【摘要】：目的：觀察姜黃素（Curcumin）誘導人髓母細胞瘤Daoy細胞自噬的作用，并探討其可能機制。 方法：體外培養(yǎng)人髓母細胞瘤Daoy細胞，透射電鏡檢測姜黃素處理前后Daoy細胞超微結構變化，免疫熒光染色、RT-PCR和Westernblot檢測自噬相關蛋白Beclin1和LC3的表達；Western blot檢測PI3K/Akt/mTOR信號通路中PI3K、p-Akt和p-mTOR的表達；姜黃素分別與自噬抑制劑wortmannin、PI3K激動劑insulin聯(lián)合處理細胞后，Western blot檢測自噬相關蛋白Beclin1和LC3及PI3K/Akt/mTOR信號通路相關蛋白的表達變化。 結果：透射電鏡結果顯示，隨姜黃素作用時間增加，自噬體明顯增多。免疫熒光染色顯示Beclin1和LC3在姜黃素處理組中的表達強度明顯高于對照組。RT-PCR顯示與對照組相比，Beclin1mRNA和LC3mRNA的表達在處理組(24h和48h)明顯增強，差異有統(tǒng)計學意義（F=1360.962，42.366，P 0.05）。Western blot顯示處理組（24h和48h）Beclin1、LC3-Ⅱ/LC3-Ⅰ的表達明顯高于對照組（0h）,而PI3K、p-Akt和p-mTOR的表達明顯低于對照組，差異有統(tǒng)計學意義（F=32.723,11.847，，329.662,80.638，183.536，P 0.05）。自噬抑制劑wortmannin與姜黃素聯(lián)合處理后不能降低姜黃素誘導的自噬。PI3K激動劑insulin與姜黃素聯(lián)合處理后不能有效逆轉姜黃素對PI3K/Akt/mTOR信號通路的抑制作用。 結論：姜黃素通過誘導自噬抑制髓母細胞瘤Daoy細胞生長,其機制可能與上調自噬相關基因表達，抑制自噬負調控PI3k/Akt/mTOR信號通路有關。
[Abstract]:Aim: to investigate the effect of curcumin (Curcumin) on autophagy of human myeloblastoma Daoy cells and its possible mechanism. Methods: human myeloblastoma Daoy cells were cultured in vitro. Ultrastructural changes of Daoy cells before and after curcumin treatment were detected by transmission electron microscope. The expression of autophagy associated protein Beclin1 and LC3 was detected by RT-PCR and Westernblot. Western blot was used to detect the expression of PI3KP- Akt and p-mTOR in PI3K/Akt/mTOR signaling pathway, and curcumin combined with wortmanninnine PI3K agonist insulin was used to detect the expression of Beclin1, LC3 and PI3K/Akt/mTOR signal pathway related proteins. Results: the results of transmission electron microscope showed that the autophagy increased with the increase of curcumin action time. Immunofluorescence staining showed that the expression of Beclin1 and LC3 in curcumin treated group was significantly higher than that in control group. RT-PCR showed that the expression of Beclin1 mRNA and LC3mRNA in the treatment group (24h and 48h) was significantly higher than that in the control group. The expression of Beclin1 + LC3- 鈪

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