5-allyl-7-gen-difluoromethoxychrysin enhances TRAIL-induced apoptosis in human lung carcinoma A549 cells

BMC Cancer

Zhao-Hui Xie¹  Mei-Fang Quan¹  Fei Liu¹  Jian-Guo Cao¹  Jian-Song Zhang¹ 

[1] Medical College, Hunan Normal University, Changsha 410013, Hunan Province, China

关键词: therapeutic action;    apoptosis;    TRAIL;    5-allyl-7-gen-difluoromethoxychrysin;    chrysin;    Lung cancer;   

Others  :  1080813
DOI  :  10.1186/1471-2407-11-322

 received in 2010-12-24, accepted in 2011-07-29,  发布年份 2011

【 摘 要 】

Background

5-allyl-7-gen-difluoromethoxychrysin (AFMC) is a novel synthetic analogue of chrysin that has been reported to inhibit proliferation in various cancer cell lines. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anti-cancer agent.

Methods

The cytotoxicity of A549 and WI-38 cells were determined using colorimetry. Apoptosis was detected by flow cytometry (FCM) after propidium iodide (PI) fluorescence staining and agarose gel electrophoresis. Caspase activities were evaluated using enzyme-linked immunosorbent assay (ELISA).The expressions of DR4 and DR5 were analyzed using FCM and western blot.

Results

Subtoxic concentrations of AFMC sensitize human non-small cell lung cancer (NSCLC) A549 cells to TRAIL-mediated apoptosis. Combined treatment of A549 cells with AFMC and TRAIL significantly activated caspase-3, -8 and -9. The caspase-3 inhibitor zDEVD-fmk and the caspase-8 inhibitor zIETD-fmk blocked the apoptosis of A549 cells induced by co-treatment with AFMC and TRAIL. In addition, we found that treatment of A549 cells with AFMC significantly induced the expression of death receptor 5 (DR5). AFMC-mediated sensitization of A549 cells to TRAIL was efficiently reduced by administration of a blocking antibody or small interfering RNAs against DR5. AFMC also caused increase of the Sub-G1 cells by TRAIL treatment and increased the expression levels of DR5 in other NSCLC H460 and H157 cell lines. In contrast, AFMC-mediated induction of DR5 expression was not observed in human embryo lung WI-38 cells, and AFMC did not sensitize WI-38 cells to TRAIL-induced apoptosis.

Conclusions

AFMC synergistically enhances TRAIL-mediated apoptosis in NSCLC cells through up-regulating DR5 expression.

