目的 探究白屈菜紅堿（CHE）對腺樣囊性癌細胞（ACC2）生長的抑制作用及機制。方法 利用CCK8法、EdU法、Hoechst33342/PI雙染色法、試劑盒法檢測CHE對ACC2細胞活力、細胞增殖、細胞凋亡和活性氧（ROS）水平的影響；通過Western blotting技術檢測CHE對Cleaved-Caspase 3、PARP、NF-κB、p-JNK、p-p38蛋白表達的影響；利用斑馬魚移植瘤模型檢測CHE對斑馬魚體內(nèi)ACC2細胞生長的抑制作用。結果 CCK-8結果顯示：與對照組比較，2、3、4、5、6、7、8、9、10 μmol/L的CHE顯著降低ACC2細胞的存活率（P<0.05、0.01），且呈濃度相關性； ROS檢測結果顯示：與對照組比較，5、8 μmol/L的CHE導致ACC2細胞內(nèi)的ROS水平顯著上升（P<0.05、0.01）； EdU增殖檢測結果表明：與對照組比較，5、8 μmol/L的CHE致使ACC2細胞的增殖能力顯著下降（P<0.01）；Hoechst/PI染色結果顯示：與對照組比較，CHE 5、8 μmol/L組ACC2細胞凋亡率顯著上升（P<0.01）。抗氧化劑N-乙酰半胱氨酸（NAC）顯著抑制CHE誘導的ROS水平升高、細胞凋亡增加（P<0.01）；Western blotting結果顯示：2、5、8 μmol/L的CHE能夠顯著上調(diào)Cleaved-Caspase 3、PARP、NF-κB蛋白的表達（P<0.01），且呈現(xiàn)濃度相關性，5、8 μmol/L的CHE能夠顯著上調(diào)p-JNK的蛋白表達（P<0.01），8 μmol/L的CHE能夠顯著上調(diào)p-p38的蛋白表達（P<0.01）；NAC顯著降低由CHE導致的Cleaved-Caspase 3、PARP、NF-κB、p-JNK、p-p38蛋白表達增加（P<0.01），5、8 μmol/L CHE能夠有效抑制斑馬魚體內(nèi)腫瘤的生長（P<0.01）。結論 體外及斑馬魚移植瘤模型證明，CHE可以有效抑制ACC2細胞生長，其機制與提高細胞ROS水平，上調(diào)NF-κB、p-JNK、p-p38表達，從而抑制細胞增殖、誘導細胞凋亡相關。

Objective To investigate the inhibitory effect of chelerythrine (CHE) on adenoid cystic carcinoma (ACC2) cells and its molecular mechanism. Methods CCK8, EdU, hoechst33342/PI double staining, and test kit method were used to detect the effects of CHE on ACC2 cell viability, cell proliferation, apoptosis and reactive oxygen species (ROS) level. The effects of CHE on the expression levels of Caspase-3, PARP, NF-κB, P-JNK and p-p38 were detected by Western blotting. The inhibitory effect of chelerythrine on the growth of ACC2 cells was detected by zebrafish xenograft tumor model. Results CCK-8 assay showed that: CHE can significantly inhibited the proliferation of ACC2 cells in a concentration dependent manner (P < 0.05 and 0.01) at the concentration of 2, 3, 4, 5, 6, 7, 8, 9, 10 μmol/L. The results showed that ROS in ACC2 cells was significantly increased at the concentration of 5 and 8 μmol/L (P < 0.05 and 0.01). EdU assay results showed that the proliferation of ACC2 cells was significantly decreased by CHE at the concentration of 5 and 8 μmol/L (P < 0.01). Hoechst/PI staining showed that apoptosis was significantly induced by CHE at the concentration of 5 and 8 μmol/L (P < 0.01). while N-acetylcysteine (NAC) significantly inhibited the increase of ROS and apoptosis induced by CHE (P < 0.01). Western blotting results showed that the expression of Cleaved Caspase 3, PARP and NF-κB were significantly up-regulated by 2, 5, and 8 mol/L CHE (P < 0.01), the expression of p-JNK was significantly up-regulated by 5 and 8 mol/L CHE (P < 0.01), and the protein expression of p-p38 was significantly up-regulated by 8 mol/L CHE (P < 0.01). NAC significantly decreased the expression of Cleaved Caspase 3, PARP, NF-κB, p-JNK, p-p38 protein induced by CHE (P < 0.01). CHE of 5 and 8 mol/L could effectively inhibit the growth of ACC2 cells in zebrafish (P < 0.01). Conclusion CHE can effectively inhibit the proliferation of ACC2 cells in vivo and zebrafish xenograft tumor model. Elevation of the ROS level induced by CHE, upregulation of NF-κB, p-JNK and p-p38 and then lead to the inhibition of proliferation and induction of apoptosis were involved in the inhibition of ACC2 cells in vivo and in vivo.

國家重點研發(fā)計劃（2018YFC1707300）；山東省重大科技創(chuàng)新工程項目（2019JZZY020905）；齊魯工業(yè)大學（山東省科學院）科教產(chǎn)融合創(chuàng)新試點工程項目（2020KJC-ZD08）