CLC number: Q813
On-line Access: 2024-08-27
Received: 2023-10-17
Revision Accepted: 2024-05-08
Crosschecked: 2017-11-22
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Mei-lin Cui, Huan-yi Yang, Guo-qing He. Apoptosis induction of colorectal cancer cells HTL-9 in vitro by the transformed products of soybean isoflavones by Ganoderma lucidum[J]. Journal of Zhejiang University Science B,in press.Frontiers of Information Technology & Electronic Engineering,in press.https://doi.org/10.1631/jzus.B1700189 @article{title="Apoptosis induction of colorectal cancer cells HTL-9 in vitro by the transformed products of soybean isoflavones by Ganoderma lucidum", %0 Journal Article TY - JOUR
灵芝菌生物转化大豆异黄酮及其产物对结直肠癌细胞HTL-9的体外凋亡诱导研究创新点:灵芝是一种珍贵的药用真菌,大豆异黄酮的苷元物质也具有重要的药理活性,本文首次利用灵芝菌液体发酵的匀浆液生物转化大豆异黄酮,所得到的产物中大豆苷元与染料木素转化率高,同时还富集了灵芝菌的活性成分,并对转化产物的抗癌活性及机理进行了初步探讨。 方法:首先利用灵芝菌液体发酵的匀浆体系生物转化大豆异黄酮(图1)。其次,对转化产物的抗癌活性进行研究,主要包括对癌细胞存活率(图2)、细胞凋亡(图3)及细胞周期分布(图4)的影响。最后,利用蛋白质印迹(Western-blot)与逆转录聚合酶链反应(RT-PCR)技术对凋亡相关的基因和蛋白进行检测(图5和表2),初步探讨转化产物的体外抗癌机理。 结论:本实验结果显示,转化产物中大豆苷元及染料木素的转化率分别为96.63%和87.82%,其中染料木素的含量可达(703.21±4.35) mg/g,同时转化产物中还富含了灵芝菌的活性成分。其次,对转化产物抗癌活性研究发现,其能有效降低HTL-9细胞的存活率,可通过将细胞阻滞于G1期而诱导细胞晚期凋亡。此外,转化产物(100 µg/ml)还可明显上调Bax、Caspase-3、Caspase-8、Cyto-c和p53的表达量,而Survivin和NF-κB表达量发生明显下调。结果表明,转化产物主要通过线粒体途径诱导细胞凋亡,但同时还调控多个与凋亡相关的基因。 关键词组: Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article
Reference[1]Al-Fatlawi, A.A., Abbas, A., Zafaryab, M., et al., 2014. Rhein induced cell death and apoptosis through caspase dependent and associated with modulation of p53, Bcl-2/Bax ratio in human cell lines. Int. J. Pharm. Pharmaceut. Sci., 6(2):515-519. ![]() [2]Andlauer, W., Kolb, J., Stehle, P., et al., 2000. Absorption and metabolism of genistein in isolated rat small intestine. J. Nutr., 130(4):843-846. ![]() [3]Baglia, M.L., Gu, K., Zhang, X., et al., 2015. Soy isoflavone intake and bone mineral density in breast cancer survivors. Cancer Causes Control, 26(4):571-580. ![]() [4]Banerjee, S., Ali, S., Azmi, A., et al., 2012. Abstract 2698: improved therapeutic activity of isoflavone-G2535 and docetaxel combination in hormone refractory prostate cancer. Cancer Res., 72(8 Suppl.):2698. ![]() [5]Budhathoki, S., Joshi, A.M., Ohnaka, K., et al., 2011. Soy food and isoflavone intake and colorectal cancer risk: the fukuoka colorectal cancer study. Scand. J. Gastroenterol., 46(2):165-172. ![]() [6]Chen, J., Hou, R., Zhang, X., et al., 2014. Calycosin suppresses breast cancer cell growth via ERβ-dependent regulation of IGF-1R, p38 MAPK and PI3K/Akt pathways. PLoS ONE, 9(3):e91245. ![]() [7]Choi, E.J., Kim, G.H., 2013. Antiproliferative activity of daidzein and genistein may be related to ERα/c-erbB-2 expression in human breast cancer cells. Mol. Med. Rep., 7(3):781-784. ![]() [8]Cotrim, C.Z., Fabris, V., Doria, M.L., et al., 2013. Estrogen receptor β growth-inhibitory effects are repressed through activation of MAPK and PI3K signalling in mammary epithelial and breast cancer cells. Oncogene, 32(19):2390-2402. ![]() [9]Cui, M.L., Yang, H.Y., He, G.Q., 2015. Submerged fermentation production and characterization of intracellular triterpenoids from Ganoderma lucidum using HPLC-ESI-MS. J. Zhejiang Univ.