Overexpression of satellite alpha transcripts leads to chromosomal instability via segregation errors at specific chromosomes

Ichida,Kosuke; Suzuki,Koichi; Fukui,Taro; Takayama,Yuji; Kakizawa,Nao; Watanabe,Fumiaki; Ishikawa,Hideki; Muto,Yuta; Kato,Takaharu; Saito,Masaaki; Futsuhara,Kazushige; Miyakura,Yasuyuki; Noda,Hiroshi; Ohmori,Tsukasa; Konishi,Fumio; Rikiyama,Toshiki

doi:10.3892/ijo.2018.4321

Overexpression of satellite alpha transcripts leads to chromosomal instability via segregation errors at specific chromosomes

Authors:
- Kosuke Ichida
- Koichi Suzuki
- Taro Fukui
- Yuji Takayama
- Nao Kakizawa
- Fumiaki Watanabe
- Hideki Ishikawa
- Yuta Muto
- Takaharu Kato
- Masaaki Saito
- Kazushige Futsuhara
- Yasuyuki Miyakura
- Hiroshi Noda
- Tsukasa Ohmori
- Fumio Konishi
- Toshiki Rikiyama
View Affiliations

Affiliations: Department of Surgery, Saitama Medical Center, Jichi Medical University, Saitama-shi, Saitama 330-8503, Japan, Department of Biochemistry, Jichi Medical University, Shimotsuke-shi, Tochigi 329-0498, Japan, Nerima-Hikarigaoka Hospital, Tokyo 179-0072, Japan
Published online on: March 16, 2018 https://doi.org/10.3892/ijo.2018.4321
Pages: 1685-1693

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

The impairment of the stability of the chromosomal structure facilitates the abnormal segregation of chromosomes, thus increasing the risk of carcinogenesis. Chromosomal stability during segregation is managed by appropriate methylation at the centromere of chromosomes. Insufficient methylation, or hypomethylation, results in chromosomal instability. The centromere consists of satellite alpha repetitive sequences, which are ideal targets for DNA hypomethylation, resulting in the overexpression of satellite alpha transcript (SAT). The overexpression of SAT has been reported to induce the abnormal segregation of chromosomes. In this study, we verified the oncogenic pathway via chromosomal instability involving DNA hypomethylation and the overexpression of SAT. For this purpose, we constructed lentiviral vectors expressing SAT and control viruses and then infected human mammary epithelial cells with these vectors. The copy number alterations and segregation errors of chromosomes were evaluated by microarray-based comparative genomic hybridization (array CGH) and immunocytochemistry, respectively. The levels of hypomethylation of satellite alpha sequences were determined by MethyLight polymerase chain reaction. Clinical specimens from 45 patients with breast cancer were recruited to verify the data in vitro. The results of immunocytochemistry revealed that the incidence of segregation errors was significantly higher in the cells overexpressing SAT than in the controls. An array CGH identified the specific chromosomes of 8q and 20q as frequent sites of copy number alterations in cells with SAT overexpression, although no such sites were noted in the controls, which was consistent with the data from clinical specimens. A regression analysis revealed that the expression of SAT was significantly associated with the levels of hypomethylation of satellite alpha sequences. On the whole, the overexpression of SAT led to chromosomal instability via segregation errors at specific chromosomes in connection with DNA hypomethylation, which was also recognized in clinical specimens of patients with breast cancer. Thus, this oncogenic pathway may be involved in the development of breast cancer.

