WANG Xiao-bo,XIONG Xiao-wei,ZHU Yu-hua,et al.Effects of Daidzein and Its Phenylsulfonate Derivatives on Proliferation and Apoptosis of Lung Cancer Cells[J].Soybean Science,2019,38(06):934-942.[doi:10.11861/j.issn.1000-9841.2019.06.0934]
大豆苷元及其苯磺酸酯衍生物对肺癌细胞增殖和凋亡的影响
- Title:
- Effects of Daidzein and Its Phenylsulfonate Derivatives on Proliferation and Apoptosis of Lung Cancer Cells
- 摘要:
- 为研究大豆苷元及其3种苯磺酸酯衍生物对非小细胞肺癌细胞的增殖和凋亡的影响，利用CCK-8检测大豆苷元及衍生物处理A549、H1299和HELF细胞后细胞活性的时间-效应关系和剂量-效应关系，并通过Tunel流式细胞术检测衍生物B处理的细胞凋亡情况，RT-qPCR分析TP53和Caspase9 mRNA表达。结果表明：在1.0×10-5~1.0×103 μmol·L-1浓度范围时，加入大豆苷元的浓度每增加10倍，肺正常HELF细胞的活性增加16.11%［95%CI 13.74，18.49］(P<0.001)，肺癌A549与H1299细胞活性则分别降低3.26% (P<0.001)、3.33% (P<0.001)；加入衍生物B的浓度每增加10倍，HELF细胞的活性增加9.92% (P<0.001)，A549和H1299细胞活性降低5.23% (P<0.001)、4.32%（P<0.001）。10 μmol·L-1大豆苷元浓度下，作用时间每延长1 h，HELF细胞活性增加1.87%（P<0.001），A549和H1299细胞活性分别降低1.30%（P<0.001）、1.75%（P<0.001），苯磺酸酯衍生物B对HELF细胞活性增加1.72%（P<0.001），A549和H1299细胞降低1.46%（P<0.001）、2.07%（P=0.001）。10 μmol·L-1的衍生物B处理6 h后，HELF细胞相对数量增加了42.66%（P=0.03），A549和H1299细胞相对数量减少20.67%（P=0.035）、18.33%（P=0.043）。衍生物B促进A549和H1299细胞的凋亡的作用约为对照组的1.15倍（P<0.001）、1.24倍（P=0.001），抑制HELF细胞的凋亡作用为对照组的0.84倍（P=0.005），10 μmol·L-1衍生物B处理时肺癌和肺正常细胞的TP53和Caspase9的mRNA表达比对照组高。研究表明：大豆苷元及其3种苯磺酸酯衍生物能够抑制肺癌细胞的增殖活性的同时，增加肺正常细胞增殖活性，大豆苷元苯磺酸酯衍生物B能促进肺癌细胞凋亡并抑制肺正常细胞凋亡，其机制可能与通过P53通路诱导癌细胞凋亡有关。
- Abstract:
- In order to study the effects of daidzeint and their three phenysulfonate derivatives on proliferation and apoptosis of non-small cell lung cancer （NSCLC）cells, in vitro experiments were conducted on the treatment of A549, H1299 and HELF cells by daidzein and their three derivatives with 1.0×10-5-1.0×103 μmol·L-1 and 0-24 h treatment. CCK-8 was used to detect cell activity and apoptosis was detected by Tunel assay. RT-qPCR was used to verify the speculate that these effects maybe involved in TP53 and Caspase9. The results showed that, for the concentration range of 1×10-5-1×103 μmol·L-1, when the concentration of daidzein and derivative B was increased 10 times repectively, the proliferation of lung cancer cells could be inhibited to 3.26% (P<0.001), 5.23% (P<0.001) in A549 cells and 3.33% (P<0.001), 4.32%（P<0.001）in H1299 cells, while they were promoted 16.11% ［95% CI 13.74,18.49］(P<0.001), 9.92% (P<0.001) in HELF cells. By treatment with 10 μmol·L-1 of daidzein and derivative B from 0 to 24 h, HELF cells were promoted to 1.87%（P<0.001）, 1.72%（P<0.001）for every hour extent, while A549 cells were inhibited to 1.30%（P<0.001）, 1.46%（P<0.001） and H1299 cells were inhibited to 1.75%（P<0.001）, 2.07%（P=0.001）respectively. The relative counts of HELF cells increased by 42.66%（P=0.03）, and the counts of H1299 cells decreased by 18.33%（P=0.043）and A549 cells decreased by 20.67%（P=0.035）, when treated with 10 μmol·L-1 derivative B for 6 h. 10 μmol·L-1 derivative B could promote A549 and H1299 cells to apoptosis with 1.15 times（P<0.001） and 1.24 times (P=0.001), and protect HELF cells out of apoptosis with 0.84 times（P=0.005）. The mRNA expression of TP53and Caspase9were up-regulation in three cells. To a conclusion, daidzein and their 3 derivatives can inhibit the proliferation of lung cancer cells, and the effect is related to the concentration and time of treatment. derivative B can promote the apoptosis of lung cancer cells. The proliferation of lung cancer cells may be associated with the induction of apoptosis such as TP53and Caspase9.
