工學院: 醫學工程學研究所指導教授: 林文澧林憶玲Lin, Yi-LingYi-LingLin2017-03-062018-06-292017-03-062018-06-292016http://ntur.lib.ntu.edu.tw//handle/246246/277533背景：Metformin是一個已經被使用超過半個世紀的口服劑型糖尿病藥物，對病人而言是個便宜又方便的藥物，近十年來，Metformin漸漸被發現有抗腫瘤之效應，越來越多人投入研究其抗腫瘤，甚至是抑制腫瘤幹細胞之機制等；另外，超音波熱效應對腫瘤的抑制效果也廣泛的被研究，超音波同時具有熱效應以及機械效應以改變腫瘤組織的微環境，達到腫瘤抑制的效果。 目的：本研究結合metformin可能殺死癌症幹細胞和治療用超音波改變腫瘤微環境兩種輔助治療，觀察其對小鼠腫瘤治療的效果，期望能夠有效抑制腫瘤的生長。 材料與方法：在細胞實驗，將CT-26細胞放置在96well的培養盤，每個well下入5000個細胞，總共分為四組：Control、Met (1 mM 放置24小時)、pUH (42℃, 0.94 MHz, 0.1 mW and 10%工作週期，治療15分鐘)，和 pUH+Met；在動物實驗中，使用BALB/c公鼠，周齡6至8周，在右背側大腿皮下種CT-26Luc腫瘤細胞株，待腫瘤長至50mm3開始治療，以管餵的方式給予藥物Metformin直到小鼠腫瘤到達1000 mm3，治療用超音波使用1MHz，工作週期50%，強度2.7瓦施打15分鐘，每周兩次，共四次，並觀察紀錄之，隨後將腫瘤取下做組織切片染色。 結果：在細胞實驗中，Met+pUH的組別能夠將CT-26腫瘤細胞的細胞存活率降低至60%，和其他三組在統計上均達到顯著性的差異，動物實驗中的結果顯示，控制組以及管餵Metformin藥物的組別，腫瘤均穩定的生長，有使用治療用超音波治療的組別，則可以在短時間內見到腫瘤的抑制效果，結合Metformin與治療用超音波和其他組相比，均可在腫瘤大小抑制上達到統計上顯著性差異；除此之外，在腫瘤組織的切片上，可以看到各組皆有淋巴球浸潤，但是在有治療用超音波的組別，有更嚴重的組織碎化情形。 總結：不論在細胞實驗或是動物實驗中 ，metformin結合治療用超音波對於腫瘤細胞殺傷力，亦或是小鼠腫瘤大小的抑制，均和其他組別達到統計上的顯著性差異，除了在巨觀的觀察上發現差異之外，在微觀的觀察下，利用顯微鏡觀察各組不同的腫瘤組織切片情形，雖然各組均有淋巴球浸潤之情形，但是超音波治療組細胞排列較為鬆散，且呈螺旋狀分布。 關鍵字：Metformin、超音波熱效應、超音波機械效應、癌腫瘤抑制Background：Metformin is the most widely prescribed drug worldwide for the treatment of type 2 diabetes. Recently, researchers have found that metformin has anti-cancer properties and it selectively targets cancer stem cells. In addition, therapeutic ultrasound has been extensively studied on tumor treatment. Ultrasound can induce both thermal effects and mechanical effects to destroy tumor microenvironment. Purpose：In this study, we investigated cancer tumor treatment by the combination of metformin and therapeutic ultrasound. Materials and Methods：In in-vitro study, CT-26 cancer cells and 96 well dish were used and each well was cultivated with 5 x 103 cells. There were four groups: Control, Met (1mM metformin for 24 hr), pUH (42℃, 0.94 MHz, 0.1 mW and 10% duty cycle ultrasound for 15 min), and pUH+Met. In in-vivo study, BALB/c male mice were used and the mice were injected subcutaneously with CT-26 Luc colon carcinoma cells in the right flank. Treatments were started when the tumors grew up to 50 mm3. The mice were randomly divided into four groups：Control, Met, pUH and Met+pUH. For the metformin treatment, the mice were given a dose of 25mg/kg metformin by oral gavage. For the ultrasound treatment, a frequency of 1 MHz, a duty cycle of 50%, a power of 2.7 W and a duration of 15 min were used. Tumors were measured every two days and H&E staining was used to study the effects of metformin and ultrasound on tumor tissues. Results：In in-vitro study, metformin plus therapeutic ultrasound decreased the cell viability of CT-26 to 60% for a single treatment. It was significantly different as compared to the other three groups. In in-vivo study, the treatment with metformin plus therapeutic ultrasound could effectively suppressed the tumor growth and the experimental results showed that there was significantly different as compared to the other three groups in tumor growth response after four treatments. Lymphocytic infiltration and inflammation could be observed on the H&E staining for the pUH and Met+pUH groups, and tissue rupture was obvious for these two groups. Summary：Metformin combined with therapeutic ultrasound could inhibit cancer cells and tumor growth significantly in the in-vitro and in-vivo studies. H&E staining showed the lymphocytic infiltration, inflammation and tissue disruption, indicating the thermal and mechanical effects of ultrasound sonication on tumor tissues. The results indicate that the effective treatment may come from both the damage by ultrasound sonication and the molecular mechanism caused by metformin. Keywords：Metformin、Therapeutic ultrasound、Tumor growth inhibition1428753 bytesapplication/pdf論文公開時間: 2019/8/24論文使用權限: 同意有償授權(權利金給回饋學校)Metformin超音波熱效應超音波機械效應癌腫瘤抑制Therapeutic ultrasoundTumor growth inhibition[SDGs]SDG3thesis10.6342/NTU201601536http://ntur.lib.ntu.edu.tw/bitstream/246246/277533/1/ntu-105-R03548028-1.pdf