Establishment and characterization of cell lines derived from complete hydatidiform mole

Yamamoto,Eiko; Niimi,Kaoru; Kiyono,Tohru; Yamamoto,Toshimichi; Nishino,Kimihiro; Nakamura,Kenichi; Kotani,Tomomi; Kajiyama,Hiroaki; Shibata,Kiyosumi; Kikkawa,Fumitaka

doi:10.3892/ijmm.2017.3067

Establishment and characterization of cell lines derived from complete hydatidiform mole

Authors:
- Eiko Yamamoto
- Kaoru Niimi
- Tohru Kiyono
- Toshimichi Yamamoto
- Kimihiro Nishino
- Kenichi Nakamura
- Tomomi Kotani
- Hiroaki Kajiyama
- Kiyosumi Shibata
- Fumitaka Kikkawa
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Affiliations: Department of Obstetrics and Gynecology, Nagoya University Graduate School of Medicine, Nagoya 466-8550, Japan, Division of Carcinogenesis and Cancer Prevention, National Cancer Center Research Institute, Tokyo 104-0045, Japan, Department of Legal Medicine and Bioethics, Nagoya University Graduate School of Medicine, Nagoya 466-8550, Japan
Published online on: July 14, 2017 https://doi.org/10.3892/ijmm.2017.3067
Pages: 614-622
Copyright: © Yamamoto et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Gestational trophoblastic diseases (GTDs) are a group of diseases characterized by abnormal cellular proliferation of atypical trophoblasts. A hydatidiform mole is an abnormal pregnancy caused by genetic fertilization disorders, and it can be classified as a complete hydatidiform mole (CHM) or a partial hydatidiform mole. The aim of this study was to establish cell lines from CHMs and to characterize the cells for future studies concerning GTD. HMol1-2C, HMol1-3B, HMol1-8 and HMol3-1B were established from primary cultures of CHM explants following the introduction of different combinations of genes including human telomerase reverse transcriptase (hTERT), a mutant form of CDK (CDK4R24C), cyclin D1, p53C234, MYC and HRAS. HMol1-2C, HMol1-3B, and HMol3-1B were confirmed to originate from trophoblasts of androgenic, homozygous CHMs. These three cell lines exhibited low human chorionic gonadotropin secretion, low migration and invasion abilities, and the potential to differentiate into syncytiotrophoblastic cells via forskolin treatment. These results suggest that these cells exhibit characteristics of trophoblastic cells, especially cytotrophoblastic cells. HMol1-8 was found to consist of diploid cells and originated from maternal cells, suggesting that they were derived from decidual cells. In conclusion, we successfully established three cell lines from CHMs by introduction of hTERT and other genes. Analysis revealed that the genetic origin of each cell line was identical with that of the original molar tissue, and the cell lines exhibited characteristics of trophoblastic cells, which are similar to undifferentiated cytotrophoblasts.

