6-姜烯酚诱导SW480凋亡及对APC基因表达的影响
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摘要
本文通过体外细胞实验观察研究6-姜烯酚诱导结直肠癌细胞凋亡及与APC(肿瘤抑制基因)表达的关系,以探讨6-姜烯酚抑制结直肠肿瘤的可能机制。首先通过体外细胞实验应用不同浓度6-姜烯酚(0、5、10、15和20μM)分别对SW480进行干预诱导24 h,再利用高倍荧光显微镜观察细胞数目、形态变化,CCK8法测定细胞抑制率,Annexin V-FITC/PI流式细胞术检测细胞凋亡率及细胞凋亡周期,最后用Western-blot检测分析APC蛋白表达变化。结果显示,与对照组相比,6-姜烯酚可致使SW480细胞增殖周期阻断在G2/M期,并呈浓度依赖性地促进SW480凋亡,进而抑制细胞增殖,而且增强了APC表达(p<0.05)。因此,可以得出6-姜烯酚可诱导SW480凋亡从而抑制细胞增殖,并且可能与激活APC相关,随着这一研究的不断深入,6-姜烯酚作为临床潜在的抗肿瘤辅助药物,其抑制肿瘤细胞增殖将得到更深层次分子机制的支持。
The relationship between the apoptosis of human colorectal cancer cells(induced by 6-shogaol) and the expression of APC was investigated to study the possible mechanism of 6-shogaol inhibiting colorectal cancer in this study. SW480 was induced by different concentrations of 6-shogoal(0, 5, 10, 15 and 20 μM) in vitro for 24 h. Then the number and morphology of cells were observed by high-frequency fluorescence microscopy, the cell inhibition rate was determined by CCK8 assay, and the apoptosis rate and apoptosis cycle were detected by Annexin V-FITC/PI flow cytometry. In addition, the expression of APC protein was analyzed by western-blot. The results showed that compared with the control group, 6-shogaol could block the proliferation cycle of SW480 in G2/M phase and promote the apoptosis of SW480 in a concentration dependent manner, thereby inhibiting the cell proliferation and enhancing the expression of APC(p<0.05). Consequently, 6-shogaol could induce SW480 apoptosis and inhibit cell proliferation, and might be associated with the activation of APC. With the deepening of this study, the inhibition of tumor cell proliferation induced by 6-shogaol(used as a potential antitumor drug) would get more support of deeper molecular mechanism.
The relationship between the apoptosis of human colorectal cancer cells(induced by 6-shogaol) and the expression of APC was investigated to study the possible mechanism of 6-shogaol inhibiting colorectal cancer in this study. SW480 was induced by different concentrations of 6-shogoal(0, 5, 10, 15 and 20 μM) in vitro for 24 h. Then the number and morphology of cells were observed by high-frequency fluorescence microscopy, the cell inhibition rate was determined by CCK8 assay, and the apoptosis rate and apoptosis cycle were detected by Annexin V-FITC/PI flow cytometry. In addition, the expression of APC protein was analyzed by western-blot. The results showed that compared with the control group, 6-shogaol could block the proliferation cycle of SW480 in G2/M phase and promote the apoptosis of SW480 in a concentration dependent manner, thereby inhibiting the cell proliferation and enhancing the expression of APC(p<0.05). Consequently, 6-shogaol could induce SW480 apoptosis and inhibit cell proliferation, and might be associated with the activation of APC. With the deepening of this study, the inhibition of tumor cell proliferation induced by 6-shogaol(used as a potential antitumor drug) would get more support of deeper molecular mechanism.
