为了深入明晰酚酸类物质阿魏酸的具体功能机理,以其对CT-26细胞自噬、凋亡、细胞增殖和迁移为评价手段,考察阿魏酸对细胞生理活性的影响,阐明酚酸类物质阿魏酸具体作用机理,为中草药中有效成分阿魏酸的具体功效提供实验数据和理论依据。研究中采用不同浓度梯度的阿魏酸处理CT-26细胞,考察阿魏酸对CT-26细胞生理活性的影响。在一定浓度范围内,阿魏酸能够抑制CT-26细胞的增殖、提高细胞自噬水平,同时在一定程度上抑制CT-26细胞的迁移能力。进一步的研究表明,阿魏酸通过促进凋亡通路上游的caspase3蛋白剪切为Cleaved caspase3,进而调控凋亡关键蛋白Bcl-2和BCL2-Associated X蛋白(Bax)的比值,最终促进了结肠癌细胞凋亡,以此保护细胞的正常生理活动。阿魏酸可通过促进细胞自噬并调控凋亡通路Bcl-2来抑制细胞活性,进而促进细胞凋亡。研究结果为明确中草药中有效成分阿魏酸的生物活性物质作用提供了实验数据。
This study aimed to investigate the specific mechanism of action of the phenolic acid compound ferulic acid. The impact of ferulic acid on cell physiological activity was investigated by using its effects on CT-26 cell autophagy, apoptosis, cell proliferation, and migration as evaluation methods. This study provides experimental data and theoretical basis for the specific efficacy of the active ingredient ferulic acid in traditional Chinese medicine.Colon cancer cell line CT-26 was treated with different concentrations of ferulic acid (FA) to study whether ferulic acid could inhibit CT-26 cell.Within a certain concentration range, ferulic acid could inhibit the proliferation of CT-26 cells, slightly increase the level of autophagy, and inhibit the migration of CT-26 cells to a certain extent.In addition, ferulic acid could promote the cleavage of caspase-3 protein into cleaved caspase-3 protein, and then regulated the proportion of apoptosis Bcl-2 and Bax, so as to promote the apoptosis of colon cancer cells and protect the normal physiological activities of the body.Ferulic acid may inhibit tumor activity and promote cancer cell apoptosis by promoting autophagy and regulating apoptosis pathway bcl-2.The results provided experimental evidence for the bioactive substance of ferulic acid in Chinese herbal medicine.
[1] ZHENG Y H, YOU X R, CHEN L, et al.Biotherapeutic nanoparticles of poly (ferulic acid) delivering doxorubicin for cancer therapy[J].Journal of Biomedical Nanotechnology, 2019, 15(8):1734-1743.
[2] PEARLSTEIN E, AMBROGIO C, KARPATKIN S.Effect of antiplatelet antibody on the development of pulmonary metastases following injection of CT26 colon adenocarcinoma, Lewis lung carcinoma, and B16 amelanotic melanoma tumor cells into mice[J].Cancer Research, 1984, 44(9):3884-3887.
[3] HUANG H Z, HUANG Y X, CHEN Y A, et al.A novel immunochemotherapy based on targeting of cyclooxygenase and induction of immunogenic cell death[J].Biomaterials, 2021, 270(3):120708.
[4] SHOKRZADEH M, AZADBAKHT M, AHANGAR N, et al.Cytotoxicity of hydro-alcoholic extracts of Cucurbita pepo and Solanum nigrum on HepG2 and CT26 cancer cell lines[J].Pharmacognosy Magazine, 2010, 6(23):176-179.
[5] LEE K W, CHOI B, KIM Y M, et al.Major histocompatibilty complex-restricted adaptive immune responses to CT26 colon cancer cell line in mixed allogeneic Chimera[J].Transplantation Proceedings, 2017, 49(5):1153-1159.
[6] KERR J F R, WYLLIE A H, CURRIE A R.Apoptosis:A basic biological phenomenon with wide-ranging implications in tissue kinetics[J].British Journal of Cancer, 1972, 26(4):239-257.
[7] FESIK S W.Promoting apoptosis as a strategy for cancer drug discovery[J].Nature Reviews Cancer, 2005, 5(11):876-885.
[8] MATHEW R, KARANTZA-WADSWORTH V, WHITE E.Role of autophagy in cancer[J].Nature Reviews Cancer, 2007, 7(12):961-967.
[9] YUE Z Y, JIN S K, YANG C, et al.Beclin 1, an autophagy gene essential for early embryonic development, is a haploinsufficient tumor suppressor[J].Proceedings of the National Academy of Sciences of the United States of America, 2003, 100(25):15077-15082.
