| Huwel is a E3 ubiquitin ligase, containing HECT domain.Ubiquitin ligase is one of three key enzymes necessary for the process of ubiquitination, Which is refered to as the process of specific modification of protein with the help of ubiquitin activating enzyme E1, the desmoenzyme E2,ligase E3 and a series of enzymes, playing great role during protein degradation and function adjustment. Ubiquitin ligase containing HECT domain is considered the important regulatory factor in the process of cancer development. As a E3 ubiquitin ligase containing HECT domain, Huwel has also been regarded as important regulatory factor of cancer, however people share different views on the role Huwel plays in proliferation, apoptosis and cancer development.The protein Huwel encoding is too large, up to 500kd, as a result the full gene cloning kept unfinished for quite a long time until 2005 when two labs cloned the full gene of Huwel.At the same time,they confirmed Huwel as E3 ubiquitin ligase, and found its substrate. But different opinions appeared in the articles published by the two laboratories on the role Huwel plays in proliferation, apoptosis and cancer development.Later researches found that the substrates of Huwel included p53,Mcl-1,N-MYC,C-MYC,histones, CDC6 and so on, which contained both tumor suppressor genes and oncogenes. And in human cancer specimens, the results are controversial. In gastric cancer, prostate cancer, and breast cancer, Huwel expression is higher, implying that Huwel may act as oncogenes.In teratoma cells,Huwel expression is obviously lower, implying that Huwel could be a tumor suppressor gene. Growing studies do not make Huwel’s specific function more clear,on the contrary makes the problem more complicated. So the specific function of Huwel needs further research, and expounding this problem will be of significance to related cancer pathogenesis.As female reproductive system cancer, the malignant and mortality rate of ovarian cancer is high and the rearch of Huwel’s function in ovarian cancer is rare, which makes ovarian cancer the ideal role for research of Huwel’s function in tumorigenesis and cancer development.Researches show that about 90% of ovarian cancer originates in the coverage of ovarian epithelial and the most common genetic variants in ovarian cancer is gene amplification of C-Myc together with K.-RAS and P53 mutations. Due to this, we acquireed virus expressing C-Myc,K-RAS and silencing p53 gene Ihen we injected the virus to Huwel(flop) mouse’s ovaries, attempting to simulate the process of ovarian tumours forming.But Huwel(flop) mouse’s ovarian defects in cleft, leading to virus leakage.So we changed our strategy. We infected Mose cell separated from Huwel(flop) mouse’s ovarian by different viruse, made the Mose cell experience malignant transformation, established Huwel knocked out cell lines noted as Mose-cre (Huwel-) and Huwel contained cell lines Mose-creER (Huwel+).After the establishment of the two cell lines, we compared their morphology, the ability of low serum tolerance, cell migration, colony-forming ability and a series of tumor associated phenotype, trying to analyze the specific role Huwel plays in malignantly transformed MOSE cell. We then tried to analyze the underling mechanism of Huwel’s function. Finally, we speculated the role of Huwel in ovarian tumorigenesis and development. |
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