Rusan NM, Yvon AM, Adams AW, et al Noscapine alters microtubule dynamics in living cells and inhibits the progression of melanoma. Cancer Research 2002; 62: 4109-4114.

18. Aneja R, Ghaleb AM, Zhou J, Yang VW.  Joshi HC. P53 and p21 determine the sensitivity of noscapine-induced apoptosis in colon cancer cells.  Cancer Research 2007; 67: 3862-3870.

19. Ye K, Ke Y, Keshava N, Shanks J, Kapp JA, Tekmal RR, et al Opium alkaloid noscapine is an antitumor agent that arrest metaphase and induces apoptosis in dividing cells. Proceeding of the National Academy of Sciences USA 1998; 95: 1601-1606.

20- Mahmoudian M. Rahimi-Moghaddam P. The anti-cancer activity of noscapine: a review.  Recent Patents on Anticancer Drug Discovery 2009; 4: 92-97.

21. Barken I, Geller J, Rogosnitzky M.  Noscapine inhibits human prostate cancer progression and metastasis in a mouse model.  Anticancer Research 2008; 28: 3701-3704.

22. Khazaei M, Chobsaz F, Khazaei S. The effect of different doses of clomiphene citrate on morphology and proliferation of endometrial stromal cells in in-vitro culture. J Bab Uni Med Sci. 2010; 12: 7-12.

23. Tayarani-Najaran Z, Parsaee H,  Hoseini A. Mousavi SH. Study of noscapine-induced cell death in hepatocellular carcinoma cell Line. Pharmacology online 2009; 3: 522-530.

24. Khazaei M, Roshankhah Sh, Ghorbani P.  Chobsaz F. Sildenafil effect on nitric oxide secretion by normal human endometrial epithelial cells cultured in vitro. Int J Fertil Steril 2011; 5; 142-147.

25. Mirzapur P, Rashidi Z, Rezakhani L, Khazaei M.  In vitro inhibitory effect of crab shell extract on human umbilical vein endothelial cell. In Vitro Cellular & Developmental Biology –Animal 2015; 51: 36-41.

26. Baskic D, Popovic S, Ristic P, Nebojsa N. Analysis of cycloheximide-induced apoptosis in human leukocytes: Fluorescence microscopy using annexin V/propidium iodide versus acridin orange/ethidium bromide.  Cell Biol Intern 2006; 30, 924-932.

27. Rashidi Z, Azadbakht M, Khazaei M. Hydrostatic pressure improves in-vitro maturation of oocytes derived from vitrified-warmed mouse ovaries. Iran J Reprod Med. 2012; 10: 257-64.

28. Chougule MB, Apurva RP, Jackson T, Patel R. Antitumor activity of noscapine in combination with doxorubicin in triple negative breast cancer. Mandip Singh PLoS ONE  2011; 3: 1-12.   

29. Ronald l Steve JM, Jaren W, McMahon J. Noscapine alters microtubule dynamics and inhibits the progression of melanoma in living cells. Cancer Res 2002; 62: 4109-4114.

30. Zhou J, Gupta K, Aggarwal S, Aneja R, Chandra R, Panda D, et al Brominated derivatives of noscapine are potent microtubule-interfering agents that perturb mitosis and inhibit cell proliferation.  Molecular Pharmacology 2003; 63: 799-807.

31. Heidari N, Goliaei B, Moghaddam PR, Rahbar-Roshandel N, Mahmoudian M. Apoptotic pathway induced by noscapine in human myelogenous leukemic cells.  Anticancer Drugs 2007; 18: 1139-1147.

32. Wu MY, Chao KH, Yang JH, Lee TH, Yang YS,  Ho HN, Nitric oxide synthesis is increased in the endometrial tissue of women with endometriosis. Hum Reprod 2003; 18: 2668- 2671. 

33. BakanE, Taysi S, Polat MF, Dalga Z, Bakan N, Gumus M. Nitric oxide levels and lipid peroxidation in plasma of patients with gastric cancer. Clinical Oncology 2002; 32:162-166.

34. Tong X. Li H.  eNOS protects prostate cancer cells from TRAIL-induced apoptosis. Cancer Letters 2004; 210: 63–71. 

35. Vaskivuo TE, Stenbäck F, Karhumaa P, Risteli J, Dunkel L. Tapanainen JS. Apoptosis and apoptosis-related proteins in human endometrium. Molecular Cellular Endocrinology 2000; 165: 75-83.

36. Dahmoun M,  Boman K, Cajander S, Westin P,  Bäckström T. Apoptosis, proliferation and sex hormone receptors in superficial parts of human endometrium at the end of the secretory phase.  J  Clin Endocrinol Meta 1999; 84: 1737-1743.

37. Osborn BH, Haney AF, Misukonis MA, Weinberg JB. Inducible nitric oxide synthase expression by peritoneal macrophages in endometriosis-associated infertility. Fertil Steril 2002; 77: 46-51.

38. Gebel  HM, Braun DP, Tambur A, Frame D, Rana N, Dmowski WP.  Spontaneous apoptosis of endometrial tissue is impaired in women with endometriosis. Fertil Steril  1998; 69: 1042-1047.

39. Dmowski WP, Ding J, Shen J, Rana N, Fernandez BB, Braun DP. Apoptosis in endometrial glandular and stromal cells in women with and without endometriosis. Hum Reprod 2001; 16: 1802-1808.

40. Dunselman GA, Hendrix MG, Bouckaert PX, Evers JL. Functional aspects of peritoneal macrophages in endometriosis of women. J Reprod Fertil 1988; 82: 707-710.

41. Fasciani A1, Bocci G, Xu J, Bielecki R, Greenblatt E, Leyland N, et al Three-dimensional in vitro culture of endometrial explants mimics the early stages of endometriosis. Fertil Steril. 2003; 80:1137-1143.