白花蛇舌草对人胃癌细胞MNK-45线粒体膜电位及凋亡相关基因表达的影响*

doi:10.12047/j.cjap.5956.2020.038

中国应用生理学杂志 ›› 2020, Vol. 36 ›› Issue (2): 171-175.doi: 10.12047/j.cjap.5956.2020.038

白花蛇舌草对人胃癌细胞MNK-45线粒体膜电位及凋亡相关基因表达的影响^*

杜洋¹, 邵淑丽^1,2△, 焦凯贺¹, 刘祥露¹, 冯元¹, 张伟伟^1,2

1.齐齐哈尔大学生命科学与农林学院, 2.抗性基因工程与寒地生物多样性保护黑龙江省重点实验室, 黑龙江齐齐哈尔 161006

出版日期:2020-03-28 发布日期:2020-07-31
通讯作者: ^△Tel: 13836267458; E-mail: shshl32@163.com
基金资助:
^*黑龙江省自然科学基金(C201241);黑龙江省教育厅基本业务专项重点项目(135109104);黑龙江省高教强省优势特色学科—玉米“粮头食尾”重点专项(LTSW201737);黑龙江省省属高等学校基本科研业务费科研项目(植物性食品加工技术特色学科专项)(YSTSXK201809)

Effects of hedyotis diffusa on mitochondrial membrane potential and expressions of apoptosis-related genes in human gastric cancer cell line MNK-45

DU Yang¹, SHAO Shu-li^1,2△, JIAO Kai-he¹, LIU Xiang-lu¹, FENG Yuan¹, ZHANG Wei-wei^1,2

1. College of Life Sciences, Agriculture and Forestry; 2. Heilongjiang Provincial Key Laboratory of Resistance Gene Engineering and Protection of Biodiversity in Cold Areas, Qiqihar University, Qiqihar 161006, China

Online:2020-03-28 Published:2020-07-31

摘要/Abstract

摘要： 目的: 探讨白花蛇舌草(注射液)对人胃癌细胞MNK-45线粒体膜电位及凋亡相关基因表达的影响。方法: 将人胃癌细胞MNK-45分成4组,每组设置3个复孔,对照组为未加入白花蛇舌草的MNK-45细胞,3组实验组分别加入终浓度为20、30、40 μg/ml白花蛇舌草的MNK-45细胞,各组在5％的CO₂培养箱中孵育48 h后,利用激光共聚焦显微镜下观察细胞形态变化,流式细胞术检测线粒体膜电位,qRT-PCR检测Cytochrome C (Cyt c)、caspase3和caspase9基因的表达变化,Western blot检测Cytochrome C (Cyt c)、caspase3和caspase9蛋白的表达变化。结果: 与对照组相比,终浓度为20、30、40 μg/ml的各白花蛇舌草处理组,其MNK-45细胞的线粒体膜电位均明显降低(P＜0.01),Cyt c、caspase3和caspase9的基因表达均明显上调(P＜0.01)、蛋白表达也均显著升高(P＜0.05或 P＜0.01),40 μg/ml的白花蛇舌草处理组的表现最佳。结论: 在终浓度为20~40 μg/ml的范围内,白花蛇舌草能够降低人胃癌MNK-45细胞线粒体膜电位, 诱导细胞凋亡,并可上调Cyt c、caspase3和caspase9基因表达。

关键词: 白花蛇舌草, 线粒体膜电位, 人胃癌MNK-45细胞, 细胞色素C, caspase3, caspase9

Abstract: Objective: To investigate the effects of hedyotis diffusa (injection) on mitochondrial membrane potential and expressions of apoptosis-related genes in human gastric cancer cell line MNK-45 cells. Methods: The human gastric cancer MNK-45 cells were divided into 4 groups, each group was set with 3 replicates. The control group was MNK-45 cells without added hedyotis diffusa; the 3 groups of experimental groups were treated with hedyotis diffusa at final concentrations of 20 , 30, 40 μg / ml respectively; each group was incubated for 48 h in a 5% carbon dioxide incubator, and the morphological changes of the cells were observed under a laser confocal microscope. Mitochondrial membrane potential was detected by flow cytometry. The expressions of Cytochrome C (Cyt c), caspase3 and caspase9 genes and proteins were detected by qRT-PCR and Western blot respectively. Results: Compared with the control group, the mitochondrial membrane potentials of MNK-45 cells were significantly reduced in the hedyotis diffusa treated groups at final concentrations of 20, 30, and 40 μg / ml (P＜0. 01). The gene expressions of Cyt c, caspase3, and caspase9 were significantly up-regulated (P＜0. 01) and their protein expressions were also significantly increased (P＜0. 05 or P＜0. 01). The 40 μg / ml hedyotis diffusa treatment group performed best. Conclusion: In the final concentration range of 20 ~ 40 μg / ml, hedyotis diffusa can reduce human gastric cancer MNK-45 cells mitochondrial membrane potential, induce apoptosis and up-regulate Cyt c, caspase3 and caspase9 gene expressions.

Key words: hedyotis diffusa, mitochondrial membrane potential, human gastric cancer MNK-45 cells, Cyt c, caspase3, caspase9

中图分类号:

Q255

杜洋, 邵淑丽, 焦凯贺, 刘祥露, 冯元, 张伟伟. 白花蛇舌草对人胃癌细胞MNK-45线粒体膜电位及凋亡相关基因表达的影响^*[J]. 中国应用生理学杂志, 2020, 36(2): 171-175.

