2022 年第 2 期 第 17 卷

高原藏药翼首草提取物对人胃癌BGC-823细胞增殖和凋亡及侵袭的影响

Effect of Tibetan medicine Pterocephalus hookeri extract on proliferation, apoptosis and invasion of human gastric cancer BGC-823 cells

作者：周文星1王晴晴1张成武1刘川川2蒲永强1张晓静1付佳伟1崔桂芳1缪巍1

英文作者：Zhou Wenxing1 Wang Qingqing1 Zhang Chengwu1 Liu Chuanchuan2 Pu Yongqiang1 Zhang Xiaojing1 Fu Jiawei1 Cui Guifang1 Miao Wei1

单位：1青海大学附属医院胃肠肿瘤外科，西宁810001；2青海大学附属医院中心实验室，西宁810001

英文单位：1Department of Gastrointestinal Tumor Surgery the Affiliated Hospital of Qinghai University Xining 810001 China; 2Central Laboratory the Affiliated Hospital of Qinghai University Xining 810001 China

关键词：胃癌BGC-823细胞；翼首草；细胞增殖；细胞凋亡；细胞周期；细胞侵袭

英文关键词：GastriccancerBGC-823cells；Pterocephalushookeri；Cellproliferation；Cellapoptosis；Cellcycle；Cellinvasion

• 摘要：

• 目的  探讨藏药翼首草提取物对人胃癌BGC-823细胞增殖和凋亡及侵袭的影响。方法使用70%甲醇提取翼首草中成分。体外培养人胃癌BGC-823细胞，将不同浓度翼首草提取物作用于BGC-823细胞。使用细胞计数试剂盒-8检测翼首草提取物对BGC-823的半数抑制浓度；流式细胞术检测翼首草提取物作用后细胞凋亡和细胞周期情况；荧光定量聚合酶链反应分析翼首草提取物作用后细胞增殖细胞核抗原(PCNA)、细胞周期蛋白D1(Cyclin D1)mRNA表达水平；蛋白质印迹法分析翼首草提取物作用后细胞PCNA、Cyclin D1和活化的含半胱氨酸的天冬氨酸蛋白水解酶3（cleaved-Caspase-3）蛋白表达水平；Transwell实验分析翼首草提取物作用后细胞侵袭能力。结果  翼首草提取物处理的胃癌BGC-823细胞活力下降，且随着药物浓度的增加抑制作用越明显，作用24 h和48 h对胃癌BGC-823细胞抑制作用的半数抑制浓度分别为（30±8）μg/ml和（21±4）μg/ml，本研究后续分别以10、20、40 μg/ml翼首草提取物处理胃癌BGC-823细胞48 h进行实验。10、20、40 μg/ml翼首草提取物组胃癌BGC-823细胞凋亡率均明显高于0 μg/ml组［（13.13±2.12）%、（13.15±1.29）%、（20.24±0.62）%比（2.91±0.96）%］，细胞G0/G1期比例明显高于0 μg/ml组［（65.0±2.3）%、（68.2±2.9）%、（73.1±1.4）%比（58.1±2.8）%］，PCNA和Cyclin D1 mRNA表达水平均低于0 μg/ml组［PCNA mRNA：（0.56±0.16）、（0.34±0.15）、（0.22±0.12）比（1.00±0.30）；Cyclin D1 mRNA：（0.38±0.15）、（0.33±0.10）、（0.28±0.12）比（1.00±0.26）］（均P＜0.05）。20、40 μg/ml翼首草提取物组BGC-823细胞PCNA和Cyclin D1蛋白表达水平和迁移细胞数均低于0 μg/ml组，cleaved-Caspase-3蛋白表达水平均高于0 μg/ml组（均P＜0.05）。结论  翼首草提取物处理能抑制人胃癌BGC-823细胞增殖和侵袭，并促进细胞凋亡的发生。

• Objective To explore the effect of Tibetan medicine Pterocephalus hookeri extract on the proliferation, apoptosis and invasion of human gastric cancer BGC-823 cells. Methods Ingredients in Pterocephalus hookeri were extracted by 70% methanol. Human BGC-823 cells were cultured in vitro, and different concentrations of Pterocephalus hookeri extraction were applied to BGC-823 cells. Cell counting kit-8 was used to detect the half-inhibitory concentration (IC50) of Pterocephalus hookeri extract on BGC-823. Flow cytometry was used to detect the cell cycle and apoptosis after Pterocephalus hookeri treatment. The expression levels of proliferating cell nuclear antigen (PCNA) and Cyclin D1 mRNA were analyzed by fluorescence quantitative polymerase chain reaction. Western blotting was used to analyze protein expression level of PCNA, Cyclin D1 and cleave-Caspase-3 in cells after Pterocephalus hookeri extract treatment. The cell invasion ability after Pterocephalus hookeri extract treatment was analyzed by Transwell test. Results The viability of BGC-823 cells treated with Pterocephalus hookeri extract decreased, and the inhibitory effect became more obvious with the increase of drug concentration. The IC50 of inhibitory effect on BGC-823 cells for 24 h and 48 h was （30±8）μg/ml and （21±4）μg/ml, respectively. The experiment was conducted on the treatment of gastric cancer BGC-823 cells 48 h by Pterocephalus hookeri extract 10, 20 and 40 μg/ml. The apoptotic rate of BGC-823 cells in groups of Pterocephalus hookeriextract 10, 20 and 40 μg/ml were significantly higher than that in group of 0 μg/ml［（13.13±2.12）%,（13.15±1.29）%,（20.24±0.62）% vs （2.91±0.96）%］, the proportion of cell G0/G1 phase were significantly higher than that in group of 0 μg/ml［（65.0±2.3）%, （68.2±2.9）%, （73.1±1.4）% vs （58.1±2.8）%］, the expression levels of PCNA and cyclin D1 mRNA were lower than those in group of 0 μg/ml［PCNA mRNA：（0.56±0.16）, （0.34±0.15）, （0.22±0.12） vs （1.00±0.30）；Cyclin D1 mRNA：（0.38±0.15）, （0.33±0.10）, （0.28±0.12） vs （1.00±0.26）］ (all P＜0.05). The expression levels of PCNA, cyclin D1 protein and the number of migrating cells in BGC-823 cells in groups of 20 and 40 μg/ml were lower than those in group of 0 μg/ml, and the expression level of cleaved-Caspase-3 protein was higher than that in group of 0 μg/ml (all P＜0.05). Conclusion Pterocephalus hookeriextract  inhibits the proliferation and invasion of human gastric cancer BGC-823 cells and promotes the occurrence of cell apoptosis.