Analysis of the in vitro effects of di‑(2‑ethylhexyl) phthalate exposure on human uterine leiomyoma cells

Kim,Jin  Hee

doi:10.3892/etm.2018.6040

Analysis of the in vitro effects of di‑(2‑ethylhexyl) phthalate exposure on human uterine leiomyoma cells

Authors:
- Jin Hee Kim
View Affiliations

Affiliations: Department of Internal Medicine, Seoul National University Bundang Hospital, Seongnam, Gyeonggi 13620, Republic of Korea
Published online on: April 10, 2018 https://doi.org/10.3892/etm.2018.6040
Pages: 4972-4978

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Uterine leiomyoma is the most common benign tumor type of the female reproductive tract. Despite its high prevalence, the exact pathogenesis of the benign tumor remains unknown. In the present study, the effects of di‑(2‑ethylhexyl) phthalate (DEHP) on the proliferation and apoptosis rates and expression of inflammatory proteins in human leiomyoma cells were evaluated. The effects of DEHP on cell viability were determined by 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium bromide assay. The effects on apoptosis were evaluated by western blotting, TUNEL assay and Annexin V staining. Western blotting was also performed to evaluate the expression of inflammatory proteins. It was observed that DEHP‑treated leiomyoma cells had higher viability, as well as proliferating cell nuclear antigen and B‑cell lymphoma 2 protein expression, and lower apoptosis rates compared with the untreated controls. Additionally, hypoxia inducible factor 1α (HIF‑1α) and cyclooxygenase‑2 (COX‑2) expression increased in human leiomyoma cells following DEHP treatment. In conclusion, DEHP promoted cell viability and anti‑apoptotic protein expression and induced HIF‑1α and COX‑2 expression in human leiomyoma cells. These results suggested that DEHP may disrupt mechanisms underlying various processes in human leiomyoma cells. Furthermore, the current study revealed a basic mechanism of action of DEHP in human leiomyoma cells. Further research on the effects of various endocrine disruptors on the pathogenesis of uterine leiomyoma during early development may reveal strategies to prevent this disease.