DU Yang, SHAO Shu-li, JIAO Kai-he, LIU Xiang-lu, FENG Yuan, ZHANG Wei-wei. Effects of hedyotis diffusa on mitochondrial membrane potential and expressions of apoptosis-related genes in human gastric cancer cell line MNK-45[J]. CJAP, 2020, 36(2): 171-175.

参考文献

[1] 耿龙龙, 任鹏, 李延海, 等. 胃癌治疗现状及新进展[J]. 医学理论与实践, 2017, 30(12): 1744-1746.
[2] 朱正伦, 燕敏. 早期胃癌的外科治疗策略[J]. 腹腔镜外科杂志, 2017, 22(1): 1-6.
[3] D'Angelica M, Gonen M, Brennan MF, et al. Patterns of initial recurrence in completely resected gastric adenocarcinoma[J]. Ann Surg, 2004, 240(5) : 808-816.
[4] 叶昀旖. 益气健脾化积方对胃癌化疗患者免疫调控作用的临床研究[D]. 南京: 南京中医药大学, 2017.
[5] 欧水平, 王森, 陈灵, 等. 白花蛇舌草-半枝莲药对配伍研究分析[J]. 中国中医基础医学杂志, 2015, 21(2): 215-217.
[6] Hu E, Wang D, Chen J, et al. Novel cyclotides from Hedyotis diffusa induce apoptosis and inhibit proliferation and migration of prostate cancer cells[J]. Int J Clin Exp Med, 2015, 8 (3) : 4059-4065.
[7] Langheinrich MC, Schellerer V, Perrakis A, et al. Molecular mechanisms of lymphatic metastasis in solid tumors of the gastrointestinal tract[J]. Int J Clin Exppathol, 2012, 5(7) : 614-623.
[8] Han Z, Hong L, Han Y, et al. Phospho Akt mediates multidrug resistance of gastric cancer cells through regulation of P-gp, Bcl-2 and Bax[J]. J Exp Clin Cancer Res, 2007, 26(2): 261-268.
[9] Khalil DN, Smith EL, Brentjens RJ, et al. The future of cancer treatment: immunomodulation, CARs and combination immunotherapy[J]. Nat Revi Clin Oncol, 2016, 13(5) : 273-273.
[10]Kim DH, Lee HJ, Oh YJ, et al. Iridoid glycosides isolated from Oldenlandia diffusa inhibit LDL-oxidation[J]. Arch Pharm Res, 2005, 28(10) : 1156-1160.
[11]曾珠, 何成诗, 孙剑峰, 等. 白花蛇舌草对A549肺癌干细胞增殖、凋亡及细胞周期的影响[J]. 重庆医学, 2019, 48(13): 2202-2205.
[12]Li H, Lai ZJ, Yang H, et al. Hedyotis diffusa Willd. inhibits VEGF-C-mediated lymphangiogenesis in colorectal cancer via multiple signaling pathways[J]. Oncology reports, 2019, 42(3) : 1225-1236.
[13]Cha Z, Cheng JZ, Xiang H, et al. Celastrol enhances TRAIL-induced apoptosis in human glioblastoma via the death receptor pathway[J]. Cancer Chemoth Pharm, 2019, 84 (4) : 719-728.
[14]Wang H, Liu H, Zheng ZM, et al. Role of death receptor, mitochondrial and endoplasmic reticulum pathways in different stages of degenerative human lumbar disc[J]. Apoptosis, 2011, 16(10): 990-1003.
[15]李欲轲, 熊孟连, 徐僡, 等. 内质网应激与凋亡研究进展[J]. 分子植物育种, 2018,16(23): 7856-7862.
[16]Zhang J, Yi T, Liu J, et al. Quercetin induces apoptosis via the mitochondrial pathway in KB and KBv200 cells[J]. J Agric Food Chem, 2013, 61(9): 2188-2195.
[17]李凤岩, 陈亚光, 林大勇, 等. 甘草次酸对甲状腺癌细胞SW579凋亡的影响及其机制[J]. 中国应用生理学杂志, 2019, 35(5): 464-467.
[18]洪凯, 蒋盼若, 柯瑞君, 等. 2-12烷基-6-甲氧基环己基-2,5-二烯-1,4-二酮(DMDD)抗弥漫大B淋巴瘤的作用及机制[J]. 中国应用生理学杂志, 2019, 35(4): 312-316.
[19]Dai ZJ, Wang XJ, Li ZF, et al. Scutellaria barbate extract induces apoptosis of hepatoma H22 cells via the mitochondrial pathway involving caspase-3.[J]. World J Gastroenterol, 2008, 14(48): 7321-7328.
[20]蒋显. 线粒体释放细胞凋亡因子的机理研究[D]. 北京: 北京协和医学院, 2014.

白花蛇舌草对人胃癌细胞MNK-45线粒体膜电位及凋亡相关基因表达的影响^*

Effects of hedyotis diffusa on mitochondrial membrane potential and expressions of apoptosis-related genes in human gastric cancer cell line MNK-45

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