参考文献/References:
［1］Szeja W, Grynkiewicz G, Rusin A. Isoflavones, their glycosides and glycoconjugates. Synthesis and Biological Activity［J］. Current Organic Chemistry, 2017, 21(3): 218-235. ［2］Zaheer K, Humayoun A M. An updated review of dietary isoflavones: Nutrition, processing, bioavailability and impacts on human health［J］. Critical Reviews in Food Science and Nutrition, 2017, 57(6): 1280-1293. ［3］郑峰, 金延华, 何鑫, 等. 大豆异黄酮和皂甙对H22小鼠肝癌移植瘤的生长抑制及促细胞凋亡作用［J］. 大豆科学, 2016, 35(3): 494-497.(Zheng F, Jin Y H, He X, et al. Anti-tumor and pro-apoptotic effects of soy isoflavones and saponins on h22-bearing mice［J］. Soybean Science, 2016, 35(3): 494-497.)［4］Peng Y, Shi Y, Zhang H, et al. Anti-inflammatory and anti-oxidative activities of daidzein and its sulfonic acid ester derivatives［J］. Journal of Functional Foods, 2017, 35: 635-640.［5］Reger M K, Zollinger T W, Liu Z, et al. Dietary intake of isoflavones and coumestrol and the risk of prostate cancer in the prostate, lung, colorectal and ovarian cancer screening trial［J］. International Journal of Cancer, 2018, 142(4): 719-728. ［6］彭游, 付小兰, 邓泽元. 新型大豆异黄酮磺酸酯的临床前药物动力学［J］. 时珍国医国药, 2011, 22(12): 2932-2935. (Peng Y, Fu X L, Deng Z Y. Pre-clinical pharmacokinetics of novel soybean iosflavone sulfonate［J］. Lishizhen Medicine and Materia Medica Research, 2011, 22(12): 2932-2935. )［7］彭游, 邓泽元, 叶兴琳. 异黄酮衍生物的设计, 合成与抗肿瘤活性［J］. 天然产物研究与开发, 2009, 21(5): 796-799. (Peng Y, Deng Z Y, Ye X L. Design, synthesis and anticancer activity of isofavone derivatives［J］. Natural Product Rsearch and Development, 2009, 21(5): 796-799. )［8］He Y, Wu X, Cao Y, et al. Daidzein exerts anti-tumor activity against bladder cancer cells via inhibition of FGFR3 pathway［J］. Neoplasma, 2016, 63(4): 523-531. ［9］Bray F, Ferlay J, Soerjomataram I, et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries［J］. CA: A Cancer Journal for Clinicians, 2018, 68(6): 394-424. ［10］Ferlay J, Colombet M, Soerjomataram I, et al. Estimating the global cancer incidence and mortality in 2018: GLOBOCAN sources and methods［J］. International Journal of Cancer, 2019.114(8):1941-1953. ［11］Applegate C, Rowles J, Ranard K, et al. Soy consumption and the risk of prostate cancer: An updated systematic review and meta-analysis［J］. Nutrients, 2018, 10(1): 40-65. ［12］Zhao T, Jin F, Li J, et al. Dietary isoflavones or isoflavone-rich food intake and breast cancer risk: A meta-analysis of prospective cohort studies［J］. Clinical Nutrition, 2019, 38(1): 136-145. ［13］Jin H, Leng Q, Li C. Dietary flavonoid for preventing colorectal neoplasms［J］. Cochrane Database of Systematic Reviews, 2012, 8(8): D009350. ［14］Pistritto G, Trisciuoglio D, Ceci C, et al. Apoptosis as anticancer mechanism: Function and dysfunction of its modulators and targeted therapeutic strategies［J］. Aging, 2016, 8(4): 603-619. ［15］Rasmussen M A, Holst B R, Tümer Z, et al. Transient p53 suppression increases reprogramming of human fibroblasts without affecting apoptosis and DNA damage［J］. Stem Cell Reports, 2014, 3(3): 404-413. ［16］Kim B, Srivastava S K, Kim S H. Caspase-9 as a therapeutic target for treating cancer［J］. Expert Opinion on Therapeutic Targets, 2015, 19(1): 113-127. ［17］Jiang X, Cao C, Sun W, et al. Scandenolone from Cudrania tricuspidata fruit extract suppresses the viability of breast cancer cells (MCF-7) in vitro and in vivo［J］. Food and Chemical Toxicology, 2019, 126(4): 56-66. ［18］曾靖, 魏桂林, 钟星明, 等. 3′-大豆苷元磺酸钠体外抗肿瘤活性的筛选［J］. 