| Summary: | 碩士 === 中國醫藥大學 === 癌症生物學研究所碩士班 === 100 === A Disintegrin And Metalloprotease (ADAM) 9 has been implicated in cancer progression of various solid tumors, and this phenomenon also discovered in the renal cell carcinoma (RCC). Hence, we would like to explore the role of ADAM9 in the enhancement of malignant renal cancer cells transition. We noticed no difference in cancer cell proliferation but alter the migration and invasion abilities of renal cancer cell carcinoma after knockdown of ADAM9 expressions. Interestingly, RCC42shADAM9 revealed enhancement of cell migration but reduction of invasion activities. In contrast, RCC52shADAM9 demonstrated the decrease migration with no alteration in the invasion activities when compared to RCC52shGFP. These opposite cell behaviors of RCC42 and RCC52 in response to ADAM9 ablation indicated different ADAM9 regulatory molecules involved in the cell behavior changes. To investigate the role of ADAM9 expression in both cells, we first determine the ADAM9 and integrins in regulation of cell adhesion on collagen. The result demonstrated similar enhancement of cell adhesion activities in both RCC42shADAM9 and RCC52shADAM9 compared
to control cell lines. Nor did we found the alteration of integrin composition after knockdown of ADAM9 expression in both cell lines, suggesting the regulation of RCC42 and RCC52 activities is not through the regulation of ectodomain of ADAM9. Furthermore, it has been demonstrated the interaction of PKC-δ, MAD2β, SNX9 or endophilin-1 with cytoplasmic domain of ADAM9. To evaluate these interactions in RCC42 and RCC52, gene expression analyses at transcription and translation were conducted and revealed similar expression of MAD2β, SNX9 in both RCC cell lines. However lost of endophilin-1 expression in RCC42 was detected in both qPCR and western blot studies, suggesting the interaction of ADAM9 and endophilin-1 might
regulate cell migration and invasion activities in RCCs. We found that endophilin-1 were enhanced RCC42 cell migration and invasion ability once we overexpress endophilin-1 in RCC42. However it does not affect cell proliferation. Immunofluorescence analyses of ADAM9 and endophilin-1 demonstrated that ADAM9 does not co-location with endophilin-1. In conclusion, we proved that ADAM and endophilin-1 regulate cell migration and invasion through an independent pathway.
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