引文
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[17]Liu S,Tackmann N R,Yang J,et al.Disruption of the RP-MDM2-p53 pathway accelerates APC loss-induced colorectal tumorigenesis[J].Oncogene,2017,36(10):1374
[18]Gausachs M,Borras E,Chang K,et al.Mutational Heterogeneity in APC and KRAS Arises at the Crypt level and Leads to Polyclonality in Early Colorectal Tumorigenesis[J].Clinical cancer research:an official journal of the American Association for Cancer Research,2017
[19]邱淑敏,陈涛,刘美玲,等.中药诱导肿瘤细胞凋亡机制研究进展[J].动物医学进展,2015,1:83-86QIU Shu-min,CHEN Tao,LIU Mei-ling,et al.Advances in research on mechanism of chinese medicine inducing apoptosis of tumor cells[J].Advances in Animal Medicine,2015,1:83-86
[20]苗久旺,张钦德.姜黄素诱导肿瘤细胞凋亡的信号途径研究进展[J].陕西中医,2014,12:1692-1694MIAO Jiu-wang,ZHANG Qin-de.Progress in signal pathway of curcumin inducing apoptosis of tumor cells[J].Chinese Medical Science of Shaanxi,2014,12:1692-1694
[21]Hung J Y,Hsu Y L,Li C T,et al.6-Shogaol,an active constituent of dietary ginger,induces autophagy by inhibiting the AKT/m TOR pathway in human non-small cell lung cancer A549 cells[J].J.Agric.Food Chem.,2009,57(20):9809-9816
[22]Tan B S,Kang O,Mai C W,et al.6-Shogaol inhibits breast and colon cancer cell proliferation through activation of peroxisomal proliferator activated receptor gamma(PPAR gamma)[J].Cancer Lett.,2013,336:127-139
[23]Park S J,Lee M Y,Son B S,et al.TBK1-targeted suppression of TRIF-dependent signaling pathway of Toll-like receptors by 6-shogaol,an active component of ginger[J].Biosci.,Biotechnol.,Biochem.,2009,73(7):1474-1478
[2]杨奕夫.结直肠病死率和并发症率的生理学及手术严重性评分在结直肠癌诊疗中的研究进展[J].中国微创外科杂志,2017,17(5):458-460YANG Yi-fu.Advances in physiology and severity of colorectal cancer mortality and complication rates in colorectal cancer diagnosis and treatment[J].Chinese Journal of Minimally Invasive Surgery,2017,17(5):458-460
[3]许敬尧.结直肠肿瘤WHO分类新进展以及诊断与鉴别诊断[C]//浙江省病理学术年会,2013XU Jing-yao.New progress in WHO classification and diagnosis and differential diagnosis of colorectal cancer[C]//annual meeting of Pathology Society of Zhejiang,2013
[4]王伟.中药在治疗肿瘤方面的作用及其有效成分分析[J].内蒙古中医药,2017,36(3):109-109WANG Wei.Effect of Chinese herbal medicine on the treatment of tumor and analysis of its effective components[J].Chinese Medicine of Inner Mongolia,2017,36(3):109-109
[5]Wilson R,Haniadka R,Sandhya P,et al.Ginger(Zingiber officinale,Roscoe)the dietary agent in skin care:a review[M].Bioactive Dietary Factors and Plant Extracts in Dermatology,Humana Press,2013
[6]赵文竹,李思慧,宋宝雯,等.生姜多糖类物质的提取及抗氧化活性研究[J].食品安全质量检测学报,2017,8(4):1357-1362ZHAO Wen-zhu,LI Si-hui,SONG Bao-wen,et al.Studies on extraction and antioxidant activities of polysaccharides from ginger[J].Journal of Food Safety and Quality Control,2017,8(4):1357-1362
[7]Chen H,Fu J,Chen H,et al.Ginger compound
[6]-shogaol and its cysteine-conjugated metabolite(M2)activate Nrf2 in colon epithelial cells in vitro and in vivo[J].Chemical Research in Toxicology,2014,27(9):1575-85
[8]Zick S M,Djuric Z,Ruffin M T,et al.Pharmacokinetics of 6-,8-,10-Gingerols and 6-Shogaol and conjugate metabolites in healthy human subjects[J].Cancer epidemiology,biomarkers&prevention:a publication of the American Association for Cancer Research,cosponsored by the American Society of Preventive Oncology,2008,17(8):1930
[9]Kim S O,Kundu J K Shin Y K,Park J H,et al.(6)-Gingerol inhibits COX-2 expression by blocking the activation of p38MAP kinase and NF-kappa B in phorbol ester-stimulated mouse skin[J].Oncogene,2005,24(15):2558-67
[10]Luo X J,Xuan-Rong M O,Zhou L L.The effect of Hsp72 on IL-6,IL-8 expression and activation of NF-κB in synoviocytes of rheumatoid arthritis[J].Chinese Journal of Applied Physiology,2012,28(4):336
[11]Kim S O,Chun K S,Kundu J K,et al.Inhibitory effects of
[6]-gingerol on PMA-induced COX-2 expression and activation of NF-kappa B and p38 MAPK in mouse skin[J].Biofactors,2004,21(4):27-31
[12]Nigam N,Bhui K,Prasad S,et al.
