桦褐孔菌水提物通过caspase 3介导细胞凋亡对口腔鳞癌抑制作用的研究

目的 探讨桦褐孔菌水提物对口腔鳞癌SCC - 25细胞的抑制作用及其相关机制。方法 使用水提醇沉方法获得桦褐孔菌水提物，并以不同浓度加至SCC - 25细胞，根据对细胞增殖和凋亡的作用选择合适剂量用于后续实验。将SCC - 25细胞分为空白组、caspase 3抑制剂组、si - NC组和si - caspase 3组，分别加入caspase 3抑制剂或通过脂质体转染法将siRNA转染入细胞。采用CCK8法和集落形成实验检测桦褐孔菌水提物对SCC - 25细胞增殖能力的影响，通过流式细胞术和彗星实验检测细胞凋亡和DNA损伤情况，使用Western blot检测细胞caspase 3、caspase 7、B细胞淋巴瘤/白血病 - 2（B - cell lymphoma / leukemia 2，bcl - 2）蛋白表达情况。实验结果按t检验和one - way ANOVA统计分析。结果 桦褐孔菌水提物呈剂量依赖性提高caspase 3蛋白表达，诱导细胞DNA损伤，促进细胞凋亡并抑制SCC - 25细胞增殖（P＜0.05）。在基因水平沉默caspase 3或使用caspase 3抑制剂能够拮抗桦褐孔菌水对SCC - 25细胞的促凋亡和抑制增殖作用（P＜0.05）。结论 桦褐孔菌水提物通过caspase 3介导SCC - 25细胞DNA损伤和细胞凋亡，抑制口腔鳞癌细胞的增殖。

Study on the inhibitory effect of water extracts of Inonotus obliquus on oral squamous cell carcinoma through caspase 3-dependent apoptosis

Objective To investigate the inhibitory effect of water extracts of Inonotus obliquus on oral squamous cell carcinoma SCC-25 cells and its related mechanism. Methods The aqueous extract of Inonotus obliquus was obtained using the aqueous alcoholic precipitation method and added to SCC-25 cells at different concentrations, and the appropriate doses were selected for subsequent experiments based on the effects on cell proliferation and apoptosis. SCC-25 cells were divided into blank group, caspase 3 inhibitor group, si-NC group and si-caspase 3 group, caspase 3 inhibitor was added and the siRNAs were transfected into each group by liposome transfection method, respectively. The effect of aqueous extract of Inonotus obliquus on the proliferation ability of SCC-25 cells was examined by CCK8 method and colony formation assay, apoptosis and DNA damage were detected by flow cytometry and comet assay, and the protein expression of caspase 3, caspase 7 and bcl-2 were detected by western blot. The experimental results were statistically analyzed by t test and one-way ANOVA. Results The aqueous extract of Inonotus obliquus increased the protein expression of caspase 3, induced cellular DNA damage, promoted apoptosis and inhibited the proliferation of SCC-25 cells in a dose-dependent manner (P<0.05). Silencing caspase 3 at the gene level or using caspase 3 inhibitors antagonized the pro-apoptotic and inhibitory proliferative effects of aqueous extract of Inonotus obliquus on SCC-25 cells (P<0.05). Conclusion Aqueous extract of Inonotus obliquus inhibits the proliferation of oral squamous carcinoma cells through caspase 3-mediated DNA damage and apoptosis in SCC-25 cells.