View Figures

View References

1	Falcone T and Walters MD: Hysterectomy for benign disease. Obstet Gynecol. 111:753–767. 2008. View Article : Google Scholar : PubMed/NCBI
2	Parker WH: Etiology, symptomatology, and diagnosis of uterine myomas. Fertil Steril. 87:725–736. 2007. View Article : Google Scholar : PubMed/NCBI
3	Sell SM, Tullis C, Stracner D, Song CY and Gewin J: Minimal interval defined on 7q in uterine leiomyoma. Cancer Genet Cytogenet. 157:67–69. 2005. View Article : Google Scholar : PubMed/NCBI
4	Rose DP, Goldman M, Connolly JM and Strong LE: High-fiber diet reduces serum estrogen concentrations in premenopausal women. Am J Clin Nutr. 54:520–525. 1991. View Article : Google Scholar : PubMed/NCBI
5	Prentice R, Thompson D, Clifford C, Gorbach S, Goldin B and Byar D: Dietary fat reduction and plasma estradiol concentration in healthy postmenopausal women. The women's health trial study group. J Natl Cancer Inst. 82:129–134. 1990. View Article : Google Scholar : PubMed/NCBI
6	Greenberg MD and Kazamel TI: Medical and socioeconomic impact of uterine fibroids. Obstet Gynecol Clin North Am. 22:625–636. 1995.PubMed/NCBI
7	Wang F, Chen J, Wang L, Ma Y and Mayinuer N: CYP1A1 genetic polymorphisms and uterine leiomyoma risk: A meta-analysis. Int J Clin Exp Med. 8:3590–3594. 2015.PubMed/NCBI
8	Markey CM, Rubin BS, Soto AM and Sonnenschein C: Endocrine disruptors: From Wingspread to environmental developmental biology. J Steroid Biochem Mol Biol. 83:235–244. 2002. View Article : Google Scholar : PubMed/NCBI
9	Yu L, Moore AB and Dixon D: Receptor tyrosine Kinases and their hormonal regulation in uterine leiomyoma. Semin Reprod Med. 28:250–259. 2010. View Article : Google Scholar : PubMed/NCBI
10	Sharpe RM, Fisher JS, Millar MM, Jobling S and Sumpter JP: Gestational and lactational exposure of rats to xenoestrogens results in reduced testicular size and sperm production. Environ Health Perspect. 103:1136–1143. 1995. View Article : Google Scholar : PubMed/NCBI
11	Lin Y, Wei J, Li Y, Chen J, Zhou Z, Song L, Wei Z, Lv Z, Chen X, Xia W and Xu S: Developmental exposure to di(2-ethylhexyl) phthalate impairs endocrine pancreas and leads to long-term adverse effects on glucose homeostasis in the rat. Am J Physiol Endocrinol Metab. 301:E527–E538. 2011. View Article : Google Scholar : PubMed/NCBI
12	Tomaszewski KE, Heindel SW, Jenkins WL and Melnick RL: Induction of peroxisomal acyl CoA oxidase activity and lipid peroxidation in primary rat hepatocyte cultures. Toxicology. 17:49–60. 1990. View Article : Google Scholar
13	Halden RU: Plastics and health risks. Annu Rev Public Health. 31:179–194. 2010. View Article : Google Scholar : PubMed/NCBI
14	Ha M, Guan X, Wei L, Li P, Yang M and Liu C: Di-(2-ethylhexyl) phthalate inhibits testosterone level through disturbed hypothalamic-pituitary-testis axis and ERK-mediated 5α-Reductase 2. Sci Total Environ. 563–564:566–575. 2016. View Article : Google Scholar
15	Tseng IL, Yang YF, Yu CW, Li WH and Liao VH: Phthalates induce neurotoxicity affecting locomotor and thermotactic behaviors and AFD neurons through oxidative stress in Caenorhabditis elegans. PLoS One. 8:e826572013. View Article : Google Scholar : PubMed/NCBI
16	Caldwell JC: DEHP: Genotoxicity and potential carcinogenic mechanisms-a review. Mutat Res. 751:82–157. 2012. View Article : Google Scholar : PubMed/NCBI
17	McKee RH, Butala JH, David RM and Gans G: NTP center for the evaluation of risks to human reproduction reports on phthalates: Addressing the data gaps. Reprod Toxicol. 18:1–22. 2004. View Article : Google Scholar : PubMed/NCBI
18	Hoyer PB: Reproductive toxicology: Current and future directions. Biochem Pharmacol. 62:1557–1564. 2001. View Article : Google Scholar : PubMed/NCBI
