中国普外基础与临床杂志

中国普外基础与临床杂志

氧化苦参碱诱导胃癌 BGC-823 细胞发生 caspase-12 依赖性内质网应激凋亡的机制研究

查看全文

目的 研究氧化苦参碱对胃癌 BGC-823 细胞增殖和凋亡的影响,并探讨内质网应激参与氧化苦参碱诱导胃癌细胞的凋亡情况并阐明其机制。 方法 将对数生长期的胃癌 BGC-823 细胞分为对照组、单药组(用 10、30、60 和 90 μmol/L 浓度的氧化苦参碱处理)和联合组(各浓度的氧化苦参碱联合 2 μmol/L 内质网应激抑制剂 salubrinal 处理)。用 MTT 法观察氧化苦参碱对胃癌 BGC-823 细胞生长的抑制作用,用流式细胞仪分析氧化苦参碱诱导胃癌 BGC-823 细胞凋亡及其对细胞周期的影响,用 Western blot 和 RT-PCR 法分别检测内质网应激标志分子 GRP78/Bip 和内质网应激介导凋亡分子 caspase-12 蛋白和基因的表达情况。 结果 ① 与对照组比较,氧化苦参碱能明显抑制胃癌 BGC-823 细胞生长且呈浓度-时间依赖性(P<0.05),其 IC50(48 h)值为(59.5±0.5)μmol/L;与相应浓度单药组比较,30、60 和 90 μmol/L 氧化苦参碱联合组对细胞的抑制作用明显减弱(P<0.05)。② 流式细胞仪分析结果显示,氧化苦参碱能浓度依赖性诱导胃癌 BGC-823 细胞凋亡和 G2/M 期细胞阻滞(P<0.05);与相应浓度单药组比较,60 μmol/L 氧化苦参碱联合组处理胃癌 BGC-823 细胞 48 h 后明显提高细胞凋亡率和增加 G2/M 期细胞数(P<0.05)。③Western blot 和 RT-PCR 法检测结果显示,与对照组比较,随着氧化苦参碱浓度的增加,单药组 GRP78/Bip、caspase-12 蛋白及基因水平呈明显增强的趋势(P<0.05);与相应浓度单药组比较,60 及 90 μmol/L 氧化苦参碱联合组胃癌 BGC-823 细胞中 GRP78/Bip、caspase-12 蛋白及基因表达水平均明显降低(P<0.05)。 结论 氧化苦参碱对人胃癌 BGC-823 细胞有明显的生长抑制作用,该作用可能与 caspase-12 依赖性诱导凋亡及上调 GRP78/Bip 表达水平有关,该结论需要进一步实验验证。

Objective To study effects of oxymatrine on proliferation and apoptosis of gastric cancer cell line BGC-823 and explore role of endoplasmic reticulum stress in apoptosis of gastric cancer cells induced by oxymatrine and elucidate its mechanism. Methods The gastric cancer BGC-823 cells at the logarithmic phase were divided into a control group, oxymatrine alone group (oxymatrine at 10, 30, 60 and 90 μmol/L concentrations), and combination group (oxymatrine at various concentrations combined with 2 μmol/L endoplasmic reticulum stress inhibitor salubrinal). The MTT assay was used to observe the inhibitory effect of the oxymatrine on the growth of BGC-823 cells. The flow cytometry was used to analyze the apoptosis and cell cycle. The Western blot and RT-PCR methods were used to detect the expressions of GRP78/Bip and the caspase-12 protein and gene, respectively. Results ① Compared with the control group, the oxymatrine could significantly inhibit the growth of gastric cancer BGC-823 cells in a concentration-time dependent manner (P<0.05) and its IC50 (48 h) value was (59.5±0.5) μmol/L. The inhibitory effect of the combination group of 30, 60, and 90 μmol/L oxymatrine was significantly weakened as compared with the the corresponding oxymatrine alone group (P<0.05). ② The oxymatrine could significantly induce the apoptosis and arrest the G2/M phase in the gastric cancer BGC-823 cells in a concentration-dependent manner (P<0.05). The combination group of 60 μmol/L oxymatrine could significantly increased the apoptosis rate and the number of cells in the G2/M phase in the gastric cancer BGC-823 cells after treating 48 h (P<0.05). ③ The protein and gene levels of GRP78/Bip and caspase-12 showed a significant increase with the increase of oxymatrine concentrations in the oxymatrine alone group as compared with the control group (P<0.05), which in the combination group of 60 μmol/L and 90 μmol/L oxymatrines were significantly decreased as compared with the corresponding oxymatrine alone group (P<0.05). Conclusion Oxymatrine can inhibit growth of human gastric cancer cell line BGC-823, which may be related to caspase-12-dependent induction of apoptosis and up-regulation of GRP78/Bip expression, which needs further experimental verification.

