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| ???org.dspace.app.webui.jsptag.ItemTag.dcfield??? | Value | Language |
|---|---|---|
| dc.contributor.advisor | 許麗卿 | |
| dc.contributor.author | Ying-Hsi Lin | en |
| dc.contributor.author | 林映希 | zh_TW |
| dc.date.accessioned | 2021-06-08T07:08:02Z | - |
| dc.date.copyright | 2011-10-07 | |
| dc.date.issued | 2011 | |
| dc.date.submitted | 2011-08-12 | |
| dc.identifier.citation | Agarwal, R., and Kaye, S.B. (2003). Ovarian cancer: strategies for overcoming resistance to chemotherapy. Nat Rev Cancer 3, 502-516.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/26374 | - |
| dc.description.abstract | 分子標靶藥物結合傳統細胞毒殺治療的應用在增強抗癌反應上似乎已成為新的治療策略。於人類的癌症中常見到PI3K/Akt/mTOR訊息途徑失常，且此訊息途徑異常也容易導致對化學治療產生抗藥性。MK-2206為一Akt抑制劑，可抑制癌細胞的增生。
於此，我們證實了MK-2206與cisplatin或taxol的合併治療可提升cisplatin或 taxol對卵巢癌細胞SKOV3 (Akt表現活化)和ES2 (Akt未表現活化)的作用。於taxol或cisplatin的治療中再給予MK-2206可加強抑制細胞增生的作用。合併治療可能是藉由抑制Akt及其下游受質4E-BP1和p70S6K的磷酸化，恢復因taxol或cisplatin所降低的p53表現，或減低Bcl-2蛋白來提升藥效。此外，我們於兩株卵巢癌細胞中都發現MK-2206可提高細胞內活性氧化物的生成，這可能會導致細胞凋零。另一方面，MK-2206可抑制BRCA1在cisplatin所造成的DNA損壞處聚集，這可能會使DNA無法修復。 所有的結果指出，Akt抑制劑MK-2206可能透過不同的機制來影響不同的卵巢癌細胞。不論癌細胞的Akt活化與否，MK-2206都可提升現在常用於臨床上之細胞毒殺劑之效用。 | zh_TW |
| dc.description.abstract | The application of molecular targeted agents in combination with traditional cytotoxic regimens seems to be a promising therapeutic strategy to increase antitumor response. Abnormalities in the PI3K/Akt/mTOR signaling pathway are common in human cancers and contribute to chemotherapy resistance. MK-2206 is an Akt inhibitor known to inhibit cancer cell proliferation.
Here, we demonstrate an enhanced response to combination treatment of MK-2206 with either cisplatin or taxol in the ovarian cancer cell lines SKOV3 (with constitutively active Akt) and ES2 (with inactive Akt). Taxol or cisplatin followed by MK-2206 treatment synergistically inhibited cell proliferation. This combination was effective in promoting cell death by inhibiting Akt phosphorylation and downregulating its downstream effectors, p-4E-BP1 (Thr37/46) and p-p70S6K (Thr389), in SKOV3 cells, restoring p53 level which was downregulated after taxol or cisplatin treatment in ES2 cells, and downregulating pro-survival Bcl-2 protein in both SKOV3 cells and ES2 cells. In addition, we found that intracellular ROS generation was increased in both ovarian cancer cell lines after MK-2206 treatment, that may lead to apoptosis. Moreover, MK-2206 could suppress BRCA1 foci formation at DNA damage sites induced by cisplatin that may affect DNA repair. The overall results indicate that Akt inhibitor MK-2206 may augment the efficacy of existing cytotoxic agents through different mechanisms in either Akt active or inactive ovarian cancer cells. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T07:08:02Z (GMT). No. of bitstreams: 1 ntu-100-R98423012-1.pdf: 2971063 bytes, checksum: e5f310325d3991e81fdd1b2a7f9a97cd (MD5) Previous issue date: 2011 | en |
| dc.description.tableofcontents | 口試委員會審定書………………………………………………………………… i
致謝………………………………………………………………………………… ii中文摘要…………………………………………………………………………… iii Abstract…………………………………………………………………………….. iv Contents…………………………………………………………………………….. vi List of Figures……………………………………………………………………… viii List of Tables……………………………………………………………………….. x Chapter 1 Introduction………………………………………………………….. 1 1.1 Ovarian cancer and standard treatment……………………………………. 1 1.2 Activation of PI3K/Akt signaling pathway in human cancer and drug resistance…………………………………………………………..…. 3 1.3 MK-2206, an allosteric Akt inhibitor…………………………………….... 7 1.4 BRCA1 and DNA repair............................................................................... 8 Chapter 2 Objectives…………………………………………….......................... 20 Chapter 3 Materials and methods……………………………………………..…. 23 3.1 Drugs and chemicals……………………………………………………..…. 23 3.2 Cell lines and cell culture…………………………………………………… 23 3.3 Cell growth and MTT cytotoxicity assay………………………………...… 23 3.4 Flow cytometric analysis of cell cycle and apoptosis……………………… 24 3.5 Western blot analysis……………………………………………………….. 25 3.6 Detection of intracellular reactive oxygen species (ROS) generation……… 26 3.7 Immunofluorescence staining………………………………………………. 26 3.8 Combination index (CI) analysis…………………………………………… 27 3.9 Statistical analysis………………………………………………………….. 27 Chapter 4 Results…………………………………………………………………. 30 4.1 MK-2206 potentiates taxol induced growth inhibition in a drug sequence dependent manner………………………………………………… 30 4.2 p-Akt and its downstream effectors are downregulated by taxol and MK-2206 combination in Akt constitutively active cells……………... 31 4.3 The pro-survival Bcl-2 protein level is diminished by taxol/MK-2206 treatment in SKOV3 and ES2 cells and the p53 protein level downregulated by taxol is reversed by taxol/MK-2206 combination in ES2 cells………..… 32 4.4 ROS is a mediater of the MK-2206 stimulated apoptosis………………….. 32 4.5 MK-2206 potentiates cisplatin induced growth inhibition in a drug sequence dependent manner………………………………………………… 33 4.6 p-Akt and its downstream effectors are downregulated by cisplatin and MK-2206 combination in Akt constitutively active cells…….. 34 4.7 The pro-survival Bcl-2 protein level is diminished by cisplatin/ MK-2206 treatment in SKOV3 and ES2 cells and p53 protein level downregulated by cisplatin is reversed by cisplatin/ MK-2206 combination in ES2 cells…………………………………..…….. 35 4.8 ROS is a mediater of the MK-2206 stimulated apoptosis………………….. 36 4.9 MK-2206 disrupts BRCA1 foci formation at DNA double-strand breaks induced by cisplatin……..…………………………………………... 36 Chapter 5 Discussion……………………………………………………………… 54 Chapter 6 Conclusion……………………………………………………………... 58 References…………………………………………………………………………….. 60 | |
| dc.language.iso | en | |
| dc.title | Akt抑制劑MK-2206可提升taxol和cisplatin對卵巢癌細胞的毒殺性 | zh_TW |
| dc.title | An Akt inhibitor MK-2206 enhances the cytotoxicity of taxol and cisplatin in ovarian cancer cells | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 99-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 陳燕惠,沈麗娟 | |
| dc.subject.keyword | Akt抑制劑,卵巢癌, | zh_TW |
| dc.subject.keyword | Akt inhibitor,ovarian cancer, | en |
| dc.relation.page | 66 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2011-08-12 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 藥學研究所 | zh_TW |
| Appears in Collections: | 藥學系 | |
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