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The role of circ-WHSC1-mediated classical competing endogenous pathway and non-classical pathway in the metastasis of colorectal cancer.
China (IFMSA-China) - Soochow University, Suzhou
Department of Pathology and Pathophysiology
Type of Research Project
- Basic science
What is the background of the project?
Metastasis is the main cause of death in patients with colorectal cancer (CRC). Recently, we identified a circular Ribonulceic acid (RNA), circ-NSD2 (nuclear receptor binding SET domain protein 2) (aka. circ-WHSC1), by mouse model of liver metastasis of CRC and RNA (transcriptome)-sequencing, which was significantly elevated in the liver metastasis than in situ tumors of CRC. In vivo and in vitro experiments showed that circ-NSD2 contributes to the metastasis of CRC. Interestingly, circ-NSD2 plays not only as microRNA (miRNA) sponge for miR-199b-5p via a classical competing endogenous RNA pathway but also as a key regulator of H3K36me2. Based on these findings, we propose a key functional role of circ-NSD2 in CRC metastasis by a classical competing endogenous pathway and a non-classical pathway.
What is the aim of the project?
Our study will uncover circ-NSD2 as a new functional molecule in metastasis of CRC by mediating a classical competing endogenous pathway and a non-classical pathway.
What techniques and methods are used?
1. Cell culture: we will use several cell models in the program, including SW480, LoVo, Rko; 2. To determine the expression of gene expression, we will use: λ Realtime Polymerase Chain Reaction (PCR): to test the mRNA levels of the target genes; λ Western blot: to test the protein levels of the target genes; λ Luciferase Reporter assay: used as a tool to study gene expression at the transcriptional level; λ ChIP (Chromatin Immunoprecipitation) experiment: used to investigate the interaction between proteins and DNA; 3. To observe metastatic features of CRC cells, we will use: λ Transwell assay: to test the migration/invasion ability of cancer cells; λ Animal experiments: using nude mice to test the metastasis of cancer cells; HE/IHC HE (hematoxylin-eosin staining) / IHC (Immunohistochemistry): testing the pathological features of the clinical tissue samples.
What is the role of the student?
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
The student will: 1. Learn how to build a scientific hypothesis and then how to test it; 2. Learn the relevant steps of the cell culture and be able to conduct cell culture independently. 3. Learn basic biochemistry and molecular biology techniques including cell culture techniques, real-time PCR, Western Blot, Transwell assay, HE/IHC, Reporter Assay, ChIP; 4. Learn animal experiment techniques; 5. Analyze collected data with IBM SPSS® (Statistical Package for Social Sciences) and interpretation methods. Conduct literature retrieval and reading, and learn to use software including Endnote, ImageJ, SPSS, and GraphPad Prism.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
There will be preliminary readings provided for the student before arrival; The graduate students at the lab will be responsible for most of the teaching work of the experiment; There will be a group meeting every other week, held by the tutor.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a scientific report
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
No specific skills required; Subjects passed: Molecular Biology ; Previous experience with: Lab research
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months) - Pre-Medical students from the American-British system - Students in biomedical fields
- Chen LY; Zhi Z; Wang L; Zhao YY; Deng M; Liu YH; Qin Y; Tian MM; Liu Y; Shen T; Sun LN*; Li JM*. NSD2 circular RNA promotes metastasis of colorectal cancer by targeting miR-199b-5p mediated DDR1 and JAG1 signaling. J Pathol. 2019 May;248(1):103-115. doi: 10.1002/path.5238.
- Sun LN#; Zhi Z#; Chen LY; Zhou Q; Li XM; Gan WJ; Chen S; Yang M; Liu Y; Shen T; Xu Y; Li JM. SIRT1 suppresses colorectal cancer metastasis by transcriptional repression of miR-15b-5p. Cancer Lett. 2017 Nov 28; 409:104-115.
- Sun LN#; Xing C#; Zhi Z; Liu Y; Chen LY; Shen T; Zhou Q; Liu YH; Gan WJ; Wang JR; Xu Y; Li JM. Dicer suppresses cytoskeleton remodeling and tumorigenesis of colorectal epithelium by miR-324-5p mediated suppression of HMGXB3 and WASF-2. Oncotarget. 2017 May 25; 8(34):55776-55789.
- Sun L#; Fan Z#; Chen J#; Tian W; Li M; Xu H; Wu X; Fang M; Xia J; Xu Y. Protein inhibitor of activated STAT 4 (PIAS4) regulates pro-inflammatory transcription in hepatocytes by repressing SIRT1. Oncotarget. 2016 Jul 12; 7(28):42892-42903.
- Sun L#; Fan Z#; Chen J#; Tian W; Li M; Xu H; Wu X; Shao J; Bian Y; Fang M; Xu Y. Transcriptional repression of SIRT1 by protein inhibitor of activated STAT 4 (PIAS4) in hepatic stellate cells contributes to liver fibrosis. Sci Rep. 2016 Jun 21; 6:28432.
- Sun L; Li H; Chen J; Dehennaut V; Zhao Y; Yang Y; Iwasaki Y; Kahn-Perles B; Leprince D; Chen Q; Shen A; Xu Y. A SUMOylation-dependent pathway regulates SIRT1 transcription and lung cancer metastasis. J Natl Cancer Inst. 2013 Jun 19;105(12):887-98.
- Sun L#; Li H#; Chen J#; Iwasaki Y; Kubota T; Matsuoka M; Shen A; Chen Q; Xu Y. PIASy mediates hypoxia-induced SIRT1 transcriptional repression and epithelial- to- mesenchymal transition in ovarian cancer cells. J Cell Sci. 2013 Sep 1; 126(Pt17): 3939-47.
- Zhang X; Chen J; Sun L*; Xu Y*. SIRT1 deacetylates KLF4 to activate Claudin-5 transcription in ovarian cancer cells. J Cell Biochem. 2018 Feb; 119(2):2418-2426.
- Liu L; Chen J; Sun L*; Xu Y. RhoJ promotes hypoxia induced endothelial-to- mesenchymal transition by activating WDR5 expression. J Cell Biochem. 2018 Apr; 119(4):3384-3393.
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