| Summary: | 碩士 === 國立中興大學 === 生命科學系所 === 94 === Recently, the most severe disease, cancer, has attacked human and cause lots of people dead in every years and getting worse. A lot of pharmacologists try to find the answers from the traditional Chinese herb medicines. However, there is no specific and high-throughput way to screen thousands of these “health-care” herbs to unknown anti-cancer pathway. According to our previous microarray’s studies, we have identified an oncosuppressor gene (HLJ1) which was reduced expression in highly metastatic lung cancer cells and might relate to patients’ survival rate. In this study, the selected herbs were screened for their role in regulating the promoter activity of the HLJ1 and IL-8. First, we used MTT assay to determine cytotoxicity of ninety traditional Chinese medicines in CL1-0 cells. Subsequently, we transfected HLJ1 and IL-8 promoter construct to CL1-0 cells and then treated with the IC20 concentration of the herb medicines. After 24 hours, we measured the luciferase activity by reporter gene assay. To confirm the results of reporter assay, we carried out the western blotting of HLJ1 in CL1-0 and CL1-5 cells. Then we further performed wound-healing assay (migration assay) and invasion assay to investigate which traditional Chinese medicines can suppress cancer cell metastasis. We have identified thirty-six traditional Chinese herb medicines which can enhance HLJ1 promoter activity in CL1-0 and twenty-five traditional Chinese medicines which can enhance HLJ1 promoter activity in CL1-5. Furthermore, thirty-seven herb medicines could induce HLJ1 protein expression in CL1-0 or CL1-5. We also found that there are five of the thirty-seven Chinese herb medicines can suppress cancer cell migration and invasion. These Chinese herb medicines may induce HLJ1 promoter activity through affecting YY-1 and AP-1 protein binding affinity. In addition, our results also revealed that there are fifteen Chinese herb medicines that can suppress IL-8 promoter activity. Electromobility shift assay further confirmed that there are three Chinese herb medicines may suppress IL-8 promoter activity through inhibiting NF-κB and AP-1 binding affinity.
In this study, we have set up a HLJ1 and IL-8 targeting drug-screening platform respectingly, which could be used to screen thousands of traditional Chinese herb (heath-care) medicines. Our results showed that there are several herb medicines that can up-regulate HLJ1 and down-regulate IL-8 promoter activity and suggested that these herb medicines may have the important elements that can indirectly or directly induce HLJ1 or repress IL-8 expression to suppress cancer cell migration and invasion.
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