本文關鍵詞： 肝癌 人肝癌細胞SMMC- 橙皮素 Bcl- Bax　出處：《安徽醫(yī)科大學學報》2017年07期 　論文類型：期刊論文

【摘要】：目的以橙皮素為原型,合成了新型化合物HY-12,研究其體外對SMMC-7721細胞的促凋亡作用及機制。方法在3個時間點(24、48、72 h)采用MTT比色法觀察不同濃度HY-12對肝癌細胞、肝細胞增殖的影響,計算半數(shù)抑制率(IC50)。根據(jù)結果選取了24 h,12.5×10-6,25×10-6,50×10-6mol/L 3個濃度進行下述實驗。通過Annexin V/PI雙染,流式細胞儀檢測細胞凋亡率;制備細胞爬片,Hoechst 33342染色,在熒光倒置顯微鏡下觀察細胞凋亡的形態(tài)學改變;采用Western blot法檢測細胞凋亡相關因子Bcl-2、Bax蛋白水平的表達情況。結果 (1)MTT法顯示在對肝細胞活性無顯著影響的前提下,HY-12在體外可抑制SMMC-7721細胞的增殖,且具有劑量-時間依賴性;(2)流式細胞儀檢測顯示HY-12作用后,SMMC-7721細胞凋亡率增加,且呈濃度依賴性趨勢;(3)經(jīng)Hoechst 33342染色,藥物處理后細胞核皺縮斷裂呈現(xiàn)亮藍色新月狀;(4)Western blot結果顯示藥物處理可以使Bax蛋白表達上調(diào),相反地Bcl-2蛋白表達下調(diào)。結論橙皮素衍生物HY-12有明顯抑制肝癌細胞SMMC-7721增殖作用。其機制可能是通過調(diào)節(jié)Bcl-2、Bax表達以誘導細胞凋亡。HY-12可能成為治療肝癌的有效化療藥物。
[Abstract]:Objective to synthesize a novel compound HY-12 from hesperidin, and to study its effect on apoptosis of SMMC-7721 cells in vitro and its mechanism. Methods MTT colorimetric assay was used to observe the effects of different concentrations of HY-12 on the proliferation of hepatoma cells and hepatocytes. The 50% inhibition rate (IC50) was calculated. According to the results, the following three concentrations were selected: 24 h, 12.5 脳 10 -6, 25 脳 10 -6, 50 脳 10 -6 mol / L, respectively. The apoptosis rate was detected by Annexin V / Pi double staining, and the cell climbing slices were prepared for Hoechst 33342 staining. The morphological changes of apoptosis were observed under fluorescence inverted microscope. The expression of apoptosis-related factor Bcl-2 Bax protein was detected by Western blot assay. Results the results showed that HY-12 could inhibit the proliferation of SMMC-7721 cells without significantly affecting the hepatocyte activity in vitro. Flow cytometry analysis showed that the apoptosis rate of SMMC-7721 cells increased after HY-12 treatment, and showed a concentration-dependent trend. Hoechst 33342 staining showed that the apoptosis rate of SMMC-7721 cells was higher than that of SMMC-7721 cells in a dose-dependent manner. The results of Western blot showed that the nuclear shrinkage and cleavage after drug treatment could up-regulate the expression of Bax protein. On the contrary, the expression of Bcl-2 protein was down-regulated. Conclusion HY-12, a derivative of hesperidin, can significantly inhibit the proliferation of SMMC-7721 in hepatocellular carcinoma cells, and its mechanism may be that it may be an effective chemotherapeutic drug for the treatment of hepatocellular carcinoma by regulating the expression of Bcl-2P Bax to induce apoptosis.
【作者單位】： 安徽醫(yī)科大學藥學院安徽醫(yī)科大學肝病研究所安徽省創(chuàng)新藥物產(chǎn)業(yè)共性研究院;
【基金】：安徽省高等學校省級自然科學研究重點項目(編號:KJ2016A364、KJ2016A365) 安徽省自然科學基金面上項目(編號:1408085MKL31) 國家自然科學基金(編號:81273526、81473268)
【分類號】：R96

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