药学学报, 2021, 56(8): 2223-2229
引用本文:
叶汉林, 乔淦, 王琳琳, 程丽, 林秀坤*. 原阿片碱通过线粒体凋亡途径抑制肝细胞癌生长[J]. 药学学报, 2021, 56(8): 2223-2229.
YE Han-lin, QIAO Gan, WANG Lin-lin, CHENG Li, LIN Xiu-kun*. Protopine inhibits the growth of hepatocellular carcinoma through a mitochondrially mediated apoptosis pathway[J]. Acta Pharmaceutica Sinica, 2021, 56(8): 2223-2229.

原阿片碱通过线粒体凋亡途径抑制肝细胞癌生长
叶汉林, 乔淦, 王琳琳, 程丽, 林秀坤*
西南医科大学药学院, 四川 泸州 646000
摘要:
全球范围内超过一半的肝癌病例发生在我国,亟待发现新的抗肝癌药物。已有报道表明,原阿片碱具有较好的抗肿瘤活性,但其作用机制尚不清楚。本研究采用CCK-8实验证实原阿片碱对肝癌细胞具有显著的抑制作用,流式细胞术和JC-1染色实验表明原阿片碱能够降低肝癌细胞线粒体膜电位、诱导肝癌凋亡,Western blot分析表明原阿片碱能够激活线粒体凋亡通路,上调Bax、细胞色素C (cytochrome C,Cyto-C)、剪切的半胱天冬氨酸蛋白酶-9(cleaved-caspase-9)、剪切的半胱天冬氨酸蛋白酶-3(cleaved-caspase-3)、剪切的DNA修复酶(cleaved-poly ADPribose polymerase,cleaved-PARP)和凋亡酶激活因子-1(apoptotic protease activating factor-1,Apaf-1)的表达,下调Bcl-2的表达。人肝癌裸鼠移植性肿瘤实验证实,50 mg·kg-1原阿片碱腹腔注射给药,抑瘤率可达到72.46%,并对小鼠体重没有明显影响。动物实验严格按照西南医科大学动物伦理委员会的规定执行。本研究提示,原阿片碱有潜力开发为治疗肝细胞癌的药物前体。
关键词:    肝细胞癌      原阿片碱      抗肿瘤活性      线粒体凋亡     
Protopine inhibits the growth of hepatocellular carcinoma through a mitochondrially mediated apoptosis pathway
YE Han-lin, QIAO Gan, WANG Lin-lin, CHENG Li, LIN Xiu-kun*
School of Pharmacy, Southwest Medical University, Luzhou 646000, China
Abstract:
Hepatocellular carcinoma (HCC) is a serious threat for human health, the incidence of HCC in China accounts for more than 50% worldwide. There is an urgent need to develop novel anticancer agents for the treatment of HCC patients. Here we characterized the inhibitory effect and the molecular mechanism of protopine on HCC cancer cells. The results of a CCK-8 assay indicated that protopine displays anticancer activities on HCC cells. Flow cytometry and JC-1 staining confirmed that treatment with protopine decreased the mitochondrial membrane potential and induced apoptosis in HCC cells. Western blot analysis showed that protopine was able to increase protein expression in the mitochondrial apoptotic pathway; the level of cytochrome C, apoptotic protease activating factor-1 (Apaf-1), Bax, cleaved-poly ADP-ribose polymerase (cleaved-PARP), cleaved-caspase-3, and cleaved-caspase-9 were increased while the expression of Bcl-2 was suppressed significantly. An in vivo study revealed that protopine significantly suppressed the growth of tumors in nude mice bearing HepG2 cells. Administration of protopine intraperitoneally at a concentration of 50 mg·kg-1 inhibited tumor growth by 72.46%. Animal experiments were carried out according to the Regulation of the Animal Ethics Committee of Southwest Medical University. This study provides preliminary evidence that there is potential to develop protopine as a lead compound for the treatment of HCC.
Key words:    hepatocellular carcinoma    protopine    anticancer activity    mitochondrial apoptosis   
收稿日期: 2021-03-07
DOI: 10.16438/j.0513-4870.2021-0319
基金项目: 国家自然科学基金资助项目(82773776).