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1	Kajii T and Ohama K: Androgenetic origin of hydatidiform mole. Nature. 268:633–634. 1977. View Article : Google Scholar : PubMed/NCBI
2	Wake N, Seki T, Fujita H, Okubo H, Sakai K, Okuyama K, Hayashi H, Shiina Y, Sato H, Kuroda M, et al: Malignant potential of homozygous and heterozygous complete moles. Cancer Res. 44:1226–1230. 1984.PubMed/NCBI
3	Vejerslev LO, Dissing J, Hansen HE and Poulsen H: Hydatidiform mole: Genetic origin in polyploid conceptuses. Hum Genet. 76:11–19. 1987. View Article : Google Scholar : PubMed/NCBI
4	Ohama K, Ueda K, Okamoto E, Takenaka M and Fujiwara A: Cytogenetic and clinicopathologic studies of partial moles. Obstet Gynecol. 68:259–262. 1986.PubMed/NCBI
5	Kaneki E, Kobayashi H, Hirakawa T, Matsuda T, Kato H and Wake N: Incidence of postmolar gestational trophoblastic disease in androgenetic moles and the morphological features associated with low risk postmolar gestational trophoblastic disease. Cancer Sci. 101:1717–1721. 2010. View Article : Google Scholar : PubMed/NCBI
6	Seckl MJ, Sebire NJ and Berkowitz RS: Gestational trophoblastic disease. Lancet. 376:717–729. 2010. View Article : Google Scholar : PubMed/NCBI
7	Kan M, Yamamoto E, Niimi K, Tamakoshi K, Sekiya Y, Nishino K, Ino K and Kikkawa F: Gestational trophoblastic neoplasia and pregnancy outcome after routine second curettage for hydatidiform mole: A retrospective observational study. J Reprod Med. 61:373–379. 2016.
8	Taillon-Miller P, Bauer-Sardiña I, Zakeri H, Hillier L, Mutch DG and Kwok PY: The homozygous complete hydatidiform mole: A unique resource for genome studies. Genomics. 46:307–310. 1997. View Article : Google Scholar
9	Steinberg KM, Schneider VA, Graves-Lindsay TA, Fulton RS, Agarwala R, Huddleston J, Shiryev SA, Morgulis A, Surti U, Warren WC, et al: Single haplotype assembly of the human genome from a hydatidiform mole. Genome Res. 24:2066–2076. 2014. View Article : Google Scholar : PubMed/NCBI
10	Kuroda K, Kiyono T, Eitsuka T, Isogai H, Takahashi K, Donai K, Isogai E and Fukuda T: Establishment of cell lines derived from the genus Macaca through controlled expression of cell cycle regulators. J Cell Biochem. 116:205–211. 2015. View Article : Google Scholar
11	Donai K, Kiyono T, Eitsuka T, Guo Y, Kuroda K, Sone H, Isogai E and Fukuda T: Bovine and porcine fibroblasts can be immortalized with intact karyotype by the expression of mutant cyclin dependent kinase 4, cyclin D, and telomerase. J Biotechnol. 176:50–57. 2014. View Article : Google Scholar : PubMed/NCBI
12	Sasaki R, Narisawa-Saito M, Yugawa T, Fujita M, Tashiro H, Katabuchi H and Kiyono T: Oncogenic transformation of human ovarian surface epithelial cells with defined cellular oncogenes. Carcinogenesis. 30:423–431. 2009. View Article : Google Scholar : PubMed/NCBI
13	Okamoto T, Aoyama T, Nakayama T, Nakamata T, Hosaka T, Nishijo K, Nakamura T, Kiyono T and Toguchida J: Clonal heterogeneity in differentiation potential of immortalized human mesenchymal stem cells. Biochem Biophys Res Commun. 295:354–361. 2002. View Article : Google Scholar : PubMed/NCBI
14	Bayasula IA, Iwase A, Kiyono T, Takikawa S, Goto M, Nakamura T, Nagatomo Y, Nakahara T, Kotani T, Kobayashi H, et al: Establishment of a human nonluteinized granulosa cell line that transitions from the gonadotropin-independent to the gonadotropin-dependent status. Endocrinology. 153:2851–2860. 2012. View Article : Google Scholar : PubMed/NCBI
15	Carey BW, Markoulaki S, Hanna J, Saha K, Gao Q, Mitalipova M and Jaenisch R: Reprogramming of murine and human somatic cells using a single polycistronic vector. Proc Natl Acad Sci USA. 106:157–162. 2009. View Article : Google Scholar :
16	Miyoshi H, Blömer U, Takahashi M, Gage FH and Verma IM: Development of a self-inactivating lentivirus vector. J Virol. 72:8150–8157. 1998.PubMed/NCBI
17	Yamamoto E, Ito T, Abe A, Sido F, Ino K, Itakura A, Mizutani S, Dovat S, Nomura S and Kikkawa F: Ikaros is expressed in human extravillous trophoblasts and involved in their migration and invasion. Mol Hum Reprod. 11:825–831. 2005. View Article : Google Scholar : PubMed/NCBI
18	Yamamoto E, Ino K, Miyoshi E, Inamori K, Abe A, Sumigama S, Iwase A, Kajiyama H, Shibata K, Nawa A, et al: N-acetylglucosaminyltransferase V regulates extravillous trophoblast invasion through glycosylation of alpha5beta1 integrin. Endocrinology. 150:990–999. 2009. View Article : Google Scholar
19	Austgulen R, Chedwick L, Vogt Isaksen C, Vatten L and Craven C: Trophoblast apoptosis in human placenta at term as detected by expression of a cytokeratin 18 degradation product of caspase. Arch Pathol Lab Med. 126:1480–1486. 2002.PubMed/NCBI