关键词: 氧化苦参碱; 胃肿瘤; 凋亡; 内质网应激

Key words: oxymatrine; gastric neoplasm; apoptosis; endoplasmic reticulum stress

登录后 ,请手动点击刷新查看全文内容。 没有账号,
登录后 ,请手动点击刷新查看图表内容。 没有账号,
1. Zhou YJ, Guo YJ, Yang XL, et al. Anti-cervical cancer role of matrine, oxymatrine and sophora flavescens alkaloid gels and its mechanism. J Cancer, 2018, 9(8): 1357-1364.
2. 朱栋良, 尹小平, 王芳元. 异鼠李素对胃癌细胞增殖及凋亡的影响. 中国普外基础与临床杂志, 2018, 25(3): 350-353.
3. 王建, 安华松, 宋正霞, 等. 曲妥珠单抗对SW-620人结肠癌细胞增殖凋亡的影响及其与奥沙利铂协同作用的研究. 中国普外基础与临床杂志, 2018, 25(12): 1433-1438.
4. Zhang B, Wang X, Li Y, et al. Matrine is identified as a novel macropinocytosis inducer by a network target approach. Front Pharmacol, 2018, 9: 10.
5. Yin W, Han J, Zhang Z, et al. Aloperine protects mice against bleomycin-induced pulmonary fibrosis by attenuating fibroblast proliferation and differentiation. Sci Rep, 2018, 8(1): 6265.
6. Yin LL, Wen XM, Lai QH, et al. Lenalidomide improvement of cisplatin antitumor efficacy on triple-negative breast cancer cells. Oncol Lett, 2018, 15(5): 6469-6474.
7. Xue JX, Zhu ZY, Bian WH, et al. The traditional Chinese medicine Kangai injection as an adjuvant method in combination with chemotherapy for the treatment of breast cancer in Chinese patients: A meta-analysis. Evid Based Complement Alternat Med, 2018, 2018: 6305645.
8. Xu XX, Bi JP, Ping L, et al. A network pharmacology approach to determine the synergetic mechanisms of herb couple for treating rheumatic arthritis. Drug Des Devel Ther, 2018, 12: 967-979.
9. Meng LQ, Wang Y, Luo YH, et al. Quinalizarin induces apoptosis through reactive oxygen species (ROS)-mediated mitogen-activated protein kinase (MAPK) and Signal transducer and activator of transcription 3(STAT3) signaling pathways in colorectal cancer cells. Med Sci Monit, 2018, 24: 3710-3719.
10. Li W, Yu X, Tan S, et al. Oxymatrine inhibits non-small cell lung cancer via suppression of EGFR signaling pathway. Cancer Med, 2018, 7(1): 208-218.
11. Hou X, Du H, Yang R, et al. The antitumor activity screening of chemical constituents from Camellia nitidissima Chi. Int J Mol Med, 2018, 41(5): 2793-2801.
12. Gu LL, Shen ZL, Li YL, et al. Oxymatrine causes hepatotoxicity by promoting the phosphorylation of JNK and induction of endoplasmic reticulum stress mediated by ROS in LO2 cells. Mol Cells, 2018, 41(5): 401-412.
13. Dai JP, Wang QW, Su Y, et al. Oxymatrine inhibits influenza A virus replication and inflammation via TLR4, p38 MAPK and NF-κB pathways. Int J Mol Sci, 2018, 19(4): pii: E965.
14. Hu Z, Qin F, Gao S, et al. Paeoniflorin exerts protective effect on radiation-induced hepatic fibrosis in rats via TGF-β1/Smads signaling pathway. Am J Transl Res, 2018, 10(3): 1012-1021.
15. Aghvami M, Ebrahimi F, Zarei MH, et al. Matrine induction of ROS mediated apoptosis in human ALL B-lymphocytes via mitochondrial targeting. Asian Pac J Cancer Prev, 2018, 19(2): 555-560.
16. Zhu C, Li X, Zhang B, et al. Quantitative analysis of multi-components by single marker-a rational method for the internal quality of Chinese herbal medicine. Integr Med Res, 2017, 6(1): 1-11.
17. Zhang Y, Liang Y, He C. Anticancer activities and mechanisms of heat-clearing and detoxicating traditional Chinese herbal medicine. Chin Med, 2017, 12: 20.