View Figures

View References

1	Feinberg AP and Vogelstein B: Hypomethylation distinguishes genes of some human cancers from their normal counterparts. Nature. 301:89–92. 1983. View Article : Google Scholar : PubMed/NCBI
2	Rodriguez J, Frigola J, Vendrell E, Risques RA, Fraga MF, Morales C, Moreno V, Esteller M, Capellà G, Ribas M, et al: Chromosomal instability correlates with genome-wide DNA demethylation in human primary colorectal cancers. Cancer Res. 66:8462–9468. 2006. View Article : Google Scholar : PubMed/NCBI
3	Herrera LA, Prada D, Andonegui MA and Dueñas-González A: The epigenetic origin of aneuploidy. Curr Genomics. 9:43–50. 2008. View Article : Google Scholar : PubMed/NCBI
4	Kawano H, Saeki H, Kitao H, Tsuda Y, Otsu H, Ando K, Ito S, Egashira A, Oki E, Morita M, et al: Chromosomal instability associated with global DNA hypomethylation is associated with the initiation and progression of esophageal squamous cell carcinoma. Ann Surg Oncol. 21(Suppl 4): S696–S702. 2014. View Article : Google Scholar : PubMed/NCBI
5	Esteller M, Silva JM, Dominguez G, Bonilla F, Matias-Guiu X, Lerma E, Bussaglia E, Prat J, Harkes IC, Repasky EA, et al: Promoter hypermethylation and BRCA1 inactivation in sporadic breast and ovarian tumors. J Natl Cancer Inst. 92:564–569. 2000. View Article : Google Scholar : PubMed/NCBI
6	Yang B, Guo M, Herman JG and Clark DP: Aberrant promoter methylation profiles of tumor suppressor genes in hepatocellular carcinoma. Am J Pathol. 163:1101–1107. 2003. View Article : Google Scholar : PubMed/NCBI
7	Herman JG and Baylin SB: Gene silencing in cancer in association with promoter hypermethylation. N Engl J Med. 349:2042–2054. 2003. View Article : Google Scholar : PubMed/NCBI
8	Itano O, Ueda M, Kikuchi K, Hashimoto O, Hayatsu S, Kawaguchi M, Seki H, Aiura K and Kitajima M: Correlation of postoperative recurrence in hepatocellular carcinoma with demethylation of repetitive sequences. Oncogene. 21:789–797. 2002. View Article : Google Scholar : PubMed/NCBI
9	Widschwendter M, Jiang G, Woods C, Müller HM, Fiegl H, Goebel G, Marth C, Müller-Holzner E, Zeimet AG, Laird PW, et al: DNA hypomethylation and ovarian cancer biology. Cancer Res. 64:4472–4480. 2004. View Article : Google Scholar : PubMed/NCBI
10	Igarashi S, Suzuki H, Niinuma T, Shimizu H, Nojima M, Iwaki H, Nobuoka T, Nishida T, Miyazaki Y, Takamaru H, et al: A novel correlation between LINE-1 hypomethylation and the malignancy of gastrointestinal stromal tumors. Clin Cancer Res. 16:5114–5123. 2010. View Article : Google Scholar : PubMed/NCBI
11	Eden A, Gaudet F, Waghmare A and Jaenisch R: Chromosomal instability and tumors promoted by DNA hypomethylation. Science. 300:4552003. View Article : Google Scholar
12	Gaudet F, Hodgson JG, Eden A, Jackson-Grusby L, Dausman J, Gray JW, Leonhardt H and Jaenisch R: Induction of tumors in mice by genomic hypomethylation. Science. 300:489–492. 2003. View Article : Google Scholar : PubMed/NCBI
13	Suzuki K, Suzuki I, Leodolter A, Alonso S, Horiuchi S, Yamashita K and Perucho M: Global DNA demethylation in gastrointestinal cancer is age dependent and precedes genomic damage. Cancer Cell. 9:199–207. 2006. View Article : Google Scholar : PubMed/NCBI
14	Saito M, Suzuki K, Maeda T, Kato T, Kamiyama H, Koizumi K, Miyaki Y, Okada S, Kiyozaki H and Konishi F: The accumulation of DNA demethylation in Sat α in normal gastric tissues with Helicobacter pylori infection renders susceptibility to gastric cancer in some individuals. Oncol Rep. 27:1717–1725. 2012.PubMed/NCBI
15	Plohl M, Meštrović N and Mravinac B: Centromere identity from the DNA point of view. Chromosoma. 123:313–325. 2014. View Article : Google Scholar : PubMed/NCBI
16	Ugarkovic D: Functional elements residing within satellite DNAs. EMBO Rep. 6:1035–1039. 2005. View Article : Google Scholar : PubMed/NCBI
17	Ideue T, Cho Y, Nishimura K and Tani T: Involvement of satellite I noncoding RNA in regulation of chromosome segregation. Genes Cells. 19:528–538. 2014. View Article : Google Scholar : PubMed/NCBI
18	Rošić S, Köhler F and Erhardt S: Repetitive centromeric satellite RNA is essential for kinetochore formation and cell division. J Cell Biol. 207:335–349. 2014. View Article : Google Scholar