20	Sasagawa M, Yamazaki T, Endo M, Kanazawa K and Takeuchi S: Immunohistochemical localization of HLA antigens and placental proteins (alpha hCG, beta hCG CTP, hPL and SP1 in villous and extravillous trophoblast in normal human pregnancy: A distinctive pathway of differentiation of extravillous trophoblast. Placenta. 8:515–528. 1987. View Article : Google Scholar : PubMed/NCBI
21	Lawler SD, Fisher RA and Dent J: A prospective genetic study of complete and partial hydatidiform moles. Am J Obstet Gynecol. 164:1270–1277. 1991. View Article : Google Scholar : PubMed/NCBI
22	Devergne O, Coulomb-L'Herminé A, Capel F, Moussa M and Capron F: Expression of Epstein-Barr virus-induced gene 3, an interleukin-12 p40-related molecule, throughout human pregnancy: Involvement of syncytiotrophoblasts and extravillous trophoblasts. Am J Pathol. 159:1763–1776. 2001. View Article : Google Scholar : PubMed/NCBI
23	Niimi K, Yamamoto E, Fujiwara S, Shinjo K, Kotani T, Umezu T, Kajiyama H, Shibata K, Ino K and Kikkawa F: High expression of N-acetylglucosaminyltransferase IVa promotes invasion of choriocarcinoma. Br J Cancer. 107:1969–1977. 2012. View Article : Google Scholar : PubMed/NCBI
24	Zhu H, Hou CC, Luo LF, Hu YJ and Yang WX: Endometrial stromal cells and decidualized stromal cells: Origins, transformation and functions. Gene. 551:1–14. 2014. View Article : Google Scholar : PubMed/NCBI
25	Richards RG, Brar AK, Frank GR, Hartman SM and Jikihara H: Fibroblast cells from term human decidua closely resemble endometrial stromal cells: Induction of prolactin and insulin-like growth factor binding protein-1 expression. Biol Reprod. 52:609–615. 1995. View Article : Google Scholar : PubMed/NCBI
26	Telgmann R and Gellersen B: Marker genes of decidualization: Activation of the decidual prolactin gene. Hum Reprod Update. 4:472–479. 1998. View Article : Google Scholar
27	Dunn CL, Kelly RW and Critchley HO: Decidualization of the human endometrial stromal cell: An enigmatic transformation. Reprod Biomed Online. 7:151–161. 2003. View Article : Google Scholar : PubMed/NCBI
28	Kim JJ, Jaffe RC and Fazleabas AT: Insulin-like growth factor binding protein-1 expression in baboon endometrial stromal cells: Regulation by filamentous actin and requirement for de novo protein synthesis. Endocrinology. 140:997–1004. 1999. View Article : Google Scholar : PubMed/NCBI
29	Lockwood CJ, Krikun G, Caze R, Rahman M, Buchwalder LF and Schatz F: Decidual cell-expressed tissue factor in human pregnancy and its involvement in hemostasis and preeclampsia-related angiogenesis. Ann NY Acad Sci. 1127:67–72. 2008. View Article : Google Scholar : PubMed/NCBI
30	Graham CH, Hawley TS, Hawley RG, MacDougall JR, Kerbel RS, Khoo N and Lala PK: Establishment and characterization of first trimester human trophoblast cells with extended lifespan. Exp Cell Res. 206:204–211. 1993. View Article : Google Scholar : PubMed/NCBI
31	Wang YL, Qiu W, Feng HC, Li YX, Zhuang LZ, Wang Z, Liu Y, Zhou JQ, Zhang DH and Tsao GS: Immortalization of normal human cytotrophoblast cells by reconstitution of telomeric reverse transcriptase activity. Mol Hum Reprod. 12:451–460. 2006. View Article : Google Scholar : PubMed/NCBI
32	Li RH and Zhuang LZ: The effects of growth factors on human normal placental cytotrophoblast cell proliferation. Hum Reprod. 12:830–834. 1997. View Article : Google Scholar : PubMed/NCBI
33	Katayama M, Kiyono T, Horie K, Hirayama T, Eitsuka T, Kuroda K, Donai K, Hidema S, Nishimori K and Fukuda T: Establishment of an immortalized cell line derived from the prairie vole via lentivirus-mediated transduction of mutant cyclin-dependent kinase 4, cyclin D, and telomerase reverse transcriptase. Exp Anim. 65:87–96. 2016. View Article : Google Scholar :
34	Kuroda K, Kiyono T, Isogai E, Masuda M, Narita M, Okuno K and Koyanagi Y: Fukuda T. Immortalization of fetal bovine colon epithelial cells by expression of human cyclin D1, mutant cyclin dependent kinase 4, and telomerase reverse transcriptase: An in vitro model for bacterial infection. PLoS One. 10:e01434732015. View Article : Google Scholar : PubMed/NCBI
35	Inagawa Y, Yamada K, Yugawa T, Ohno S, Hiraoka N, Esaki M, Shibata T, Aoki K, Saya H and Kiyono T: A human cancer xenograft model utilizing normal pancreatic duct epithelial cells conditionally transformed with defined oncogenes. Carcinogenesis. 35:1840–1846. 2014. View Article : Google Scholar : PubMed/NCBI
36	Sundvall L, Lund H, Niemann I, Jensen UB, Bolund L and Sunde L: Tetraploidy in hydatidiform moles. Hum Reprod. 28:2010–2020. 2013. View Article : Google Scholar : PubMed/NCBI
37	Zygmunt M, Hahn D, Münstedt K, Bischof P and Lang U: Invasion of cytotrophoblastic JEG-3 cells is stimulated by hCG in vitro. Placenta. 19:587–593. 1998. View Article : Google Scholar : PubMed/NCBI