ROGDI是一個未知功能的基因，此基因位於人類第16對染色體上，遺傳編碼區全長為864鹼基對，其蛋白質由287個胺基酸組成，在許多動物中具有保守性。根據NCBI資料庫的預測，ROGDI蛋白具有白胺酸拉鍊功能區。在本實驗室先前的研究中，人類ROGDI會影響不同細胞株的增生速度，另外ROGDI也在動物模式肝纖維化過程中增加表現，並參與肝星狀細胞的活化，影響其增生、移行的能力。而在腦癌細胞株T98G與U251中也發現到，過量表現ROGDI的細胞，p53與p27的mRNA表現量上升。本研究中分別使用癌症抑制基因p53野生型與缺失型的肝癌細胞株，探討細胞過量表現ROGDI時p53的表現情形，與對於處理抗癌藥物的影響，並比較兩者和細胞凋亡的關聯。從結果得知ROGDI與p53的基因表現能影響在肝癌細胞株處理抗癌藥物的反應，並促使其進入細胞凋亡。

ROGDI was a novel gene with unknown function, located on human chromosome 16p13.3. The coding region of the gene is 864 bp that encodes 287 amino acids. According to GenBank database, ROGDI contains leucine zipper domain. Previous studies in our laboratory showed that ROGDI increases cell proliferation in cell lines. In addition, overexpression of ROGDI induces p53 and p27 mRNA levels in human glioma cell line T98G and U251. In this study, we use two hepatocellular carcinoma cell lines, Hep G2 and Hep 3B, which contains wild-type and deleted tumor suppressor protein p53 respectively to investigate the expression of p53 and the response of anticancer drugs treatment in ROGDI overexpression cells. In addition, we compare the relation between the cell apoptosis the expression of p53 and ROGDI. Hence, we found that expression of p53 and ROGDI influences the cell response to anticancer drugs and induces apoptosis.

中文摘要…………………………………………………………………I
Abstractt….………………………………………………………………II
目錄…………………………………………………………………..…III
壹、前言…………………………………………………………………1
一、ROGDI……………………………………………………...…1
二、5-氟尿嘧啶……………………………………………………3
三、順鉑……………………………………………………………4
四、Etoposide………………………………………………………5
五、Doxorubicin………………………………………..……….….6
六、p53蛋白…………………………………………………….…7
七、p27蛋白………………………………...………………….….8
八、p21蛋白………………………………………………….……8
九、細胞凋亡………………………………………………….…...9
貳、實驗目的……………………………………………………………11
參、材料與方法…………………………………………………………12
一、小量質體製備………………………………………………...12
二、構築誘導式表現載體pBIG2i……….……………………….15
三、細胞培養……………………………………………………...16
四、細胞轉染………………………………………………………18
五、抗癌藥物的處理………………………………………………18
六、Total RNA的萃取……………………………………….……19
七、反轉錄作用……………………………………………….…..20
八、聚合酶連鎖反應……………………………………………...21
九、瓊膠電泳分析…………………………………………..…….21
十、西方墨點法…………………………………………………...22
十一、細胞生長曲線……………………………………………... 26
十二、以瓊脂凝膠電泳偵測細胞凋亡…………………………...26
十三、流式細胞儀分析…………………………………………...27
肆、結果……………………………………………….………………..29
伍、討論………………………………………………………………...36
陸、參考文獻…………………………………………………………...40
圖表…………...……………………………………………..……….…48
附錄………………………………………………………………….….65

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