-Sci. B (Biomed. & Biotechnol.), 16(12):998-1010. ![]() [10]Ewe, J.A., Wan-Abdullah, W.N., Alias, A.K., et al., 2012. Enhanced growth of lactobacilli and bioconversion of isoflavones in biotin-supplemented soymilk by electroporation. Int. J. Food Sci. Nutr., 63(5):580-596. ![]() [11]Ferlay, J., Soerjomataram, I., Dikshit, R., et al., 2015. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int. J. Cancer, 136(5):E359-E386. ![]() [12]Gong, Y., Li, Y., Lu, Y., et al., 2011. Bioactive tanshinones in Salvia miltiorrhiza inhibit the growth of prostate cancer cells in vitro and in mice. Int. J. Cancer, 129(5):1042-1052. ![]() [13]Guo, X.Y., Liu, D., Ye, M., et al., 2013. Structural characterization of minor metabolites and pharmacokinetics of ganoderic acid C2 in rat plasma by HPLC coupled with electrospray ionization tandem mass spectrometry. J. Pharm. Biomed. Anal., 75:64-73. ![]() [14]Handa, C.L., Couto, U.R., Vicensoti, A.H., et al., 2014. Optimisation of soy flour fermentation parameters to produce beta-glucosidase for bioconversion into aglycones. Food Chem., 152:56-65. ![]() [15]Hati, S., Vij, S., Singh, B.P., et al., 2015. β-Glucosidase activity and bioconversion of isoflavones during fermentation of soymilk. J. Sci. Food Agric., 95(1):216-220. ![]() [16]Hsin, I.L., Ou, C.C., Wu, M.F., et al., 2015. GMI, an immunomodulatory protein from Ganoderma microsporum, potentiates cisplatin-induced apoptosis via autophagy in lung cancer cells. Mol. Pharm., 12(5):1534-1543. ![]() [17]Indran, I.R., Tufo, G., Pervaiz, S., et al., 2011. Recent advances in apoptosis, mitochondria and drug resistance in cancer cells. BBA-Bioenergetics, 1807(6):735-745. ![]() [18]Kang, D., Mutakin, M., Levita, J., 2015. Computational study of triterpenoids of Ganoderma lucidum with aspartic protease enzymes for discovering HIV-1 and plasmepsin inhibitors. Int. J. Chem., 7(1):62-68. ![]() [19]Keypour, S., Rafati, H., Riahi, H., et al., 2010. Qualitative analysis of ganoderic acids in Ganoderma lucidum from Iran and China by RP-HPLC and electrospray ionisation-mass spectrometry (ESI-MS). Food Chem., 119(4):1704-1708. ![]() [20]Kim, H.M., Paik, S.Y., Ra, K.S., et al., 2006. Enhanced production of exopolysaccharides by fed-batch culture of Ganoderma resinaceum DG-6556. J. Microbiol., 44(2):233-242. ![]() [21]Kurahashi, N., Iwasaki, M., Sasazuki, S., et al., 2007. Soy product and isoflavone consumption in relation to prostate cancer in Japanese men. Cancer Epidemiol. Biomarkers Prev., 16(3):538-545. ![]() [22]Lee, S.Y., Rhee, H.M., 1990. Cardiovascular effects of mycelium extract of Ganoderma lucidum: inhibition of sympathetic outflow as a mechanism of its hypotensive action. Chem. Pharm. Bull., 38(5):1359-1364. ![]() [23]Li, Y., Kong, D., Ahmad, A., et al., 2012a. Epigenetic deregulation of miR-29a and miR-1256 by isoflavone contributes to the inhibition of prostate cancer cell growth and invasion. Epigenetics, 7(8):940-949. ![]() [24]Li, Y., Kong, D., Ahmad, A., et al., 2012b. Targeting bone remodeling by isoflavone and 3,3’-diindolylmethane in the context of prostate cancer bone metastasis. PLoS ONE, 7(3):e33011. ![]() [25]Lim, J.C.W., Chan, T.K., Ng, D.S., et