Low folate levels are associated with methylation-mediated transcriptional repression of miR-203 and miR-375 during cervical carcinogenesis

Hao,Min; Zhao,Weihong; Zhang,Lili; Wang,Honghong; Yang,Xin

doi:10.3892/ol.2016.4449

Low folate levels are associated with methylation-mediated transcriptional repression of miR-203 and miR-375 during cervical carcinogenesis

Authors:
- Min Hao
- Weihong Zhao
- Lili Zhang
- Honghong Wang
- Xin Yang
View Affiliations

Affiliations: Department of Obstetrics and Gynecology, The Second Hospital of Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China
Published online on: April 18, 2016 https://doi.org/10.3892/ol.2016.4449
Pages: 3863-3869

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 aim of the present study was to investigate the correlation between a lack of folic acid and the abnormal expression of microRNA (miR)-203 and miR-375 in cervical cancer. In total, 60 tissue samples of cervical intraepithelial neoplasia (CIN) or stage IA‑IIA cervical cancer (study group), and 30 samples without soluble interleukin or malignancy (control group) were examined. The expression of miR‑203 and miR‑375 was detected using reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR), and the difference in expression levels was quantified using the 2‑ΔΔCq method. In addition, CaSki cervical cancer cells were cultured in vitro and treated with various concentrations of folic acid. The DNA methylation states of miR‑203 and miR‑375 were subsequently detected by methylation‑specific PCR, and the expression levels were evaluated using RT‑PCR. miR‑203 and miR‑375 were significantly downregulated in CIN and cervical cancer tissues, compared with the control group. There was a marked difference in terms of the expression levels of miR‑375 between the two groups (P<0.05). In CaSki cells, as the concentration of folic acid increased, the positive rate of DNA methylation of miR‑203 and miR‑375 decreased, while the expression levels of miR‑203 and miR‑375 demonstrated a gradual increase, which indicated that the latter two parameters were negatively correlated (P<0.05). Compared with normal cervical tissue, the expression levels of miR‑203 and miR‑375 were downregulated in CIN and cervical cancer. Methylation of these two miRs was apparent in CaSki cells, and was associated with a lack of folic acid. Therefore, reduced levels of folic acid, leading to increased methylation of miR‑203 and miR-375, may be significant events during cervical carcinogenesis.

