Impact of gastro-oesophageal reflux on microRNA expression, location and function

BMC Gastroenterology

Damian J Hussey¹  Richard Hummel³  David St J Astill²  Grace Tan³  David I Watson¹  Michael Z Michael¹  Cameron M Smith³ 

[1] Flinders Centre for Cancer Prevention and Control, Flinders University, Bedford Park, South Australia, Australia;Department of Anatomical Pathology, Flinders Medical Centre, Bedford Park, South Australia, 5042, Australia;Department of Surgery, Flinders University, Room 3D213, Flinders Medical Centre, Bedford Park, South Australia, 5042, Australia

关键词: Barrett’s oesophagus;    Proliferation;    Apoptosis;    Ulcerative oesophagitis;    Gastro-oesophageal reflux disease;    microRNA;   

Others  :  858246
DOI  :  10.1186/1471-230X-13-4

 received in 2012-08-18, accepted in 2013-01-04,  发布年份 2013

【 摘 要 】

Background

Ulceration of the oesophageal squamous mucosa (ulcerative oesophagitis) is a pathological manifestation of gastro-oesophageal reflux disease, and is a major risk factor for the development of Barrett’s oesophagus. Barrett’s oesophagus is characterised by replacement of reflux-damaged oesophageal squamous epithelium with a columnar intestinal-like epithelium. We previously reported discovery of microRNAs that are differentially expressed between oesophageal squamous mucosa and Barrett’s oesophagus mucosa. Now, to better understand early steps in the initiation of Barrett’s oesophagus, we assessed the expression, location and function of these microRNAs in oesophageal squamous mucosa from individuals with ulcerative oesophagitis.

Methods

Quantitative real-time PCR was used to compare miR-21, 143, 145, 194, 203, 205 and 215 expression levels in oesophageal mucosa from individuals without pathological gastro-oesophageal reflux to individuals with ulcerative oesophagitis. Correlations between microRNA expression and messenger RNA differentiation markers BMP-4, CK8 and CK14 were analyzed. The cellular localisation of microRNAs within the oesophageal mucosa was determined using in-situ hybridisation. microRNA involvement in proliferation and apoptosis was assessed following transfection of a human squamous oesophageal mucosal cell line (Het-1A).

Results

miR-143, miR-145 and miR-205 levels were significantly higher in gastro-oesophageal reflux compared with controls. Elevated miR-143 expression correlated with BMP-4 and CK8 expression, and elevated miR-205 expression correlated negatively with CK14 expression. Endogenous miR-143, miR-145 and miR-205 expression was localised to the basal layer of the oesophageal epithelium. Transfection of miR-143, 145 and 205 mimics into Het-1A cells resulted in increased apoptosis and decreased proliferation.

Conclusions

Elevated miR-143, miR-145 and miR-205 expression was observed in oesophageal squamous mucosa of individuals with ulcerative oesophagitis. These miRNAs localised to the basal layer of the oesophageal epithelium. They reduced proliferation and increased apoptosis, and may play roles in regulating epithelial restoration in response to injury caused by gastro-oesophageal reflux.