通讯作者: 林秀坤,Tel:13522138991,E-mail:xiukunlin@126.com
Email: xiukunlin@126.com
相关功能
PDF(795KB) Free
打印本文
0
作者相关文章
叶汉林  在本刊中的所有文章
乔淦  在本刊中的所有文章
王琳琳  在本刊中的所有文章
程丽  在本刊中的所有文章
林秀坤*  在本刊中的所有文章

参考文献:
[1] Torre LA, Bray F, Siegel RL, et al. Global cancer statistics, 2012[J]. CA Cancer J Clin, 2015, 65:87-108.
[2] Ferlay J, Colombet M, Soerjomataram I, et al. Estimating the global cancer incidence and mortality in 2018:GLOBOCAN sources and methods[J]. Int J Cancer, 2019, 144:1941-1953.
[3] Chen W, Zheng R, Baade PD, et al. Cancer statistics in China, 2015[J]. CA Cancer J Clin, 2016, 66:115-132.
[4] Fu J, Wang H. Precision diagnosis and treatment of liver cancer in China[J]. Cancer Lett, 2017, 412:283-288.
[5] Faivre S, Rimassa L, Finn RS. Molecular therapies for HCC:looking outside the box[J]. J Hepatol, 2020, 72:342-352.
[6] Villanueva A, Llovet JM. Second-line therapies in hepatocellular carcinoma:emergence of resistance to sorafenib[J]. Clin Cancer Res, 2012, 18:1824-1826.
[7] Sung DK, Kim YH, Pan CH, et al. Protopine reduces the inflammatory activity of lipopolysaccharide-stimulated murine macrophages[J]. BMB Rep, 2012, 45:108-113.
[8] Orhan I, Ozçelik B, Karaoğlu T, et al. Antiviral and antimicrobial profiles of selected isoquinoline alkaloids from Fumaria and Corydalis species[J]. Z Naturforsch C J Biosci, 2007, 62:19-26.
[9] Rathi A, Srivastava AK, Shirwaikar A, et al. Hepatoprotective potential of Fumaria indica Pugsley whole plant extracts, fractions and an isolated alkaloid protopine[J]. Phytomedicine, 2008, 15:470-477.
[10] He K, Gao J. Protopine inhibits heterotypic cell adhesion in MDAMB-231 cells through down-regulation of multi-adhesive factors[J]. Afr J Tradit Complement Altern Med, 2014, 11:415-424.
[11] Chen CH, Liao CH, Chang YL, et al. Protopine, a novel microtubule-stabilizing agent, causes mitotic arrest and apoptotic cell death in human hormone-refractory prostate cancer cell lines[J]. Cancer Lett, 2012, 315:1-11.
[12] Son YL, An YJ, Jung J, et al. Protopine isolated from Nandina domestica induces apoptosis and autophagy in colon cancer cells by stabilizing p53[J]. Phytother Res, 2019, 33:1689-1696.
[13] Zhang X, Meng LH. Progress in molecularly targeted anti-tumor drugs derived from natural products or their derivatives[J]. Acta Pharm Sin (药学学报), 2020, 55:2491-2500.
[14] Yang CP, Horwitz S. Taxol:the first microtubule stabilizing agent[J]. Int J Mol Sci, 2017, 18:1733.
[15] Martino E, Della VS, Terribile E, et al. The long story of camptothecin:from traditional medicine to drugs[J]. Bioorg Med Chem Lett, 2016, 27:701-707.
[16] Shah NN, Merchant MS, Cole DE, et al. Vincristine sulfate liposomes injection (VSLI, Marqibo):results from a phase I study in children, adolescents, and young adults with refractory solid tumors or leukemias[J]. Pediatr Blood Cancer, 2016, 63:997-1005.
[17] Hermeking H. The miR-34 family in cancer and apoptosis[J]. Cell Death Differ, 2010, 17:193-199.
[18] Wiraswati HL, Hangen E, Sanz AB, et al. Apoptosis inducing factor (AIF) mediates lethal redox stress induced by menadione[J]. Oncotarget, 2016, 7:76496-76507.
[19] Weaver BA. How taxol/paclitaxel kills cancer cells[J]. Mol Biol Cell, 2014, 25:2677-2681.
[20] Wyatt MD, Wilson DM. Participation of DNA repair in the response to 5-fluorouracil[J]. Cell Mol Life Sci, 2009, 66:788-799.
[21] Liew ST, Yang LX. Design, synthesis and development of novel camptothecin drugs[J]. Curr Pharm Des, 2008, 14:1078-1097.