| Summary: | 碩士 === 嘉南藥理科技大學 === 生物科技系暨研究所 === 95 === Lung cancer is most common cause of cancer death in Taiwan. Human Liver DnaJ-like protein (HLJ1) is a tumor suppressor gene in non–small-cell lung cancer (NSCLC) and its highly expression of HLJ1 is associated with reduced cancer recurrence and prolonged survival of NSCLC patients. Recent studies have indicated that HLJ1 expressed in NSCLC cells would inhibit cell proliferation, anchorage-independent growth, cell motility, invasion, and tumorigenesis. The HLJ1 DNA sequence analysis showed that it contained the J and G/F domain sequence of the Heat-shock protein 40 (Hsp40) family members. The amino-acid sequencing revealed that HLJ1 might be a member of heat shock protein family. Yet it has not been investigated that how heat shock factor can regulate HLJ1 expression in response to many stresses, such as heat shock, serum deprivation, cell density, exposure to heavy metals, sodium arsenite, and oxidative stress. Then we observed that the expression of HLJ1 in CL1-0 and CL1-5 adenocarcinoma cells was responsive to heat shock stress, cell-density, serum deprivation, pH-mediated stimuli and glutamine treatment before heat shock stress. For heat shock, serum deprivation, glutamine treatment experiment, CL1-0 and CL1-5 cells were plated on different dish in different count of cell number, and incubated at diverse conditions. In order to determining the critical region in HLJ1 promoter involved in heat shock response, the promoter region of HLJ1 was cloned into pGL3-basic vector. The Luciferase reporter assay was performed to detect the promoter activity of HLJ1. Furthermore, we will analyze the putative heat shock factor element (HSE) of HLJ1 promoter and construct various deletions of HLJ1 promoter. These studies imply stresses such as heat shock, high cell density, serum deprivation, pH-mediated and glutamine cultures are important factors involved in the up-regulation of HLJ1 expression.
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