al., 2012. Andrographolide and its analogues: versatile bioactive molecules for combating inflammation and cancer. Clin. Exp. Pharmacol. Physiol., 39(3):300-310. ![]() [26]Liu, F., Bardhan, K., Yang, D., et al., 2012. NF-κB directly regulates Fas transcription to modulate Fas-mediated apoptosis and tumor suppression. J. Biol. Chem., 287(30):25530-25540. ![]() [27]Liu, J., Zhang, C., Hu, W., et al., 2015. Tumor suppressor p53 and its mutants in cancer metabolism. Cancer Lett., 356(2):197-203. ![]() [28]Loganathan, J., Jiang, J., Smith, A., et al., 2014. The mushroom Ganoderma lucidum suppresses breast-to-lung cancer metastasis through the inhibition of pro-invasive genes. Int. J. Oncol., 44(6):2009-2015. ![]() [29]Maitan-Alfenas, G.P., Lorena, G.A., de Almeida, M.N., et al., 2014. Hydrolysis of soybean isoflavones by Debaryomyces hansenii UFV-1 immobilised cells and free β-glucosidase. Food Chem., 146:429-436. ![]() [30]Mayola, E., Gallerne, C., Esposti, D.D., et al., 2011. Withaferin a induces apoptosis in human melanoma cells through generation of reactive oxygen species and down-regulation of Bcl-2. Apoptosis, 16(10):1014-1027. ![]() [31]Mense, S.M., Hei, T.K., Ganju, R.K., et al., 2008. Phytoestrogens and breast cancer prevention: possible mechanisms of action. Environ. Health Perspect., 116(4):426-433. ![]() [32]Miranda, C.F., Morales-Cruz, M., Suarez, B., et al., 2014. Effect of cytochrome c modification with co-polymer on its apoptotic activity for cancer treatment (LB248). FASEB J., 28(1 Suppl.):248. ![]() [33]Munck, L., Jorgensen, K.G., Ruud-Hansen, J., et al., 1989. The EBC methods for determination of high molecular weight β-glucan in barley, malt, wort and beer. J. Inst. Brewing, 95(2):79-82. ![]() [34]Ollberding, N.J., Lim, U., Wilkens, L.R., et al., 2012. Legume, soy, tofu, and isoflavone intake and endometrial cancer risk in postmenopausal women in the multiethnic cohort study. J. Natl. Cancer Inst., 104(1):67-76. ![]() [35]Prietsch, R.F., Monte, L.G., da Silva, F.A., et al., 2014. Genistein induces apoptosis and autophagy in human breast MCF-7 cells by modulating the expression of proapoptotic factors and oxidative stress enzymes. Mol. Cell Biochem., 390(1-2):235-242. ![]() [36]Priyadarsini, R.V., Murugan, R.S., Maitreyi, S., et al., 2010. The flavonoid quercetin induces cell cycle arrest and mitochondria-mediated apoptosis in human cervical cancer (HeLa) cells through p53 induction and NF-κB inhibition. Eur. J. Pharmacol., 649(1-3):84-91. ![]() [37]Selent, J., Kaczor, A.A., Guixa-Gonzalez, R., et al., 2013. Rational design of the survivin/CDK4 complex by combining protein-protein docking and molecular dynamics simulations. J. Mol. Model., 19(4):1507-1514. ![]() [38]Srivastava, K., Singh, A.K., Khan, K., et al., 2014. Assessment of enhancement of peak bone gain by isoflavone enriched standardized soy extract in female rats. J. Funct. Foods, 7:314-321. ![]() [39]Suarez-Jimenez, G.M., Burgos-Hernandez, A., Ezquerra-Brauer, J.M., 2012. Bioactive peptides and depsipeptides with anticancer potential: sources from marine animals. Marine Drugs, 10(12):963-986. ![]() [40]Szliszka, E., Czuba, Z.P., Sędek, L., et