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目的观察天然植物冬凌草提取物冬凌草甲素对结肠癌LoVo细胞增殖、凋亡的作用,并分析其可能的作用机制。方法体外培养LoVo细胞,用不同浓度冬凌草甲素(0-240μmol/L)分别处理24,48,72h,MTT法检测细胞活力的改变。冬凌草甲素(终浓度为0,30,60,120μmol/L)分别处理LoVo细胞48h,流式细胞仪检测细胞凋亡率的变化,ELISA法检测细胞上清液中PGE2的含量;Westernblot检测细胞蛋白环氧合酶-2(COX-2)、p-NF-κB(p65)、Bcl-2及Bax的表达的改变。结果冬凌草甲素(15-240μmol/L)能显著抑制LoVo细胞活力(P<0.05),呈剂量、时间依赖性。冬凌草甲素(终浓度为0,30,60,120μmol/L)处理LoVo细胞48h后,细胞凋亡率分别为8.83%,15.5%,22.64%和25.86%;冬凌草甲素处理LoVo细胞48h后,细胞Bax蛋白表达上调,Bcl-2蛋白表达下调(P<0.05),同时p-NF-κB(p65)、COX-2蛋白的表达及PGE2合成降低(P<0.05)。结论冬凌草甲素可通过抑制p-NF-κB、COX-2表达及PGE2合成,继而上调促凋亡蛋白Bax表达,下调抗凋亡蛋白Bcl-2表达,抑制LoVo细胞增殖并诱导细胞凋亡。
Objective To observe the effect of Rubescens extract, a natural plant extract, on the proliferation and apoptosis of colon cancer LoVo cells and to analyze its possible mechanism. Methods LoVo cells were cultured in vitro. The cells were treated with different concentrations of oridonin (0-240μmol / L) for 24, 48 and 72 hours respectively. The cell viability was measured by MTT assay. Lovastatin A (0, 30, 60, 120μmol / L) was used to treat LoVo cells for 48h respectively. The apoptosis rate of LoVo cells was detected by flow cytometry. The content of PGE2 in supernatant was detected by ELISA. Changes in the expression of cellular protein cyclooxygenase-2 (COX-2), p-NF-κB (p65), Bcl-2 and Bax. Results Oridonin (15-240μmol / L) significantly inhibited the viability of LoVo cells (P <0.05) in a dose-and time-dependent manner. After treatment with oridonin (0, 30, 60 and 120μmol / L) for 48h, the apoptosis rates of LoVo cells were 8.83%, 15.5%, 22.64% and 25.86% After 48h, the expression of Bax protein was up-regulated and the expression of Bcl-2 protein was down-regulated (P <0.05). Meanwhile, the expression of p65, COX-2 protein and PGE2 decreased. Conclusion Oridonin can up-regulate the expression of Bax, decrease the expression of anti-apoptotic protein Bcl-2, inhibit the proliferation of LoVo cells and induce the apoptosis by inhibiting the expression of p-NF-κB, COX-2 and PGE2 Death.