胰臟癌是現有人類罹患的惡性腫瘤中，最具侵略性、致死性和預後不佳的一種癌症。胰臟癌的病患平均五年存活率約只有百分之六，雖然標準治療藥物gemcitabine對部分的胰臟癌病患有療效，但在晚期胰臟癌病患中平均只能維持5.6個月的存活時間，因此發展更多新型治療策略來控制此棘手的癌症是十分重要的。隨著癌症研究的精進，已有許多報導顯示過度的Aurora kinase表現在胰臟癌的發展過程中扮演關鍵的角色；而異常的mTOR kinase訊息活化也與胰臟癌的癌化過程和抗藥性息息相關；另外，也有研究指出過度活化TrkA kinase訊息傳導路徑與胰臟癌的高度轉移性有一定關聯。因此本研究的目的，是去探討一新穎藥物BPR1K-X對於胰臟癌細胞是否有抑癌作用，並了解其分子機制。首先在試管內激脢活性分析中，BPR1K-X可有效抑制Aurora A kinase、Aurora B kinase與TrkA kinase的活性。而在細胞實驗中，BPR1K-X對四株不同的胰臟癌細胞皆有療效。本研究進一步利用西方墨點法，也證實BPR1K-X可藉由抑制Aurora A kinase、Aurora B kinase、TrkA與mTOR的訊息傳導路徑，進而抑制癌細胞的生長。流式細胞儀的分析則指出BPR1K-X會造成細胞周期停留G2/M期，使癌細胞無法正常分裂進而產生多倍體。另外在西方墨點法、免疫螢光染色以及細胞自噬作用抑制的實驗中，都顯示BPR1K-X會增加細胞凋亡與細胞自噬作用造成癌細胞的死亡。總結以上結果，BPR1K-X為有潛力的多分子抑制劑，未來有機會應用為治療胰臟癌的新抗癌藥物。

Pancreatic cancer is widely known as the most aggressive solid malignancies with poor prognosis. The overall 5-year survival is 6%, and the standard treatment with gemcitabine only yields a median survival of 5.6 months in advanced disease. Therefore, developing novel therapeutic approaches is desperately needed. Previous studies have shown that overexpression of oncogenic Aurora kinases play a critical role in pancreatic carcinogenesis. Aberrant mTOR signaling in cancer cells is also associated with the drug resistance and tumorigenic potential. Besides, recent studies reveal that TrkA plays an important role in pancreatic cancer development, making these signaling pathways as important anticancer targets. In this study, we aim to elucidate the anticancer activity of a novel multiple kinase inhibitor, BPR1K-X, in human pancreatic cancer cell lines. In vitro kinase inhibition assay indicated that BPR1K-X inhibited multiple kinase activity such as Aurora A and B, and TrkA at low nano-molar concentrations. Results of the MTT cell viability assay, BrdU cell proliferation assay and LDH cytotoxicity assay showed that BPR1K-X was effective in targeting the human AsPC-1, MIA PaCa-2, BxPC-3, and PANC-1 pancreatic cancer cells in vitro. At the molecular level, Western blot analysis revealed that BPR1K-X suppressed p-Aurora A and B, p-mTOR, and p-TrkA expression in pancreatic cancer cells. In addition, Western blot analysis and flow cytometric analysis revealed that BPR1K-X decreased the expression of cyclin B1 and induced endo-reduplication by the formation of polyploidy, respectively. It also modulated autophagy and induced apoptosis in the treated pancreatic cancer cells. Taken together, our study suggests that BPR1K-X is a potent multiple kinase inhibitor that is able to target human pancreatic cancer cells.

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