肝細胞肝癌是一種死亡率非常高的癌症,它的致死率是再癌症中排名第三位. 常見的治療方式有手術切除和經動脈栓塞,而且一般常見的化療藥物有epirubicin, doxorubicin, cisplatin and 5-FU, 但是治療效果往往不是很好,而且化療藥物對患者會產生極大的副作用, 例如: 掉頭髮,嘔吐. 這些化療藥物在臨床上沒有很好的抗肝癌效果, 在我們之前的研究中我們發現epirubicin對大鼠的原位肝癌的腫瘤大小有抑制的效果,但是我們發現epirubicin對大鼠本身會造成極大的副作用,例如: 掉毛, 體重下降, 脾臟變小, 白血球數目下降, GOT上升. 所以我們想要利用一些低毒性的臨床用藥來治療原位肝癌,像celecoxib就是一種毒性低的臨床用藥,它本身是一個第二型環氧化酵素 (COX-2) 的抑制劑,而且毒性也比傳統的非類固醇抗發炎藥物來的低, 而且專一性非常的高, 而且在我們的實驗中我們發現N1-S1肝癌細胞株和Clone 9正常肝細胞株中所測得的半抑制劑量是100 microM, 而且我們發現N1-S1肝癌細胞株的 COX-2蛋白質表現量和mRNA表現量是非常低的, 所以N1-S1細胞是一種COX-2表現量非常低的肝癌細胞株, 但有趣的是我們發現celecoxib在細胞實驗中可以經由COX-2 independent的作用機轉去正調控PPAR-gamma和PTEN的表現，進而使得AKT的磷酸化比例下降，而且我們每天餵食celecoxib (30mg/kg/rat)且連續餵食七天,治療過後我們不管用超音波還是電腦斷層掃描, 我們都可以看出celecoxib在活體中對肝腫瘤我抑制的效果, 而且我們發現治療組大鼠的肝組織中CD-31有變少的趨勢,這也代表celecoxib在活體中具有抑制腫瘤血管新生的效果，在者，我們發現celecoxib在活體中可以減少Ki-67的表現量，這代表癌細胞的增生速度減緩，以及看見治療組中的TUNEL增加，這表示癌細胞細胞凋亡的比例提高。在免疫系統方面，我們發現治療組可以減少FOXP3在腫瘤週遭的比例，所以celecoxib有破壞癌細胞逃脫免疫系統機轉的潛力 除此之外, 在動物實驗中我們發現celecoxib對動物本身沒有明顯的副作用,所以我們認為celecoxib是一個適合與epirubicin或是其他化療藥物組合使用的輔助抗肝腫瘤藥劑。

Hepatocellular carcinoma (HCC) is one of deadliest cancers worldwide and ranking the third among all cancer-related mortalities. Current effective therapeutic approaches for HCC include surgical resection and trans-arterial embolization (TAE). Chemotherapy remains largely ineffective, and most popular used agents are epirubicin, doxorubicin, cisplatin and 5-FU. Besides, these chemotherapic drugs had potential serious side-effects such as low blood count, hair loss, vomiting, and they rarely present good anti-HCC effect in clinical practice. Our previous studies found that epirubicin injection attenuated the tumor burden of orthotopic Novikoff hepatoma, but caused serious side effects to hosts including reduction in spleen weight, white count, and body weight and high GOT level. Therefore, we aimed to evaluate possible alternative treatment such as COX-2 inhibitor for HCC. Celecoxib is a highly selective COX-2 inhibitor and less toxic than the traditional non-selective NSAIDs. Celecoxib showed relatively low cytotoxicity in Novikoff N1-S1 hepatoma cells and Clone 9 normal hepatocytes with an IC50 of up to 100 microM. Expression analysis revealed that COX-2 expression is very low in N1-S1 cells at protein and mRNA levels. Thus, N1-S1 is a kind of hepatoma cell line with low COX-II level. Interestingly, celecoxib upregulated PTEN expression and decreased AKT phosphorylation in vitro by COX-2 independent pathway, and then oral administration of celecoxib (30 mg/kg) for 7 days showed tendency of tumor suppression of Novikoff hepatoma in rats revealed by ultrasound and computed tomography (CT) scan. Histological analysis revealed that CD31-positive neo-vascularization、Ki-67-positive cell-proliferation and FOXP3-positive regulatory T cells were found to reduce in celecoxib-treated rats, and then TUNEL-positive apoptotic cells were found to increase in celecoxib-treated rats. Besides, celecoxib-treated rats exhibited no significant side effect. Therefore, oral celecoxib may be a suitable chose of adjuvant therapy in combination with epirubicin or other chemotherapeutic agents for the treatment of HCC.

Abbreviations 4
Abstract in Chinese 5~6
Abstract in Englis 7~8
Introduction 9~11
Materials and Methods 12~17
Results 18~25
Discussion 26~28
References 29~34
Figures and Legends 35~58
Appendix 59~64

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