[10] MAIURI M M C, ZALCKVAR E, KIMCHI A, et al.Self-eating and self-killing:Crosstalk between autophagy and apoptosis[J].Nature Reviews Molecular Cell Biology, 2007, 8(9):741-752.
[11] PATTINGRE S, TASSA A, QU X P, et al.Bcl-2 antiapoptotic proteins inhibit beclin 1-dependent autophagy[J].Cell, 2005, 122(6):927-939.
[12] LUM J J, BAUER D E, KONG M, et al.Growth factor regulation of autophagy and cell survival in the absence of apoptosis[J].Cell, 2005, 120(2):237-248.
[13] BOYA P, GONZÁLEZ-POLO R A, CASARES N, et al.Inhibition of macroautophagy triggers apoptosis[J].Molecular and Cellular Biology, 2005, 25(3):1025-1040.
[14] GOZUACIK D, KIMCHI A.Autophagy as a cell death and tumor suppressor mechanism[J].Oncogene, 2004, 23(16):2891-2906.
[15] DEGENHARDT K, MATHEW R, BEAUDOIN B, et al.Autophagy promotes tumor cell survival and restricts necrosis, inflammation, and tumorigenesis[J].Cancer Cell, 2006, 10(1):51-64.
[16] SACILOTO B, NICOLETTI N I F, PELETTI-FIGUEIRÓ M, et al.Crosstalk between autophagy and apoptosis in intervertebral disc degeneration[J].Journal of Bioscience and Medicines, 2021, 9(12):15-29.
[17] MOHAMED S M, MOHAMMED D S, ELHALIEM N G A, et al.Mangosteen can improve steatohepatitis through modulating inflammatory and autophagy/apoptosis cell injury:An animal model study[J].Cytology and Genetics, 2021, 55(5):480-490.
[18] KAWAGUCHI Y, KOVACS J J, MCLAURIN A, et al.The deacetylase HDAC6 regulates aggresome formation and cell viability in response to misfolded protein stress[J].Cell, 2003, 115(6):727-738.
[19] RODRIGUEZ L G, WU X Y, GUAN J L.Wound-healing assay[J].Methods in Molecular Biology, 2005, 294(3):23-29.
[20] FAHRIOĞLU U, DODURGA Y, ELMAS L, et al.Ferulic acid decreases cell viability and colony formation while inhibiting migration of MIA PaCa-2 human pancreatic cancer cells in vitro[J].Gene, 2016, 576(1):476-482.
[21] KOLLMAR O, RUPERTUS K, SCHEUER C, et al.CXCR4 and CXCR7 regulate angiogenesis and CT26.WT tumor growth independent from SDF-1[J].International Journal of Cancer, 2010, 126(6):1302-1315.
[22] KORSMEYER S J, SHUTTER J R, VEIS D J, et al.Bcl-2/Bax:A rheostat that regulates an anti-oxidant pathway and cell death[J].Seminars in Cancer Biology, 1993, 4(6):327-332.
[23] YANG J, LIU X, BHALLA K, ET AL.Prevention of apoptosis by Bcl-2:Release of cytochrome c from mitochondria blocked[J].Science, 1997, 275(5303):1129-1132.
[24] GOWN A M, WILLINGHAM M C.Improved detection of apoptotic cells in archival paraffin sections:Immunohistochemistry using antibodies to cleaved caspase 3[J].The Journal of Histochemistry and Cytochemistry:Official Journal of the Histochemistry Society, 2002, 50(4):449-454.
[25] PANKIV S, CLAUSEN T H, LAMARK T, et al.p62/SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy[J].Journal of Biological Chemistry, 2007, 282(33):24131-24145.
[26] WANG J, HE J, SU F, et al.Repression of ATR pathway by miR-185 enhances radiation-induced apoptosis and proliferation inhibition[J].Cell Death & Disease, 2013, 4(6):e699.
[27] ZHANG S S, NIE S P, HUANG D F, et al.Immunomodulatory effect of Ganoderma atrum polysaccharide on CT26 tumor-bearing mice[J].Food Chemistry, 2013, 136(3-4):1213-1219.
[28] ZHANG K, ZHOU X T, WANG J Q, et al.Dendrobium officinale polysaccharide triggers mitochondrial disorder to induce colon cancer cell death via ROS-AMPK-autophagy pathway[J].Carbohydrate Polymers, 2021, 264:118018.