View Figures

View References

1	Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D and Bray F: Cancer incidence and mortality worldwide: Sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer. 136:E359–E386. 2015. View Article : Google Scholar : PubMed/NCBI
2	Schiffman MH and Castle P: Epidemiologic studies of a necessary causal risk factor: Human papillomavirus infection and cervical neoplasia. J Natl Cancer Inst. 95:E22003. View Article : Google Scholar : PubMed/NCBI
3	Choi YJ and Park JS: Clinical significance of human papillomavirus genotyping. J Gynecol Oncol. 27:e212016. View Article : Google Scholar : PubMed/NCBI
4	Ho GY, Bierman R, Beardsley L, Chang CJ and Burk RD: Natural history of cervicovaginal papillomavirus infection in young women. N Engl J Med. 338:423–428. 1998. View Article : Google Scholar : PubMed/NCBI
5	Zhao WH, Hao M, Cheng XT, Yang X, Wang ZL, Cheng KY, Liu FL and Bai YX: c-myc gene copy number variation in cervical exfoliated cells detected on fluorescence in situ hybridization for cervical cancer screening. Gynecol Obstet Invest. Jan 26–2016.(Epub ahead of print). View Article : Google Scholar : PubMed/NCBI
6	Wilting SM, Snijders PJ, Verlaat W, Jaspers A, van de Wiel MA, van Wieringen WN, Meijer GA, Kenter GG, Yi Y, le Sage C, et al: Altered microRNA expression associated with chromosomal changes contributes to cervical carcinogenesis. Oncogene. 32:106–116. 2013. View Article : Google Scholar : PubMed/NCBI
7	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
8	Wang L, Yue Y, Wang X and Jin H: Function and clinical potential of microRNAs in hepatocellular carcinoma. Oncol Lett. 10:3345–3353. 2015.PubMed/NCBI
9	Lo R and Weksberg R: Biological and biochemical modulation of DNA methylation. Epigenomics. 6:593–602. 2014. View Article : Google Scholar : PubMed/NCBI
10	Sarkar D, Leung EY, Baguley BC, Finlay GJ and Askarian-Amiri ME: Epigenetic regulation in human melanoma: Past and future. Epigenetics. 10:103–121. 2015. View Article : Google Scholar : PubMed/NCBI
11	Wang LQ and Chim CS: DNA methylation of tumor-suppressor miRNA genes in chronic lymphocytic leukemia. Epigenomics. 7:461–473. 2015. View Article : Google Scholar : PubMed/NCBI
12	Ferlay J, Shin HR, Bray F, Forman D, Mathers C and Parkin DM: Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 127:2893–2917. 2010. View Article : Google Scholar : PubMed/NCBI
13	Wang JT, Ding L, Jiang SW, Hao J, Zhao WM, Zhou Q, Yang ZK and Zhang L: Folate deficiency and aberrant expression of DNA methyltransferase 1 were associated with cervical cancerization. Curr Pharm Des. 20:1639–1646. 2014. View Article : Google Scholar : PubMed/NCBI
14	Flatley JE, McNeir K, Balasubramani L, Tidy J, Stuart EL, Young TA and Powers HJ: Folate status and aberrant DNA methylation are associated with HPV infection and cervical pathogenesis. Cancer Epidemiol Biomarkers Prev. 18:2782–2789. 2009. View Article : Google Scholar : PubMed/NCBI
15	Flatley JE, Sargent A, Kitchener HC, Russell JM and Powers HJ: Tumour suppressor gene methylation and cervical cell folate concentration are determinants of high-risk human papillomavirus persistence: A nested case control study. BMC Cancer. 14:8032014. View Article : Google Scholar : PubMed/NCBI
16	Bai LX, Wang JT, Ding L, Jiang SW, Kang HJ, Gao CF, Chen X, Chen C and Zhou Q: Folate deficiency and FHIT hypermethylation and HPV 16 infection promote cervical cancerization. Asian Pac J Cancer Prev. 15:9313–9317. 2014. View Article : Google Scholar : PubMed/NCBI
17	Lewin SN, Herzog TJ, Barrena Medel NI, Deutsch I, Burke WM, Sun X and Wright JD: Comparative performance of the 2009 International Federation of Gynecology and Obstetrics' staging system for uterine corpus cancer. Obstet Gynecol. 116:1141–1149. 2010. View Article : Google Scholar : PubMed/NCBI
18	Liu HQ, Wang YH, Wang LL and Hao M: P16INK4A and survivin: Diagnostic and prognostic markers in cervical intraepithelial neoplasia and cervical squamous cell carcinoma. Exp Mol Pathol. 99:44–49. 2015. View Article : Google Scholar : PubMed/NCBI
19	Wright TC, Ronnett BM, Kurman RJ and Ferenczy A: Precancerous lesions of the cervix. Kurman RJ, Ellenson LH and Ronnett BM: Blaustein's Pathology of the Female Genital Tract (6th). Springer US. (New York, NY). 193–252. 2011. View Article : Google Scholar