【 授权许可】

   
2013 Smith et al.; licensee BioMed Central Ltd.

【 图 表 】

【 参考文献 】

• [1]van Roon AH, Mayne GC, Wijnhoven BP, et al.: Impact of gastro-oesophageal reflux on mucin mRNA expression in the oesophageal mucosa. J Gastrointest Surg 2008, 12:1331-1340.
• [2]Dixon J, Strugala V, Griffin SM, et al.: Oesophageal mucin: an adherent mucus gel barrier is absent in the normal oesophagus but present in columnar-lined Barrett's oesophagus. Am J Gastroenterol 2001, 96:2575-2583.
• [3]Mulder DJ, Pacheco I, Hurlbut DJ, et al.: FGF9-induced proliferative response to eosinophilic inflammation in oesophagitis. Gut 2009, 58:166-173.
• [4]Wetscher GJ, Schwelberger H, Unger A, et al.: Reflux-induced apoptosis of the oesophageal mucosa is inhibited in Barrett's epithelium. Am J Surg 1998, 176:569-573.
• [5]Moons LMG, Kusters JG, Van Delft JHM, et al.: A pro-inflammatory genotype predisposes to Barrett's oesophagus. Carcinogenesis 2008, 29:926-931.
• [6]Binato M, Fagundes R, Gurski R, Meurer L, Edelweiss MI: Immunohistochemical overexpression of the p53 protein and Ki-67 (MIB-1) antigen in patients with GERD and chronic oesophagitis. Appl Immunohistochem Mol Morphol 2010, 18:236-243.
• [7]Ronkainen J, Talley NJ, Storskrubb T, et al.: Erosive oesophagitis is a risk factor for Barrett's oesophagus: a community-based endoscopic follow-up study. Am J Gastroenterol 2011, 106:1946-1952.
• [8]Pera M, Cameron AJ, Trastek VF, Carpenter HA, Zinsmeister AR: Increasing incidence of adenocarcinoma of the oesophagus and oesophagogastric junction. Gastroenterology 1993, 104:510-513.
• [9]Milano F, van Baal JW, Buttar N, et al.: Bone morphogenetic protein 4 expressed in oesophagitis induces a columnar phenotype in oesophageal squamous cells. Gastroenterology 2007, 132:2412-2421.
• [10]Zhou G, Sun YG, Wang HB, et al.: Acid and bile salt up-regulate BMP4 expression in human oesophageal epithelium cells. Scand J Gastroenterol 2009, 44:926-932.
• [11]Pillai R, Bhattacharyya S, Filipowicz W: Repression of protein synthesis by miRNAs: how many mechanisms? Trends Cell Biol 2007, 17:118-126.
• [12]Dalmasso G, Thu Nguyen HT, Yan Y, et al.: MicroRNAs determine human intestinal epithelial cell fate. Differentiation 2010, 80:147-154.
• [13]Lena AM, Shalom-Feuerstein R, Di Val Cervo PR, et al.: miR-203 represses ‘stemness’ by repressing ΔNp63. Cell Death Differ 2008, 15:1187-1195.
• [14]Sotiropoulou G, Pampalakis G, Lianidou E, Mourelatos Z: Emerging roles of microRNAs as molecular switches in the integrated circuit of the cancer cell. RNA 2009, 15:1443-1461.
• [15]Wijnhoven BP, Hussey DJ, Watson DI, Tsykin A, Smith CM, Michael MZ: MicroRNA profiling of Barrett's oesophagus and oesophageal adenocarcinoma. Br J Surg 2010, 97:853-861.
• [16]Pena JTG, Sohn-Lee C, Rouhanifard SH, et al.: miRNA in situ hybridisation in formaldehyde and EDC–fixed tissues. Nat Methods 2009, 6:139-141.
• [17]Stoner GD, Kaighn ME, Reddel RR, et al.: Establishment and characterization of SV40 T-antigen immortalized human oesophageal epithelial cells. Cancer Res 1991, 51:365-371.
• [18]Simon P: Q-Gene: processing quantitative real-time RT-PCR data. Bioinformatics 2003, 19:1439-1440.
• [19]van Baal JW, Bozikas A, Pronk R, et al.: Cytokeratin and CDX-2 expression in Barrett's oesophagus. Scand J Gastroenterol 2008, 43:132-140.
• [20]Wu H, Zhu S, Mo YY: Suppression of cell growth and invasion by miR-205 in breast cancer. Cell Res 2009, 19:439-448.
• [21]Takagi T, Iio A, Nakagawa Y, Naoe T, Tanigawa N, Akao Y: Decreased expression of microRNA-143 and −145 in human gastric cancers. Oncology 2009, 77:12-21.
• [22]Akao Y, Nakagawa Y, Iio A, Naoe T: Role of microRNA-143 in Fas-mediated apoptosis in human T-cell leukemia Jurkat cells. Leuk Res 2009, 33:1530-1538.
• [23]Zaman MS, Chen Y, Deng G, et al.: The functional significance of microRNA-145 in prostate cancer. Br J Cancer 2010, 103:256-264.
• [24]Majid S, Dar AA, Saini S, et al.: MicroRNA-205-directed transcriptional activation of tumor suppressor genes in prostate cancer. Cancer 2010, 116:5637-5649.
• [25]van Baal JW, Verbeek RE, Bus P, et al.: microRNA-145 in Barrett's oesophagus: regulating BMP4 signalling via GATA6. Gut 2012. Epub ahead of print
• [26]Sempere L, Christensen M, Silahtaroglu A, et al.: Altered MicroRNA expression confined to specific epithelial cell subpopulations in breast cancer. Cancer Res 2007, 67:11612-11620.
• [27]Hwang HW, Wentzel EA, Mendell JT: A hexanucleotide element directs microRNA nuclear import. Science 2007, 315:97-100.
• [28]Liao JY, Ma LM, Guo YH, et al.: Deep sequencing of human nuclear and cytoplasmic small RNAs reveals an unexpectedly complex subcellular distribution of miRNAs and tRNA 3' trailers. PLoS One 2010, 5:e10563.
• [29]Taft RJ, Simons C, Nahkuri S, et al.: Nuclear-localised tiny RNAs are associated with transcription initiation and splice sites in metazoans. Nat Struct Mol Biol 2010, 17:1030-1034.
• [30]Caygill CP, Reed PI, Johnston BJ, Hill MJ, Ali MH, Levi S: A single centre's 20 years' experience of columnar-lined (Barrett's) oesophagus diagnosis. Eur J Gastroenterol Hepatol 1999, 11:1355-1358.