19	Wang GL, Jiang BH, Rue EA and Semenza GL: Hypoxia-inducible factor1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension. Proc Natl Acad Sci USA. 92:5510–5514. 1995. View Article : Google Scholar : PubMed/NCBI
20	Maxwell PH, Wiesener MS, Chang GW, Clifford SC, Vaux EC, Cockman ME, Wykoff CC, Pugh CW, Maher ER and Ratcliffe PJ: The tumour suppressor protein VHL targets hypoxia-inducible factors for oxygen-dependent proteolysis. Nature. 399:271–275. 1999. View Article : Google Scholar : PubMed/NCBI
21	Harris AL: Hypoxia-a key regulatory factor in tumour growth. Nat Rev Cancer. 2:38–47. 2002. View Article : Google Scholar : PubMed/NCBI
22	Csiki I, Yanagisawa K, Haruki N, Nadaf S, Morrow JD, Johnson DH and Carbone DP: Thioredoxin-1 modulates transcription of cyclooxygenase-2 via hypoxia-inducible factor-1alpha in non-small cell lung cancer. Cancer Res. 66:143–150. 2006. View Article : Google Scholar : PubMed/NCBI
23	Cao Y and Prescott SM: Many actions of cyclooxygenase-2 in cellular dynamics and in cancer. J Cell Physiol. 190:279–286. 2002. View Article : Google Scholar : PubMed/NCBI
24	Williams CS, Mann M and DuBois RN: The role of cyclooxygenases in inflammation, cancer, and development. Oncogene. 18:7908–7916. 1999. View Article : Google Scholar : PubMed/NCBI
25	Williams C, Shattuck-Brandt RL and DuBois RN: The role of COX-2 in intestinal cancer. Ann N Y Acad Sci. 889:72–83. 1999. View Article : Google Scholar : PubMed/NCBI
26	Dempke W, Rie C, Grothey A and Schmoll HJ: Cyclooxygenase-2: A novel target for cancer chemotherapy? J Cancer Res Clin Oncol. 127:411–417. 2001. View Article : Google Scholar : PubMed/NCBI
27	Singer CA, Baker KJ, McCaffrey A, AuCoin DP, Dechert MA and Gerthoffer WT: p38MAPK and NF-kappaB mediate COX-2 expression in human airway myocytes. Am J Physiol Lung Cell Mol Physiol. 2855:L1087–L1098. 2003. View Article : Google Scholar
28	Swartz CD, Afshari CA, Yu L, Hall KE and Dixon D: Estrogen-induced changes in IGF-I, Myb family and MAP kinase pathway genes in human uterine leiomyoma and normal uterine smooth muscle cell lines. Mol Hum Reprod. 11:441–450. 2005. View Article : Google Scholar : PubMed/NCBI
29	Colón I, Caro D, Bourdony CJ and Rosario O: Identification of phthalate esters in the serum of young Puerto Rican girls with premature breast development. Environ Health Perspect. 108:895–900. 2000. View Article : Google Scholar
30	Durmaz E, Ozmert EN, Erkekoglu P, Giray B, Derman O, Hincal F and Yurdakök K: Plasma phthalate levels in pubertal gynecomastia. Pediatrics. 125:e122–e129. 2010. View Article : Google Scholar : PubMed/NCBI
31	Wittassek M and Angerer J: Phthalates: Metabolism and exposure. Int J Androl. 31:131–138. 2008. View Article : Google Scholar : PubMed/NCBI
32	Chen FP and Chien MH: Lower concentrations of phthalates induce proliferation inhuman breast cancer cells. Climacteric. 17:377–384. 2014. View Article : Google Scholar : PubMed/NCBI
33	Martinasso G, Maggiora M, Trombetta A, Canuto RA and Muzio G: Effects of di(2-ethylhexyl)phthalate, a widely used peroxisome proliferator and plasticizer, oncell growth in the human keratinocyte cell line NCTC 2544. J Toxic Environ Health A. 69:353–365. 2006. View Article : Google Scholar
34	Chen X, Qin Q, Zhang W, Zhang Y, Zheng H, Liu C, Yang Y, Xiong W and Yuan J: Activation of thepi3k-akt-mtor signaling pathway promotes dehp-induced hep3b cell proliferation. Food Chem Toxicol. 59:325–333. 2013. View Article : Google Scholar : PubMed/NCBI
35	Zhu H, Zheng J, Xiao X, Zheng S, Dong K, Liu J and Wang Y: Environmental endocrine disruptors promote invasion and metastasis of SK-N-SH human neuroblastoma cells. Oncol Rep. 23:129–139. 2010.PubMed/NCBI