赣南医学院学报, 2017, 37(3): 351-353. (Zeng J, Wei G L, Zhong X M, et al. In vitro anti-tumor activity screening of 3′-daidzein sulfonate sodium［J］. Journal of Gannan Medical University, 2017, 37(3): 351-353. )［19］Ahn S, Jo M S, Lee D, et al. Dual effects of isoflavonoids from Pueraria lobata roots on estrogenic activity and anti-proliferation of MCF-7 human breast carcinoma cells［J］. Bioorganic Chemistry, 2019, 83(3): 135-144. ［20］Ko K, Yeo Y, Yoon J, et al. Plasma phytoestrogens concentration and risk of colorectal cancer in two different Asian populations［J］. Clinical Nutrition, 2018, 37(5): 1675-1682. ［21］Shokoohinia Y, Hosseinzadeh L, Alipour M, et al. Comparative evaluation of cytotoxic and apoptogenic effects of several coumarins on human cancer cell lines: Osthole induces apoptosis in p53-deficient H1299 cells［J］. Advances in Pharmacological Sciences, 2014: 1-8. ［22］Yun H S, Baek J, Yim J, et al. Knockdown of hepatoma-derived growth factor-related protein-3 induces apoptosis of H1299 cells via ROS-dependent and p53-independent NF-κB activation［J］. Biochemical and Biophysical Research Communications, 2014, 449(4): 471-476. ［23］Gadepalli V S, Deb S P, Deb S, et al. Lung cancer stem cells, p53 mutations and MDM2［J］. Subcellular Biochemistry, 2014, 85: 359-370. ［24］金时. 大豆苷元体外诱导乳腺癌细胞凋亡的线粒体-caspase途径研究［D］. 哈尔滨：哈尔滨医科大学, 2010. (Jin S. Daidzein induces breast cancer cell apoptosis via the mitochondrial-caspasse dependent pathway［D］. Harbin：Harbin Medical University, 2010. )
相似文献/References:
[1]崔力剑,石素琴,刘敏彦,等.淡豆豉中多指标成分的含量测定方法研究[J].大豆科学,2012,31(01):127.[doi:10.3969/j.issn.1000-9841.2012.01.029]
CUI li-jian,SHI Su-qin,LIU Min-yan,et al.Determination Method of Multi-target Components in Semen Sojae Praeparatum[J].Soybean Science,2012,31(06):127.[doi:10.3969/j.issn.1000-9841.2012.01.029]
[2]曲嘉,刘章雄,李开盛,等.多年生野生大豆的生物学性状及叶片中大豆苷元和染料木素含量分析[J].大豆科学,2008,27(06):949.[doi:10.11861/j.issn.1000-9841.2008.06.0949]
QU Jia,LIU Zhang-xiong,LI Kai-sheng,et al.Anaylsis of Biological Traits and Daidzein and Genistein Content of Perennial Wild Soybean(GlycineL.Subgenus Glycine)[J].Soybean Science,2008,27(06):949.[doi:10.11861/j.issn.1000-9841.2008.06.0949]
[3]牛丽颖,杜红娜,刘姣,等.淡豆豉炮制前后异黄酮组分含量的比较[J].大豆科学,2008,27(04):672.[doi:10.11861/j.issn.1000-9841.2008.04.0672]
NIU Li-ying,DU Hong-na,LIU Jiao,et al.Comparative Analysis of Isoflavone Contents in Semen Sojae Praeparatum Before and After Processing[J].Soybean Science,2008,27(06):672.[doi:10.11861/j.issn.1000-9841.2008.04.0672]
备注/Memo
基金项目：国家自然科学基金（81360447，81660535，81660541）。第一作者简介：王晓波 （1990-），男，硕士，主要从事流行病与卫生统计学研究。E-mail：1181894471@qq.com。通讯作者：汪鑫（1966-），男，硕士，教授，主要从事分子流行病学研究。E-mail：wangxin0072@126.com；黄邵鑫（1985-），男，博士，副教授，主要从事化合物健康评价研究。E-mail：517159865@qq.com。