目的 研究大黃素對(duì)人肝癌Huh7細(xì)胞的凋亡作用及分子機(jī)制。方法 培養(yǎng)肝癌Huh7細(xì)胞,1、3、10、30、100 μmol/L的大黃素處理24 h,5-氟尿嘧啶(5-FU)作為陽(yáng)性藥,MTT法檢測(cè)細(xì)胞存活率;30 μmol/L大黃素處理細(xì)胞0、3、6、24 h,光學(xué)顯微鏡觀察細(xì)胞形態(tài)變化;Annexin V-FITC/PI雙染法與流式細(xì)胞術(shù)檢測(cè)細(xì)胞凋亡,Western blotting檢測(cè)細(xì)胞凋亡相關(guān)蛋白的表達(dá)變化。結(jié)果 大黃素顯著抑制Huh7細(xì)胞生長(zhǎng)且呈濃度依賴性,其IC₅₀值為11.55 μmol/L。30 μmol/L大黃素處理細(xì)胞,隨著藥物作用時(shí)間的增加,細(xì)胞明顯固縮和凝聚,大量細(xì)胞脫離培養(yǎng)皿底部;細(xì)胞凋亡明顯;p-Akt、Bcl-2蛋白表達(dá)量減少,cleaved caspase-3蛋白表達(dá)量增加。結(jié)論 大黃素通過(guò)Akt信號(hào)途徑誘導(dǎo)肝癌Huh7細(xì)胞凋亡。

Objective To elucidate the effect of emodin on inducing apoptosis of humam hepatoma Huh7 cells and its molecular mechanism. Methods Huh7 cells were treated with 1, 3, 10, 30 and 100 mol/L emodin for 24 h, 5-FU as positive drug. The viability of Huh7 cells was detected by MTT assay. Cells were treated with 30 μmol/L emodin for 0, 3, 6 and 24 h, and the morphological changes of cells were observed by optical microscope. The apoptosis of Huh7 cells were determined by Annexin V-FITC/PI double staining and flow cytometer. The expression levels of Akt signalling pathway proteins were determined by Western blotting. Result Emodin could significantly inhibit the proliferation of Huh7 cells in a dose-dependent manner. The IC₅₀ values were determined as 11.55 μmol/L. Cells were treated with 30 mol/L emodin, with the increasing of the time, cells significantly reduced and condensed, a large number of cells droped from the bottom of the culture dish; Emodin could induce apoptosis; emodin could significantly down-regulated p-Akt, Bcl-2 protein expression levels while up-regulated the cleaved caspase-3 expression levels in Huh7 cells. Conclusion These findings suggested that emodin can induce mitochondrial-related apoptosis on Huh7 cells by inhibiting Akt activity.

黑龍江省博士后科研啟動(dòng)項(xiàng)目(LBH-Q13132);黑龍江省自然科學(xué)基金資助項(xiàng)目(LC2015036);學(xué)成、引進(jìn)人才科研啟動(dòng)計(jì)劃(XYB2013-24)