al., 2011. Enhanced TRAIL-mediated apoptosis in prostate cancer cells by the bioactive compounds neobavaisoflavone and psoralidin isolated from Psoralea corylifolia. Pharm. Rep., 63(1):139-148. ![]() [41]Tang, W., Liu, J.W., Zhao, W.M., et al., 2006. Ganoderic acid T from Ganoderma lucidum mycelia induces mitochondria mediated apoptosis in lung cancer cells. Life Sci., 80(3):205-211. ![]() [42]Thomas, S., Quinn, B.A., Das, S.K., et al., 2013. Targeting the Bcl-2 family for cancer therapy. Exp. Opin. Therapeut. Tar., 17(1):61-75. ![]() [43]Titiek, F., Umar, S., Cahyanto, M., et al., 2013. Effect of indigenous lactic acid bacteria fermentation on enrichment of isoflavone and antioxidant properties of kerandang (Canavalia virosa) extract. Int. Food Res. J., 20(5):2945-2950. ![]() [44]Tse, G., Eslick, G.D., 2016. Soy and isoflavone consumption and risk of gastrointestinal cancer: a systematic review and meta-analysis. Eur. J. Nutr., 55(1):63-73. ![]() [45]Tsuboy, M.S., Marcarini, J.C., de Souza, A.O., et al., 2014. Genistein at maximal physiologic serum levels induces G0/G1 arrest in MCF-7 and HB4a cells, but not apoptosis. J. Med. Food, 17(2):218-225. ![]() [46]van Meerloo, J., Kaspers, G.J., Cloos, J., 2011. Cell sensitivity assays: the MTT assay. In: Cree, I. (Ed.), Cancer Cell Culture. Methods in Molecular Biology (Methods and Protocols), Vol. 731. Humana Press, p.237-245. ![]() [47]Varfolomeev, E., Goncharov, T., Vucic, D., 2015. Roles of c-IAP proteins in TNF receptor family activation of NF-κB signaling. In: May, M. (Ed.), NF-κ B. Methods in Molecular Biology, Vol. 1280. Humana Press, New York, p.269-282. ![]() [48]Wei, J., Bhatt, S., Chang, L.M., et al., 2012. Isoflavones, genistein and daidzein, regulate mucosal immune response by suppressing dendritic cell function. PLoS ONE, 7(10):e47979. ![]() [49]Xu, C.F., Wu, A.R., Zhu, H., et al., 2013. Melatonin is involved in the apoptosis and necrosis of pancreatic cancer cell line SW-1990 via modulating of Bcl-2/Bax balance. Biomed. Pharmacother., 67(2):133-139. ![]() [50]Yan, L., Spitznagel, E.L., Bosland, M.C., 2010. Soy consumption and colorectal cancer risk in humans: a meta-analysis. Cancer Epidem. Biomar. Prev., 19(1):148-158. ![]() [51]Yeo, S.K., Liong, M.T., 2010. Angiotensin I-converting enzyme inhibitory activity and bioconversion of isoflavones by probiotics in soymilk supplemented with prebiotics. Int. J. Food Sci. Nutr., 61(2):161-181. ![]() [52]Yeom, S.J., Kim, B.N., Kim, Y.S., et al., 2012. Hydrolysis of isoflavone glycosides by a thermostable beta-glucosidase from Pyrococcus furiosus. J. Agric. Food Chem., 260(6):1535-1541. ![]() [53]Yin, L.J., Tai, H.M., Lee, H.H., et al., 2014. Proteolysis and lactobacillus fermentation effects on the isoflavones transformation and removal of anti-nutritional factors of soy bean. J. Mar. Sci. Technol., 22(4):525-530. ![]() [54]Zhao, W., Xu, J.W., Zhong, J.J., 2011. Enhanced production of ganoderic acids in static liquid culture of Ganoderma lucidum under nitrogen-limiting conditions. Bioresour. Technol., 102(17):8185-8190. ![]() [55]Zhou, C., Lin, H., Ge, X., et al., 2015. 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