19	McNulty SM, Sullivan LL and Sullivan BA: Human centromeres produce chromosome-specific and array-specific alpha satellite transcripts that are complexed with CENP-A and CENP-C. Dev Cell. 42:226–240.e6. 2017. View Article : Google Scholar : PubMed/NCBI
20	Ting DT, Lipson D, Paul S, Brannigan BW, Akhavanfard S, Coffman EJ, Contino G, Deshpande V, Iafrate AJ, Letovsky S, et al: Aberrant overexpression of satellite repeats in pancreatic and other epithelial cancers. Science. 331:593–596. 2011. View Article : Google Scholar : PubMed/NCBI
21	Zhu Q, Pao GM, Huynh AM, Suh H, Tonnu N, Nederlof PM, Gage FH and Verma IM: BRCA1 tumour suppression occurs via heterochromatin-mediated silencing. Nature. 477:179–184. 2011. View Article : Google Scholar : PubMed/NCBI
22	Kishikawa T, Otsuka M, Yoshikawa T, Ohno M, Ijichi H and Koike K: Satellite RNAs promote pancreatic oncogenic processes via the dysfunction of YBX1. Nat Commun. 7:130062016. View Article : Google Scholar : PubMed/NCBI
23	Lehnertz B, Ueda Y, Derijck AA, Braunschweig U, Perez-Burgos L, Kubicek S, Chen T, Li E, Jenuwein T and Peters AH: Suv39h-mediated histone H3 lysine 9 methylation directs DNA methylation to major satellite repeats at pericentric heterochromatin. Curr Biol. 13:1192–1200. 2003. View Article : Google Scholar : PubMed/NCBI
24	Bouzinba-Segard H, Guais A and Francastel C: Accumulation of small murine minor satellite transcripts leads to impaired centromeric architecture and function. Proc Natl Acad Sci USA. 103:8709–8714. 2006. View Article : Google Scholar : PubMed/NCBI
25	Eymery A, Horard B, El Atifi-Borel M, Fourel G, Berger F, Vitte AL, Van den Broeck A, Brambilla E, Fournier A, Callanan M, et al: A transcriptomic analysis of human centromeric and pericentric sequences in normal and tumor cells. Nucleic Acids Res. 37:6340–6354. 2009. View Article : Google Scholar : PubMed/NCBI
26	Leonova KI, Brodsky L, Lipchick B, Pal M, Novototskaya L, Chenchik AA, Sen GC, Komarova EA and Gudkov AV: P53 cooperates with DNA methylation and a suicidal interferon response to maintain epigenetic silencing of repeats and noncoding RNAs. Proc Natl Acad Sci USA. 110:E89–E98. 2013. View Article : Google Scholar :
27	Weisenberger DJ, Campan M, Long TI, Kim M, Woods C, Fiala E, Ehrlich M and Laird PW: Analysis of repetitive element DNA methylation by MethyLight. Nucleic Acids Res. 33:6823–6836. 2005. View Article : Google Scholar : PubMed/NCBI
28	Hagleitner MM, Lankester A, Maraschio P, Hultén M, Fryns JP, Schuetz C, Gimelli G, Davies EG, Gennery A, Belohradsky BH, et al: Clinical spectrum of immunodeficiency, centromeric instability and facial dysmorphism (ICF syndrome). J Med Genet. 45:93–99. 2008. View Article : Google Scholar
29	Walton EL, Francastel C and Velasco G: Dnmt3b prefers germ line genes and centromeric regions: Lessons from the ICF Syndrome and Cancer and Implications for Diseases. Biology (Basel). 3:578–605. 2014.
30	Tsuji K, Kawauchi S, Saito S, Furuya T, Ikemoto K, Nakao M, Yamamoto S, Oka M, Hirano T and Sasaki K: Breast cancer cell lines carry cell line-specific genomic alterations that are distinct from aberrations in breast cancer tissues: Comparison of the CGH profiles between cancer cell lines and primary cancer tissues. BMC Cancer. 10:152010. View Article : Google Scholar : PubMed/NCBI
31	Uchida M, Tsukamoto Y, Uchida T, Ishikawa Y, Nagai T, Hijiya N, Nguyen LT, Nakada C, Kuroda A, Okimoto T, et al: Genomic profiling of gastric carcinoma in situ and adenomas by array-based comparative genomic hybridization. J Pathol. 221:96–105. 2010. View Article : Google Scholar : PubMed/NCBI
32	Tabach Y, Kogan-Sakin I, Buganim Y, Solomon H, Goldfinger N, Hovland R, Ke XS, Oyan AM, Kalland KH, Rotter V, et al: Amplification of the 20q chromosomal arm occurs early in tumorigenic transformation and may initiate cancer. PLoS One. 6:e146322011. View Article : Google Scholar : PubMed/NCBI
33	Orsetti B, Selves J, Bascoul-Mollevi C, Lasorsa L, Gordien K, Bibeau F, Massemin B, Paraf F, Soubeyran I, Hostein I, et al: Impact of chromosomal instability on colorectal cancer progression and outcome. BMC Cancer. 14:1212014. View Article : Google Scholar : PubMed/NCBI