20	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
21	Xiao W, Zhao H, Dong W, Li Q, Zhu J, Li G, Zhang S and Ye M: Quantitative detection of methylated NDRG4 gene as a candidate biomarker for diagnosis of colorectal cancer. Oncol Lett. 9:1383–1387. 2015.PubMed/NCBI
22	Shi JF, Canfell K, Lew JB and Qiao YL: The burden of cervical cancer in China: Synthesis of the evidence. Int J Cancer. 130:641–652. 2012. View Article : Google Scholar : PubMed/NCBI
23	Wang JT, Ma XC, Cheng YY, Ding L and Zhou Q: A case-control study on the association between folate and cervical cancer. Zhonghua Liu Xing Bing Xue Za Zhi. 27:424–427. 2006.(In Chinese). PubMed/NCBI
24	Cheung TH, Man KN, Yu MY, Yim SF, Siu NS, Lo KW, Doran G, Wong RR, Wang VW, Smith DI, et al: Dysregulated microRNAs in the pathogenesis and progression of cervical neoplasm. Cell Cycle. 11:2876–2884. 2012. View Article : Google Scholar : PubMed/NCBI
25	Jiménez-Wences H, Peralta-Zaragoza O and Fernández-Tilapa G: Human papilloma virus, DNA methylation and microRNA expression in cervical cancer (Review). Oncol Rep. 31:2467–2476. 2014.PubMed/NCBI
26	Yao T, Rao Q, Liu L, Zheng C, Xie Q, Liang J and Lin Z: Exploration of tumor-suppressive microRNAs silenced by DNA hypermethylation in cervical cancer. Virol J. 10:1752013. View Article : Google Scholar : PubMed/NCBI
27	Michel CI and Malumbres M: microRNA-203: Tumor suppression and beyond. Microrna. 2:118–126. 2013. View Article : Google Scholar : PubMed/NCBI
28	Chen T, Xu C, Chen J, Ding C, Xu Z, Li C and Zhao J: MicroRNA-203 inhibits cellular proliferation and invasion by targeting Bmi1 in non-small cell lung cancer. Oncol Lett. 9:2639–2646. 2015.PubMed/NCBI
29	Gocze K, Gombos K, Juhasz K, Kovacs K, Kajtar B, Benczik M, Gocze P, Patczai B, Arany I and Ember I: Unique microRNA expression profiles in cervical cancer. Anticancer Res. 33:2561–2567. 2013.PubMed/NCBI
30	Gocze K, Gombos K, Kovacs K, Juhasz K, Gocze P and Kiss I: MicroRNA expressions in HPV-induced cervical dysplasia and cancer. Anticancer Res. 35:523–530. 2015.PubMed/NCBI
31	Melar-New M and Laimins LA: Human papillomaviruses modulate expression of microRNA 203 upon epithelial differentiation to control levels of p63 proteins. J Virol. 84:5212–5221. 2010. View Article : Google Scholar : PubMed/NCBI
32	Greco D, Kivi N, Qian K, Leivonen SK, Auvinen P and Auvinen E: Human papillomavirus 16 E5 modulates the expression of host microRNAs. PLoS One. 6:e216462011. View Article : Google Scholar : PubMed/NCBI
33	Costa-Pinheiro P, Ramalho-Carvalho J, Vieira FQ, Torres-Ferreira J, Oliveira J, Gonçalves CS, Costa BM, Henrique R and Jerónimo C: MicroRNA-375 plays a dual role in prostate carcinogenesis. Clin Epigenetics. 7:422015. View Article : Google Scholar : PubMed/NCBI
34	Wang F, Li Y, Zhou J, Xu J, Peng C, Ye F, Shen Y, Lu W, Wan X and Xie X: miR-375 is down-regulated in squamous cervical cancer and inhibits cell migration and invasion via targeting transcription factor SP1. Am J Pathol. 179:2580–2588. 2011. View Article : Google Scholar : PubMed/NCBI
35	Shen Y, Wang P, Li Y, Ye F, Wang F, Wan X, Cheng X, Lu W and Xie X: miR-375 is upregulated in acquired paclitaxel resistance in cervical cancer. Br J Cancer. 109:92–99. 2013. View Article : Google Scholar : PubMed/NCBI
36	Shen Y, Zhou J, Li Y, Ye F, Wan X, Lu W, Xie X and Cheng X: miR-375 mediated acquired chemo-resistance in cervical cancer by facilitating EMT. PLoS One. 9:e1092992014. View Article : Google Scholar : PubMed/NCBI
37	Adams BD, Kasinski AL and Slack FJ: Aberrant regulation and function of microRNAs in cancer. Curr Biol. 24:R762–776. 2014. View Article : Google Scholar : PubMed/NCBI
38	Wilting SM, Verlaat W, Jaspers A, Makazaji NA, Agami R, Meijer CJ, Snijders PJ and Steenbergen RD: Methylation-mediated transcriptional repression of microRNAs during cervical carcinogenesis. Epigenetics. 8:220–228. 2013. View Article : Google Scholar : PubMed/NCBI
39	Geng Y, Gao R, Chen X, Liu X, Liao X, Li Y, Liu S, Ding Y, Wang Y and He J: Folate deficiency impairs decidualization and alters methylation patterns of the genome in mice. Mol Hum Reprod. 21:844–856. 2015. View Article : Google Scholar : PubMed/NCBI
40	Ho E, Beaver LM, Williams DE and Dashwood RH: Dietary factors and epigenetic regulation for prostate cancer prevention. Adv Nutr. 2:497–510. 2011. View Article : Google Scholar : PubMed/NCBI