Summary: | 碩士 === 國立臺北科技大學 === 化學工程與生物科技系生化與生醫工程碩士班 === 104 === Androgen-receptor splice variant 7 (AR-V7) messenger RNA, which are highly associated with resistance to hormone target drugs (enzalutamide and abiraterone) in advanced prostate cancer. Therefore, we aim to develop a biosensor that can rapidly discriminate resistant statuses of prostate cancer cells. We utilized electrolyte-insulator-semiconductor (EIS) structures feature an high sensitivity Yb2Ti2O7 sensing membrane and used three steps of sensing membrane fixation methods, including: (1) samples were encapsulated on the EIS by agarose gel; (2) sensing membrane surface treated with 3-aminopropyltriethoxysilane, and glutaraldehyde. The 5- amino modified oligonucleotide probe coupled onto the membrane; and (3) each procedure should undergo electric field to align and improve binding efficiency of molecules. The sensing membrane binding status were analyzed by measuring voltage-capacitance (C-V) curves using LCR meter. The results demonstrated that electric field based sensing surface functionalized, established a good stability (95%) detection platform, successfully improved the unstable situations of agarose gel method. The resistant prostate cancer can be identified by differences C-V curves, through cDNA hybridized with complementary or non-complementary AR-V7 probe. Comparison of conventional fluorescent in situ hybridization (FISH), the method in this study, provide a stable, rapid, simple, label-free and cost effect method to detect genetic variation. Rapid identification of resistant statuses in prostate cancer patients is essential for clinicians to decide whether hormone therapy will be the treatment of choice, therefore it provided the high potential to serve as a diagnosis tool in clinical practice in the future.
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