Induction of cancer-related microRNA expression profiling using excretory-secretory products of Clonorchis sinensis
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Clonorchis sinensis is a carcinogenic human liver fluke by which chronic infection is strongly associated with the development of cholangiocarcinoma. Although this cholangiocarcinoma is caused by both physical and chemical irritation from direct contact with adult worms and their excretory–secretory products (ESPs), the precise molecular events of the host–pathogen interactions remain to be elucidated. To better understand the effect of C. sinensis infection on cholangiocarcinogenesis, we profiled the kinetics of changes in cancer-related microRNAs (miRNAs) in human cholangiocarcinoma cells (HuCCT1) treated with C. sinensis ESPs for different periods. Using miRNA microarray chips containing 135 cancer-related miRNAs, we identified 16 miRNAs showing differentially altered expression following ESP exposure. Of these miRNAs, 13 were upregulated and 3 were downregulated in a time-dependent manner compared with untreated controls. Functional clustering of these dysregulated miRNAs revealed involvement in cell proliferation, inflammation, oncogene activation/suppression, migration/invasion/metastasis, and DNA methylation. In particular, decreased expression of let-7i, a tumor suppressor miRNA, was found to be associated with the ESP-induced upregulation of TLR4 mRNA and protein, which contribute to host immune responses against liver fluke infection. Further real-time quantitative PCR analysis using ESP-treated normal cholangiocytes (H69) revealed that the expressions of nine miRNAs (miR-16-2, miR-93, miR-95, miR-153, miR-195, miR-199-3P, let7a, let7i, and miR-124a) were similarly regulated, indicating that the cell proliferation and inhibition of tumor suppression mediated by these miRNAs is common to both cancerous and non-cancerous cells. These findings constitute further our understanding of the multiple cholangiocarcinogenic pathways triggered by liver fluke infection.
KeywordsClonorchis sinensis Cholangiocarcinoma MicroRNA array Excretory–secretory products
We thank Drs. Dae Ghon Kim for donating H69 cells and Hye Ryung Jun for assistance in the early phases of the study. This study was supported by a National Research Foundation of Korea (NRF) grant funded by the Korea Government (MEST) (No. 2012R1A2A2A01014237).
Conflict of interest
The authors declare that they have no conflict of interest.
- Braconi C, Huang N, Patel T (2010) MicroRNA-dependent regulation of DNA methytransferase-1 and tumor suppressor gene expression by interlukin-6 in human malignant cholangiocyte. Hepatol 51:881–890Google Scholar
- Du WW, Fang L, Li M, Yang X, Liang Y, Peng C, Qian W, O’Malley YQ, Askeland RW, Sugg SL, Qian J, Lin J, Jiang Z, Yee AJ, Sefton M, Deng Z, Shan SW, Wang CH, Yang BB (2013) MicroRNA miR-24 enhances tumor invasion and metastasis by targeting PTPN9 and PTORF to promote EGF signaling. J Cell Sci 126:1440–1453PubMedCrossRefGoogle Scholar
- Esquela-Kerscher A, Slack FJ (2006) Oncomirs—microRNAs with a role in cancer. Nature Rev 6:259–269Google Scholar
- Karakatsanis A, Papaconstantinou I, Gazouli M, Lyberopoulou A, Polymenneas G, Voros D (2013) Expression of microRNAs, miR-21, miR-31, miR-122, miR-145, miR-146a, miR-200c, miR-221, miR-222, and mir-223 in patients with hepatocellular carcinoma or intrahepatic cholangiocarcinoma and its prognostic significance. Mol Carcinog 52:297–303PubMedCrossRefGoogle Scholar
- Kim DW, Kim JY, Moon JH, Kim KB, Kim TS, Hong SJ, Cheon YP, Pak JH, Seo SB (2010) Transcriptional induction of minichromosome maintenance protein 7 (Mcm7) in human cholangiocarcinoma cells treated with Clonorchis sinensis excretory-secretory products. Mol Biochem Parasitol 173:10–16PubMedCrossRefGoogle Scholar
- Liu H, Brannon AR, Reddy AR, Alexe G, Seiler MW, Arreola A, Oza JH, Yao M, Juan D, Liou LS, Ganesan S, Levine AJ, Rathmell WK, Bhanot GV (2010a) Identifying mRNA targets of microRNA dysregulated in cancer: with application to clear cell renal cell carcinoma. BMC Syst Biol 4:51PubMedCentralPubMedCrossRefGoogle Scholar
- Liu X, Sempere LF, Ouyang H, Memoli VA, Andrew AS, Luo Y, Demidenko E, Korc M, Shi W, Preis M, Dragnev KH, Li H, DiRenzo J, Bak M, Freemantle SJ, Kauppinen S, Dmitrovsky E (2010b) MicroRNA-31 functions as an oncogenic miRNA in mouse and human lung cancer cells by repressing specific tumor suppressors. J Clin Invest 120:1298–1309PubMedCentralPubMedCrossRefGoogle Scholar
- Wei Y, Nazari-Jahantigh M, Chan L, Zhu M, Heyll K, Corbalán-Campos J, Hartmann P, Thiemann A, Weber C, Schober A (2013) The microRNA-342-5p fosters inflammatory macrophage activation through an Akt1- and microRNA-155-dependent pathway during atherosclerosis. Circulation 127:1609–1619PubMedCrossRefGoogle Scholar