乳癌專一性單株抗體的製備與功能之探討

Generation and Characterization of Monoclonal Antibodies against Breast Cancer

戴慈文(Tzu-Wen Tai)

指導教授 ： 吳漢忠 ； 林欽塘

國立臺灣大學/醫學院/病理學研究所/碩士(2008年)

https://doi.org/10.6342/NTU.2008.10441

摘要

乳癌為先進國家中女性常見的癌症。在美國，乳癌排名女性癌症罹患率的第一位，同時也是女性癌症死亡率第二位。而在台灣，乳癌為女性癌症死因之第四名。乳癌同時也是造成女性死亡年齡提早的主要癌症死因之一。目前乳癌在早期治療後，五年存活率可高達90%以上，然而發生轉移後的五年存活率則降至50%以下。乳癌常見的治療方式有外科手術切除、放射線治療以及化學治療。手術切除與放射線治療較適用於早期治療，待腫瘤發生轉移或再度復發後，化學治療為較常見的治療方式。然而化學治療常使病人產生較大的副作用，因此療效有限。近年來，由於治療性抗體相關的研究與發展，發現治療性抗體能更專一性地針對腫瘤細胞並使癌症治療能更有效進行。目前美國食品藥物管理局已認可多種治療性抗體，並使之得以使用於臨床上治療癌症。在本篇研究中，我們建立 11種單株抗體可專一性辨認乳癌細胞 MDA-MB-231，並對正常細胞具極低的親和性。我們也經由不同的方法分析並探討這些單株抗體之特性。在對11種不同癌細胞株的酵素免疫分析法 (ELISA)實驗中，測得單株抗體 MDA-Ab 1-1, MDA-Ab 2-6, MDA-Ab 3-1, MDA-Ab 5-9, MDA-Ab 6-9, MDA-Ab 7-2, MDA-Ab 8-1,MDA-Ab 9-2, MDA-Ab 10-3 及 MDA-Ab 11-1 表現出不同目標蛋白質結合反應。我們再藉由 ELISA、西方墨漬法、免疫染色及流式細胞測定，得知這些單株抗體之特性。經由西方墨漬法的實驗結果，這些單株抗體辨認出不同分子量的標的蛋白。而根據細胞與組織切片染色，我們清楚確認有些單株抗體的標的蛋白表現在腫瘤細胞的細胞膜上。這些單株抗體對腫瘤具有高度的專一性，因此有潛力在未來應用於診斷乳癌病人及發展標靶治療。

關鍵字

乳癌 ； 單株抗體 ； 治療性抗體 ； 標靶治療

並列摘要

Breast cancer is the most commonly diagnosed cancer in women and the second leading cause of cancer deaths in American women. In Taiwan, breast cancer is higher than those of other cancers in women aged 40 to 60 years and is the fourth leading cause of cancer deaths. The five-year survival rate of localized breast cancer cases is higher than 90% while that of the metastatic status will be below 50%. The breast carcinoma in situ is usually treated by surgical mastectomy and lumpectomy combined with radiotherapy. The major treatment for metastatic breast cancer is chemotherapy nowadays. However, the traditional chemotherapy may be accompanied with several side effects, including nausea, vomiting, diarrhea, hair loss, weight loss, fatigue, anemia, edema, heart failure and liver and kidney damage. Furthermore, chemotherapy can easily induce chemoresistent outcomes in cancer cells. Based on the disadvantages of chemotherapy, it is necessary to develop new modalities of therapy such as targeted therapies.
During the past decades, therapeutic antibodies have been applied extensively in clinical treatment of cancers due to the well-developed technology of generating monoclonal antibodies (mAbs). Up to now, several mAbs have been approved by the US Food and Drug Administration (FDA) for used in various clinical settings, including cancer therapy. In this study, we have generated 11 mAbs (MDA-Ab 1-1, MDA-Ab 2-6, MDA-Ab 3-1, MDA-Ab 5-9, MDA-Ab 6-9, MDA-Ab 7-2, MDA-Ab 8-1,MDA-Ab 9-2, MDA-Ab 10-3 and MDA-Ab 11-1) which exhibited higher binding affinities for cancer cells than those for normal cells. The specificity of these mAbs was subsequently confirmed by the ELISA, Western blotting, immunostaining and flow cytometric assays. The results from Western blot analysis revealed that these mAbs recognized different target proteins. The localization of these target proteins on the plasma membrane of cancer cells were further confirmed using cancer cell lines and human surgical specimens of breast cancer. In conclusion, we have generated several mAbs against breast cancer cells. These specific mAbs are useful to identify the tumor antigens or develop ligand-targeted therapeutics for breast cancer.

並列關鍵字

breast cancer ； monoclonal antibody ； therapeutic antibody ； ligand-targeted therapy

參考文獻

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