| 摘要: | 本研究主要目是探討DEPTOR (DEP domain containing mTOR - interacting protein)與乳癌的關聯性。先前有研究利用microarray的方式觀察基因在各種癌症組織和腫瘤細胞中的表達情形，其中發現DEPTOR大量表現在乳癌細胞中。但目前對於DEPTOR與乳癌致癌的相關性研究很少。因此，本論文的主旨是探討DEPTOR在乳癌中扮演的角色與影響。初步利用RT-PCR和Real-Time PCR的方式在13株乳癌和正常細胞株與140名乳癌病患檢體，發現DEPTPR會大量表達在腫瘤細胞株與病患的腫瘤檢體。從臨床分析結果發現DEPTOR與ER陽性有相關並由免疫組織染色觀察發現DEPTOR大量表現在腫瘤組織中。接著處理Estrogen發現DEPTOR的表達顯著增加，進一步利用promoter assay和ChIP assay的方式證實ERα、SP1和p300加入Estrogen後調控DEPTOR的表現。使用siRNA的方式knockdown 降低DEPTOR的表達抑制乳癌細胞的生長。進一步在穩定表達si-DEPTOR的細胞處理Tamoxifen藉由西方墨點法和MTT assay證實DEPTOR在抗癌藥物的敏感度上可能扮演重要的角色。最後找出可以抑制DEPTOR表達的天然萃取物進而改善乳癌病患的治療或降低腫瘤復發。總言之，本實驗的結果更了解此基因在乳癌的重要性，對未來治療乳癌病患可以有多一些的貢獻。
The main purpose of this study is to explore the DEPTOR (DEP domain containing of mTOR - interacting protein) associated with breast cancer. Previous studies using microarray way to view large number of each gene expression in cancer tissues and cells, found to DEPTOR overexpression in breast cancer cells. But, DEPTOR research few in breast cancer. Therefore, the main thrust of this paper is to explore the DEPTOR play a role in breast cancer and impact. Therefore, the main thrust of this paper is to explore DEPTOR play a role in breast cancer and influence. In 13 breast cancer and normal cell lines and 140 breast cancer patient samples by RT-PCR and Real-Time PCR, discovery DEPTPR high expression in tumor cell lines and patient tumor samples. Results from clinical analysis DEPTOR with ER-positive related by immunohistochemical staining was found DEPTOR overexpression in tumor tissue. Then treatment estrogen significant increase found DEPTOR expression, further use of promoter assay and ChIP assay confirmed promoter activity of ERα, SP1 and p300 treatment estrogen regulation DEPTOR. SiRNA knockdown reduce DEPTOR expression to inhibit breast cancer cell growth. Further expression si-DEPTOR's in a stable cell treatment Tamoxifen confirmed by western blotting and MTT assay DEPTOR may play an important role in the anti-cancer drug sensitivity Finally, identify natural extracts can inhibit DEPTOR expression thus improving treatment of breast cancer patients or reduce tumor recurrence. In summary, the results of this experiment a better understanding of importance of this gene in breast cancer, can have some contribution to future treatment of breast cancer patients. |
