UTRN与肺癌细胞恶性转化关系的研究生物化学与分子生物学专业论文.docxVIP

哈尔滨工业大学理学硕士学位论文 哈尔滨工业大学理学硕士学位论文 哈尔滨工业大学理学硕士学位论 哈尔滨工业大学理学硕士学位论 PAGE PAGE IV - - - III Abstract In previous study, we demonstrated that Utrophin(UTRN) is a candidate tumor suppressor gene，and the low expression of UTRN occurred in various tumor tissues such as breast cancer, renal cancer and liver cancer, and deletion sometimes occurred. For further studying the expression and location of UTRN protein in cancer cells, we detected the expression of UTRN in lung cancer cell lines by RT-PCR. The results showed that low expression of UTRN cDNA to different extent was observed in 14 samples, and 4 exhibited deletion of UTRN concentrated on cDNA regions from 7909 bp to10277 bp, and 6 demonstrated low expressions in the same region. In additional, the results of Western blotting show that UTRN whose molecular weight is about 130 kD was identified in the nuclei in A549, AGZY-83a and Anip973 cells, while UTRN is high-molecular-weight (395 kD) protein in normal cells. Furthermore, UTRN was detected by immunofluorescence. The results demonstrated that UTRN was translocated to the nuclei in lung cancer cell, while it was mostly distributed at the periphery of the plasma membrane in human embryonic lung cell line MRC-5. As outlined above, the low expression, deletion, nuclear translocation or truncation of UTRN occurred in lung cancer cells. In order to further study the effect of UTRN on the cell proliferation and variation of microfilaments, we silenced the UTRN with RNAi in NIH3T3 cells which was transformed easily. By cell proliferation curve, we found that cell proliferation rate increased significantly, compared to the control. We utilized FITC-phalloidine staining to test microfilaments. The results showed that microfilaments became less and disordered, and the long filopodia was stretched from the cell surface in UTRN knockdown cells. These data are consistent with our previous results of silencing UTRN in HEK293 cells. Above data demonstrate that UTRN plays its f