【 授权许可】

   
2011 Xie et al; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Densmore CL: Advances in noninvasive pulmonary gene therapy. Curr Drug Deliv 2006, 3:55-63.
• [2]Wang C, Li Z, Lu Y, Du R, Katiyar S, Yang J, Fu M, Leader JE, Quong A, Novikoff PM, Pestell RG: Cyclin D1 repression of nuclear respiratory factor 1 integrates nuclear DNA synthesis and mitochondrial function. Proc Natl Acad Sci USA 2006, 103:11567-11572.
• [3]Hong TB, Rahumatullah A, Yogarajah T, Ahmad M, Yin KB: Potential effects of chrysin on MDA-MB-231 cells. Int J Mol Sci 2010, 11:1057-1069.
• [4]Khoo BY, Chua SL, Balaram P: Apoptotic effects of chrysin in human cancer cell lines. Int J Mol Sci 2010, 11:2188-2199.
• [5]Woo KJ, Jeong YJ, Park JW, Kwon TK: Chrysin-induced apoptosis is mediated through caspase activation and Akt inactivation in U937 leukemia cells. Biochem Biophys Res Commun 2004, 325:1215-1222.
• [6]Zheng X, Cao JG, Liao DF, Liu HT: Systhesis and Anticancer Effect of gem-Difluoromethylenated Chrysin Derivatives. Chinese Chemical Letter 2006, 17:1439-1442.
• [7]Zheng X, Meng WD, Xu YY, Cao JG, Qing FL: Synthesis and anticancer effect of chrysin derivatives. Bioorg Med Chem Lett 2003, 13:881-884.
• [8]Ai XH, Zheng X, Tang XQ, Sun L, Zhang YQ, Qin Y, Liu HQ, Xia H, Cao JG: Induction of apoptosis of human gastric carcinoma SGC-7901 cell line by 5, 7-dihydroxy-8-nitrochrysin in vitro. World J Gastroenterol 2007, 13:3824-3828.
• [9]Yang XH, Zheng X, Cao JG, Xiang HL, Liu F, Lv Y: 8-Bromo-7-methoxychrysin-induced apoptosis of hepatocellular carcinoma cells involves ROS and JNK. World J Gastroenterol 2010, 16:3385-3393.
• [10]Li JL, Xie WY, Cao JG: The Effect of 5-allyl-7-gendifluoromethylenechrysin on proliferation and apoptosis in ovarian cancer Cell Cultured in Vitro. Am J Chin Clin Med 2005, 7:323-326.
• [11]Tan XW, Xia H, Xu JH, Cao JG: Induction of apoptosis in human liver carcinoma HepG2 cell line by 5-allyl-7-gen-difluoromethylenechrysin. World J Gastroenterol 2009, 15:2234-2239.
• [12]Mariani SM, Krammer PH: Differential regulation of TRAIL and CD95 ligand in transformed cells of the T and B lymphocyte lineage. Eur J Immunol 1998, 28:973-982.
• [13]Bodmer JL, Meier P, Tschopp J, Schneider P: Cysteine 230 is essential for the structure and activity of the cytotoxic ligand TRAIL. J Biol Chem 2000, 275:20632-20637.
• [14]Spierings DC, de Vries EG, Vellenga E, van den Heuvel FA, Koornstra JJ, Wesseling J, Hollema H, de Jong S: Tissue distribution of the death ligand TRAIL and its receptors. J Histochem Cytochem 2004, 52:821-831.
• [15]Sheridan JP, Marsters SA, Pitti RM, Gurney A, Skubatch M, Baldwin D, Ramakrishnan L, Gray CL, Baker K, Wood WI, Goddard AD, Godowski P, Ashkenazi A: Control of TRAIL-induced apoptosis by a family of signaling and decoy receptors. Science 1997, 277:818-821.
• [16]MacFarlane M, Ahmad M, Srinivasula SM, Fernandes-Alnemri T, Cohen GM, Alnemri ES: Identification and molecular cloning of two novel receptors for the cytotoxic ligand TRAIL. J Biol Chem 1997, 272:25417-25420.
• [17]Ashkenazi A, Dixit VM: Death receptors: signaling and modulation. Science 1998, 281:1305-1308.
• [18]Takimoto R, El-Deiry WS: Wild-type p53 transactivates the KILLER/DR5 gene through an intronic sequence-specific DNA-binding site. Oncogene 2000, 19:1735-1743.
• [19]Almasan A, Ashkenazi A: Apo2L/TRAIL: apoptosis signaling, biology, and potential for cancer therapy. Cytokine Growth Factor Rev 2003, 14:337-348.
• [20]Walczak H, Haas TL: Biochemical analysis of the native TRAIL death-inducing signaling complex. Methods Mol Biol 2008, 414:221-239.
• [21]Corazza N, Kassahn D, Jakob S, Badmann A, Brunner T: TRAIL-induced apoptosis: between tumor therapy and immunopathology. Ann N Y Acad Sci 2009, 1171:50-58.
• [22]Zhang L, Fang B: Mechanisms of resistance to TRAIL-induced apoptosis in cancer. Cancer Gene Ther 2005, 12:228-237.
• [23]Thorburn A, Behbakht K, Ford H: TRAIL receptor-targeted therapeutics: resistance mechanisms and strategies to avoid them. Drug Resist Updat 2008, 11:17-24.
• [24]Szliszka E BJ MA, Miszkiewicz J, Krol W: Enhanced sensitivity of hormone-refractory prostate cancer cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) mediated cytotoxicity by taxanes. CEJ Urol 2009, 62:29-34.
• [25]Szliszka E, Czuba ZP, Bronikowska J, Mertas A, Paradysz A, Krol W: Ethanolic Extract of Propolis Augments TRAIL-Induced Apoptotic Death in Prostate Cancer Cells. Evid Based Complement Alternat Med 2009.
• [26]Szliszka E CZ MB, Sedek L, Paradysz A, Krol W: Chalcones enhances TRAIL-induced apoptosis in prostate cancer cells. Int J Mol Sci 2010, 11:1-13.
• [27]Nakata S, Yoshida T, Horinaka M, Shiraishi T, Wakada M, Sakai T: Histone deacetylase inhibitors upregulate death receptor 5/TRAIL-R2 and sensitize apoptosis induced by TRAIL/APO2-L in human malignant tumor cells. Oncogene 2004, 23:6261-6271.