[6]-Gingerol induces reactive oxygen species regulated mitochondrial cell death pathway in human epidermoid carcinoma A431 cells[J].Chemico-Biological Interactions,2009,181(1):77
[13]Chakraborty D,Bishayee K,Ghosh S,et al.
[6]-Gingerol induces caspase 3 dependent apoptosis and autophagy in cancer cells:drug-DNA interaction and expression of certain signal genes in He La cells[J].European Journal of Pharmacology,2012,694(1-3):20-29
[14]杨平,曹杰,张通,等.藤黄酸抑制人结肠癌SW480细胞增殖过程中APC蛋白的表达变化[J].热带医学杂志,2013,13(6):726-728,736YANG Ping,CAO Jie,ZHANG Tong,et al.Gambogic acid inhibits APC protein on proliferation of human colon cancer SW480 cells the expression of[J].Tropical Med.,2013,13(6):726-728736
[15]刘杨,巩阳,林一帆,等.散发性大肠腺瘤性息肉患者粪便中APC基因突变的意义及与中医体质的关系[J].中国中西医结合消化杂志,2017,5:327-331LIU Yang,GONG Yang,LIN Yi-fan,et al.Significance of APC gene mutations in feces of sporadic colorectal adenomatous polyps and its relationship with constitution of Chinese medicine[J].Chinese Journal of Integrative Medicine Digest,2017,5:327-331
[16]Sansregret L,Patterson J O,Dewhurst S,et al.APC/C dysfunction limits excessive cancer chromosomal instability[J].Cancer Discovery,2017,7(2):218
[17]Liu S,Tackmann N R,Yang J,et al.Disruption of the RP-MDM2-p53 pathway accelerates APC loss-induced colorectal tumorigenesis[J].Oncogene,2017,36(10):1374
[18]Gausachs M,Borras E,Chang K,et al.Mutational Heterogeneity in APC and KRAS Arises at the Crypt level and Leads to Polyclonality in Early Colorectal Tumorigenesis[J].Clinical cancer research:an official journal of the American Association for Cancer Research,2017
[19]邱淑敏,陈涛,刘美玲,等.中药诱导肿瘤细胞凋亡机制研究进展[J].动物医学进展,2015,1:83-86QIU Shu-min,CHEN Tao,LIU Mei-ling,et al.Advances in research on mechanism of chinese medicine inducing apoptosis of tumor cells[J].Advances in Animal Medicine,2015,1:83-86
[20]苗久旺,张钦德.姜黄素诱导肿瘤细胞凋亡的信号途径研究进展[J].陕西中医,2014,12:1692-1694MIAO Jiu-wang,ZHANG Qin-de.Progress in signal pathway of curcumin inducing apoptosis of tumor cells[J].Chinese Medical Science of Shaanxi,2014,12:1692-1694
[21]Hung J Y,Hsu Y L,Li C T,et al.6-Shogaol,an active constituent of dietary ginger,induces autophagy by inhibiting the AKT/m TOR pathway in human non-small cell lung cancer A549 cells[J].J.Agric.Food Chem.,2009,57(20):9809-9816
[22]Tan B S,Kang O,Mai C W,et al.6-Shogaol inhibits breast and colon cancer cell proliferation through activation of peroxisomal proliferator activated receptor gamma(PPAR gamma)[J].Cancer Lett.,2013,336:127-139
[23]Park S J,Lee M Y,Son B S,et al.TBK1-targeted suppression of TRIF-dependent signaling pathway of Toll-like receptors by 6-shogaol,an active component of ginger[J].Biosci.,Biotechnol.,Biochem.,2009,73(7):1474-1478