36	Jepsen KF, Abildtrup A and Larsen ST: Monophthalates promote IL-6 and IL-8production in the human epithelial cell line A549. Toxicol In Vitro. 18:265–269. 2004. View Article : Google Scholar : PubMed/NCBI
37	Burmeister A, Assi LK, Ferro CJ, Hughes RG, Barnett AH, Bellary S, Cockwell P, Pratt G and Hutchison CA: The relationship between high-sensitivity CRP and polyclonal free light chains as markers of inflammation in chronic disease. Int J Lab Hematol. 36:415–424. 2014. View Article : Google Scholar : PubMed/NCBI
38	Ferguson KK, Loch-Caruso R and Meeker JD: Urinary phthalatemetabolites inrelation to biomarkers of inflammation and oxidative stress: NHANES 1999–2006. Environ Res. 111:718–726. 2011. View Article : Google Scholar : PubMed/NCBI
39	Huang PC, Tsai EM, Li WF, Liao PC, Chung MC, Wang YH and Wang SL: Associationbetween phthalate exposure and glutathione S-transferase M1 polymorphism in adenomyosis, leiomyoma and endometriosis. Hum Reprod. 25:986–994. 2010. View Article : Google Scholar : PubMed/NCBI
40	Palomba S, Orio F Jr, Russo T, Falbo A, Tolino A, Lombardi G, Cimini V and Zullo F: Antiproliferativeand proapoptotic effects of raloxifene on uterine leiomyomas in postmenopausal women. Fertil Steril. 84:154–161. 2005. View Article : Google Scholar : PubMed/NCBI
41	Tsurimoto T: PCNA, a multifunctional ring on DNA. Biochim Biophys Acta. 1443:23–39. 1998. View Article : Google Scholar : PubMed/NCBI
42	Majka J and Burgers PM: The PCNA-RFC families of DNA clamps and clamp loaders. Prog Nucleic Acid Res Mol Biol. 78:227–260. 2004. View Article : Google Scholar : PubMed/NCBI
43	Petros AM, Olejniczak ET and Fesik SW: Structural biology of the Bcl-2 family of proteins. Biochim Biophys Acta. 1644:83–94. 2004. View Article : Google Scholar : PubMed/NCBI
44	Sorenson CM: Bcl-2 family members and disease. Biochim Biophys Acta. 1644:169–177. 2004. View Article : Google Scholar : PubMed/NCBI
45	Cory S, Huang DC and Adams JM: The Bcl-2 family: Roles in cell survival andoncogenesis. Oncogene. 22:8590–8607. 2003. View Article : Google Scholar : PubMed/NCBI
46	Burroughs KD, Fuchs-Young R, Davis B and Walker CL: Altered hormonalresponsiveness of proliferation and apoptosis during myometrial maturation and the development of uterine leiomyomas in the rat. Biol Reprod. 63:1322–1330. 2000. View Article : Google Scholar : PubMed/NCBI
47	Martel KM, Ko AC, Christman GM and Stribley JM: Apoptosis in human uterine leiomyomas. Semin Reprod Med. 22:91–103. 2004. View Article : Google Scholar : PubMed/NCBI
48	Maruo T, Ohara N, Wang J and Matsuo H: Sex steroidal regulation of uterine leiomyoma growth and apoptosis. Hum Reprod Update. 10:207–220. 2004. View Article : Google Scholar : PubMed/NCBI
49	Kao AP, Wang KH, Long CY, Chai CY, Tsai CF, Hsieh TH, Hsu CY, Chang CC, Lee JN and Tsai EM: Interleukin-1β induces cyclooxygenase-2 expression and promotes theinvasive ability of human mesenchymal stem cells derived from ovarianendometrioma. Fertil Steril. 96:678–684.e1. 2011. View Article : Google Scholar : PubMed/NCBI
50	Dairkee SH, Seok J, Champion S, Sayeed A, Mindrinos M, Xiao W, Davis RW and Goodson WH: Biphenol A induces a profile of tumor aggressiveness in high-risk cellsfrom breast cancer patients. Cancer Res. 68:2076–2080. 2008. View Article : Google Scholar : PubMed/NCBI
51	Wang KH, Kao AP, Chang CC, Lee JN, Chai CY, Hou MF, Liu CM and Tsai EM: Modulation of tumorigenesis and oestrogen receptor-alpha expression by cell culture conditions in a stem cell-derived breast epithelial cell line. Biol Cell. 102:159–172. 2010. View Article : Google Scholar : PubMed/NCBI