• [28]Horinaka M, Yoshida T, Shiraishi T, Nakata S, Wakada M, Sakai T: The dietary flavonoid apigenin sensitizes malignant tumor cells to tumor necrosis factor-related apoptosis-inducing ligand. Mol Cancer Ther 2006, 5:945-951.
• [29]Qiu Y, Liu X, Zou W, Yue P, Lonial S, Khuri FR, Sun SY: The farnesyltransferase inhibitor R115777 up-regulates the expression of death receptor 5 and enhances TRAIL-induced apoptosis in human lung cancer cells. Cancer Res 2007, 67:4973-4980.
• [30]Chen W, Wang X, Zhuang J, Zhang L, Lin Y: Induction of death receptor 5 and suppression of survivin contribute to sensitization of TRAIL-induced cytotoxicity by quercetin in non-small cell lung cancer cells. Carcinogenesis 2007, 28:2114-2121.
• [31]Mitsudomi T, Steinberg SM, Nau MM, Carbone D, D'Amico D, Bodner S, Oie HK, Linnoila RI, Mulshine JL, Minna JD, Gazdar AF: p53 gene mutations in non-small-cell lung cancer cell lines and their correlation with the presence of ras mutations and clinical features. Oncogene 1992, 7:171-180.
• [32]Zhang YQ, Tang XQ, Sun L, Dong L, Qin Y, Liu HQ, Xia H, Cao JG: Rosiglitazone enhances fluorouracil-induced apoptosis of HT-29 cells by activating peroxisome proliferator-activated receptor gamma. World J Gastroenterol 2007, 13:1534-1540.
• [33]Ahmad KA, Harris NH, Johnson AD, Lindvall HC, Wang G, Ahmed K: Protein kinase CK2 modulates apoptosis induced by resveratrol and epigallocatechin-3-gallate in prostate cancer cells. Mol Cancer Ther 2007, 6:1006-1012.
• [34]Wang S, El-Deiry WS: Requirement of p53 targets in chemosensitization of colonic carcinoma to death ligand therapy. Proc Natl Acad Sci USA 2003, 100:15095-15100.
• [35]Liu X, Yue P, Zhou Z, Khuri FR, Sun SY: Death receptor regulation and celecoxib-induced apoptosis in human lung cancer cells. J Natl Cancer Inst 2004, 96:1769-1780.
• [36]Monasterio A, Urdaci MC, Pinchuk IV, Lopez-Moratalla N, Martinez-Irujo JJ: Flavonoids induce apoptosis in human leukemia U937 cells through caspase- and caspase-calpain-dependent pathways. Nutr Cancer 2004, 50:90-100.
• [37]Bodmer JL, Holler N, Reynard S, Vinciguerra P, Schneider P, Juo P, Blenis J, Tschopp J: TRAIL receptor-2 signals apoptosis through FADD and caspase-8. Nat Cell Biol 2000, 2:241-243.
• [38]Sprick MR, Weigand MA, Rieser E, Rauch CT, Juo P, Blenis J, Krammer PH, Walczak H: FADD/MORT1 and caspase-8 are recruited to TRAIL receptors 1 and 2 and are essential for apoptosis mediated by TRAIL receptor 2. Immunity 2000, 12:599-609.
• [39]Kischkel FC, Lawrence DA, Chuntharapai A, Schow P, Kim KJ, Ashkenazi A: Apo2L/TRAIL-dependent recruitment of endogenous FADD and caspase-8 to death receptors 4 and 5. Immunity 2000, 12:611-620.
• [40]Walle T, Ta N, Kawamori T, Wen X, Tsuji PA, Walle UK: Cancer chemopreventive properties of orally bioavailable flavonoids--methylated versus unmethylated flavones. Biochem Pharmacol 2007, 73:1288-1296.
• [41]Wen X, Walle T: Methylated flavonoids have greatly improved intestinal absorption and metabolic stability. Drug Metab Dispos 2006, 34:1786-1792.
• [42]Woo JH, Kim YH, Choi YJ, Kim DG, Lee KS, Bae JH, Min DS, Chang JS, Jeong YJ, Lee YH, Park JW, Kwon TK: Molecular mechanisms of curcumin-induced cytotoxicity: induction of apoptosis through generation of reactive oxygen species, down-regulation of Bcl-XL and IAP, the release of cytochrome c and inhibition of Akt. Carcinogenesis 2003, 24:1199-1208.
• [43]He Q, Huang Y, Sheikh MS: Proteasome inhibitor MG132 upregulates death receptor 5 and cooperates with Apo2L/TRAIL to induce apoptosis in Bax-proficient and -deficient cells. Oncogene 2004, 23:2554-2558.
• [44]Stinchcombe TE, Socinski MA: Considerations for second-line therapy of non-small cell lung cancer. Oncologist 2008, 13(Suppl 1):28-36.
• [45]Levine AJ: p53, the cellular gatekeeper for growth and division. Cell 1997, 88:323-331.
• [46]Greenblatt MS, Bennett WP, Hollstein M, Harris CC: Mutations in the p53 tumor suppressor gene: clues to cancer etiology and molecular pathogenesis. Cancer Res 1994, 54:4855-4878.
• [47]Srivastava RK, Kurzrock R, Shankar S: MS-275 sensitizes TRAIL-resistant breast cancer cells, inhibits angiogenesis and metastasis, and reverses epithelial-mesenchymal transition in vivo. Mol Cancer Ther 2010, 9:3254-3266.
• [48]Ganapathy S, Chen Q, Singh KP, Shankar S, Srivastava RK: Resveratrol enhances antitumor activity of TRAIL in prostate cancer xenografts through activation of FOXO transcription factor. PLoS One 2010, 5:e15627.
• [49]Chen Q, Ganapathy S, Singh KP, Shankar S, Srivastava RK: Resveratrol induces growth arrest and apoptosis through activation of FOXO transcription factors in prostate cancer cells. PLoS One 2010, 5:e15288.