近年來，癌症是儼然成為最頭痛的問題，它可以促進血管新生以
及讓癌細胞轉移，讓癌症如僵屍一般，還是沒有任何有效辦法可以徹
底根除。
本實驗室從 Lactobacillus rhamnosus DSM 6594 上搜索其定額感
應(Quorum sensing)裡的 HPK 蛋白，以此為模板再以融合引物與巢式
聚合酶連鎖反應(FPNI-PCR)做外部擴增探索未知序列，比對資料庫
後，篩選出 MKK40 進行後續對抗人類大腸癌細胞 SW480 的測試。
MTT 結果發現，MKK40 會誘發 SW480 死亡，IC50為 34.2 μg/ml。流
式細胞儀結果顯示，MKK40 會觸發 SW480 走向細胞凋零以及壞死的
路徑，再利用西方點墨法測定 MKK40 對 SW480 的死亡路徑相關蛋
白，並用共軛焦顯微鏡觀察 MKK40 進入細胞後的外觀和位置上的變
化。
綜合各實驗結果，MKK40 可以誘導 SW480 啟動細胞凋零以及壞
死的機制，並且不會隨著時間的增加進入到細胞內，而是繼續停留在
細胞膜上。

Recently, Cancer is still the biggest problem what the human being
haven’t solved. Cancer cell has many functions like: metastasis and
angiogenesis. It can gain nutrient from the new-born vessel and diffuse to
other organism. In briefly, whether we develop a new drug for cancer, it
also develop.
Our lab dedicated to research new drug for human colon caner cell
SW480 such as BD21, KL15…etc. These drugs we find out have great
activity to SW480. Now we search from the bacteriocin database and
find out the new drug, MKK40. In CCK-8 cell viability test, we find that it has good activity to anti-cancer cell SW480. IC50 of SW480 is 34.2μg/ml.
Further, we use flow cytometry figure out when we treat MKK40 in
SW480 cell. It can trigger SW480 to proceed apoptosis and necrosis.In
western blot experiment, we use Pro-Caspase 3, Caspase 7, Caspase 9 and
TNFR1 to determine whether these apoptosis marker is relate to MKK40
induce. And the necrosis biomarker: RIP3, MLKL and Cyclophilin A is
used to determine. We also track MKK40 position after incubate by
different time( 30min, 1hr, 6hr, 12hr, 24hr) in SW480.
In summary, MKK40 trigger both apoptosis and nercrosis in SW480
cell and no matter how long MKK40 treated, it still attached in cell
membrane.

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