微流體晶片應用於個人化乳癌化療藥物敏感性測定之研究

Individualized Chemosensitivity Profiling of Breast Cancer Tissue Using a Microfluidic Assay

王成禾(Chen-Ho Wang)

指導教授 ： 胡文聰

國立臺灣大學/工學院/應用力學研究所/碩士(2011年)

https://doi.org/10.6342/NTU.2011.03066

摘要

化學治療是癌症的最主要治療方式之一，化療藥物的種類繁多，特定病患最適合的化療藥物選用在進行化學治療前尤其重要，若施予不適合病患的化療藥物，則會導致不佳的治療效果，使病患必須進行多次的化學治療，造成病患身體上的沉重負擔。
基於上述之原因，本研究發展一種可進行化學治療藥物測試實驗的微流體晶片，進行癌症病患檢體的化學藥物測試 (Chemosensitivity Microfluidic Assay, CheMA)。期望藉由自癌症病患病灶取出的檢體，在化學治療前進行個別病患的最適化療藥物種類與劑量的預測。本研究著重於有效利用自癌症病患取出的少量檢體，做最多有效的化學治療藥物測試，期望得以提升每一次化學治療的效果，實現有效地在手術前縮減腫瘤體積及手術後清除剩餘癌細胞。
檢體自病灶位置取出後，便經由處理製成配於培養液的細胞懸浮液並種入本研究中發展的微流體晶片，並施以兩種主要使用的乳癌化學治療藥物 (cisplatin, docetaxel) 進行藥物反應測試。藉由微流體技術，本藥物測試方法共僅需大約9,000顆細胞即可完成一次單一藥物化療藥物試驗。除了所需的細胞數目較低之外，為使植入的少量細胞可以在進行液體置換時有效的存留在晶片上，本元件設計一環形圍繞細胞植入區的微流道。藉由於微流道進行液體置換，可避免換液體所使用的工具直接碰觸到細胞，以此避免明顯的細胞數目改變，造成藥物測試結果的不準確。細胞被施以化學治療藥物四十八小時後，螢光染劑即置入以判定細胞在不同藥物濃度處理下的存活率。MCF7及MDA-MB-231細胞株的藥物反應測試實驗，在本研究中用以驗證本微流體晶片作為藥物測試平台的可行性。
根據細胞株的藥物測試實驗結果，在CheMA上進行之藥物測試實驗結果與96孔盤控制實驗結果之間，於統計上沒有顯著差異。此外，細胞株於兩種化療藥物處理下的IC50數值在兩平台上也相當接近。三個病患檢體藥物反應實驗也在本研究中使用CheMA進行。
簡言之，本微流體晶片及藥物敏感度檢測方法經由細胞株實驗後，在細胞數量極少的情況下進行藥物測試實驗的可行性即被驗證。藉由本研究所發展的微流體晶片及實驗方法，期望可以應用於病人檢體的藥物測試，以提供臨床醫師在藥物選用上的有效建議。

關鍵字

微流體 ； 化療敏感性檢測 ； 個人化化學治療 ； 藥物反應檢測 ； 初代細胞培養

並列摘要

There are many different kinds of anticancer drugs for chemotherapy nowadays. The most important issue to doctors is the choice of anticancer drug for specific patients. Once patients were treated with ineffective treatment, the patients would need further repeats of chemotherapy which was really harmful to patients.
In order to solve the problem stated above, we present a microfluidic device to conduct chemosensitivity microfluidic assay (CheMA) for primary cell-based individualized chemosensitivity profiling while consuming 9,000 cells for single reagent response assay. Cancer cells from patients were seeded into microfluidic device and treated with 3 concentrations of 2 major used anticancer drugs. Roughly, only 9,000 cells in total were required for a single drug response assay by CheMA. In addition, cells in units of microfluidic device were conserved by conducting solution exchange through surrounding microchannels. After drug treatment, cell viability was quantified by a tri-staining method to distinguish cancer cells, normal cells and dead cells. MCF7 and MDA-MB-231 cell lines were applied to anticancer drug screening to prove the feasibility of drug response assay by CheMA.
No statistical significance was found in dose-responses between CheMA and 96-well plate controls. Furthermore, the IC50 values of cisplatin and docetaxel were similar between the two platforms. Moreover, the drug response of patient samples were conducted by CheMA with few cells demanded. Primary samples of breast cancer patients were also examined by CheMA.
In short, CheMA could be applied to prediction of individual chemotherapy within days when the sizes of tumors were not enough for a complete traditional assay.

並列關鍵字

Microfluidics ； Chemosensitivity profiling ； Individualized chemotherapy ； Drug response assay ； Primary cell culture

參考文獻

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