藤黄酸衍生物-5诱导人头颈部鳞状细胞癌细胞凋亡的体外研究

›› 2016, Vol. 7 ›› Issue (1): 6-11.

藤黄酸衍生物-5诱导人头颈部鳞状细胞癌细胞凋亡的体外研究

李腾宇¹,²,潘劲松²,王旭³

1. 山东省泰安市中心医院口腔科
2. 上海交通大学医学院附属第一人民医院口腔科
3. 上海交通大学医学院附属第九人民医院口腔肿瘤实验室

收稿日期:2016-01-11 修回日期:2016-03-21 出版日期:2016-03-25 发布日期:2016-04-06
通讯作者: 王旭 E-mail:wxwxw2004@126.com

Gambogic acid derivative compound 5 induces apoptosis in human head and neck squamous cell carcinoma in vitro

Received:2016-01-11 Revised:2016-03-21 Online:2016-03-25 Published:2016-04-06

摘要/Abstract

摘要： [[摘要] 目的：研究藤黄酸（Gambogic acid, GA）的衍生物5（藤黄酸化合物5，C5），体外诱导人头颈部鳞状细胞癌（HNSCC）细胞的凋亡作用。方法：不同时间段分别用不同剂量C5作用于HNSCC细胞系CAL27、HN13和HN30，采用MTT法检测细胞存活率，Logit法测定抑制50%的细胞生长所需的浓度（IC50）值。采用Annexin-V/PI双染色流式细胞仪检测细胞凋亡。免疫细胞化学法和Western blot法检测凋亡相关蛋白表达的变化。结果：C5明显抑制CAL27、HN13和HN30细胞的生长，且与剂量和时间呈正相关。IC50值比GA对照组明显降低。应用5.0 μmol/L高浓度的C5，处理CAL27细胞系组24h、48h和72 h，细胞凋亡比例分别为52.19%、65.24%和84.53%。C5引起明显的、剂量依赖性的Bcl-2表达减少和Bax表达增加。结论：C5可以体外通过上调Bax蛋白和下调Bcl-2蛋白表达诱导头颈部鳞状细胞癌细胞的凋亡。

Abstract: [Abstract] Objective：To observe the effect of gambogic acid (GA) derivative compound 5 (C5) on apoptosis in human head and neck squamous cell carcinoma (HNSCC) in vitro. Methods：In the present study, HNSCC cell lines CAL27, HN13 and HN30 were treated with C5 at different doses for different times. The cell viability was detected with MTT assay and the IC50 values were also determined by Logit method. Cell apoptotic rates induced by C5 in the three cell lines were observed by Annexin-V/PI doubling staining by the flow cytometry assay. The changes on the s of apoptosis-related proteins were detected by Western blot and fluorescent immunocytochemistry assay. Results：CAL27, HN13 and HN30 cell lines were dramatically inhibited in a dose- and time-dependent manner after incubation with C5. The IC50 values were less than GA. When CAL27 cells were incubated with high dose C5 of 5.0 μmol/L for 24h, 48h and 72 h, the apoptotic rates were about 52.19%, 65.24% and 84.53%, respectively. C5 decreased protein levels of Bcl-2 and increased protein levels of Bax in the CAL27, HN13 and HN30 cell lines. Conclusions：Altogether, our data demonstrated that C5 induced apoptosis of HNSCC cell lines with up-regulation of Bax and down-regulation of Bcl-2 in vitro.

李腾宇潘劲松王旭. 藤黄酸衍生物-5诱导人头颈部鳞状细胞癌细胞凋亡的体外研究[J]. , 2016, 7(1): 6-11.

参考文献

[1]Siegel R,Ward E,Brawley O,et al.Cancer statistics,2011: the impact of eliminating socioeconomic and racial disparities on premature cancer deaths[J].CA Cancer J Clin,2011,61(4):212-236.
[2]Athanasoulis CA,Kaufman JA,Halpern EF,et al.Inferior vena caval filters: review of a 26-2000 single-center clinical experience[J].Radiology,2000,216(1):54-66.
[3]Lee UL,Choi SW.The chemopreventive properties and therapeutic modulation of green tea polyphenols in oral squamous cell carcinoma[J].ISRN Oncol,2011,2011:403707-.
[4]Niture SK,Jaiswal AK.Nrf2 protein up-regulates antiapoptotic protein Bcl-2 and prevents cellular apoptosis[J].J Biol Chem,2012,287(13):9873-9886.
[5]Zou ZY,Wei J,Li XL,et al.Enhancement of anticancer efficacy of chemotherapeutics by gambogic Acid against gastric cancer cells[J].Cancer Biother Radiopharm,2012,27(5):299-306.
[6]Huang H,Chen D,Li S,et al.Gambogic acid enhances proteasome inhibitor-induced anticancer activity[J].Cancer Lett,2011,301(2):221-228.
[7]Guo QL,You QD,Wu ZQ,et al.General gambogic acids inhibited growth of human hepatoma SMMC-7721 cells in vitro and in nude mice[J].Acta Pharmacol Sin,2004,25(6):769-774.
[8]Han QB,Cheung S,Tai J,et al.Stability and cytotoxicity of gambogic acid and its derivative,gambogoic acid[J].Biol Pharm Bull,2005,28(12):2335-2337.
[9]Liu W,Guo QL,You QD,et al.Anticancer effect and apoptosis induction of gambogic acid in human gastric cancer line BGC-823[J].World J Gastroenterol,2005,11(24):3655-3659.
[10]Wu ZQ,Guo QL,You QD,et al.Gambogic acid inhibits proliferation of human lung carcinoma SPC-A1 cells in vivo and in vitro and represses telomerase activity and telomerase reverse transcriptase mRNA expression in the cells[J].Biol Pharm Bull,2004,27(11):1769-1774.
[11]Wang K,Tang Y,Sun M,et al.The mechanism of neogambogic acid-induced apoptosis in human MCF-7 cells[J].Acta Biochim Biophys Sin,2011,43(9):698-702.
[12]Jégu J,Binder-Foucard F,Borel C,et al.Trends over three decades of the risk of second primary cancer among patients with head and neck cancer[J].Oral Oncol,2013,49(1):9-14.
[13]Lee JJ,Wu X,Hildebrandt MA,et al.Global assessment of genetic variation influencing response to retinoid chemoprevention in head and neck cancer patients[J].Cancer Prev Res,2011,4(2):185-193.
[14]Katada C,Muto M,Nakayama M,et al.Risk of superficial squamous cell carcinoma developing in the head and neck region in patients with esophageal squamous cell carcinoma[J].Laryngoscope,2012,122(6):1291-1296.
[15]Portt L,Norman G,Clapp C,et al.Anti-apoptosis and cell survival: a review[J].Biochim Biophys Acta,2011,1813(1):238-259.
[16]Galluzzi L,Vitale I,Abrams JM,et al.Molecular definitions of cell death subroutines: recommendations of the Nomenclature Committee on Cell Death 2012[J].Cell Death Differ,2012,19(1):107-120.
[17]Fesik SW.Promoting apoptosis as a strategy for cancer drug discovery[J].Nat Rev Cancer,2005,5(11):876-885.
[18]Zhang YH,Peng HY,Xia GH,et al.Anticancer effect of two diterpenoid compounds isolated from Annona glabra Linn[J].Acta Pharmacol Sin,2004,25(7):937-942.
[19]Zou Z,Xie L,Wei J,et al.Synergistic anti-proliferative effects of gambogic acid with docetaxel in gastrointestinal cancer cell lines[J].BMC Complement Altern Med,2012,12:58-.
[20] Chen HB,Zhou LZ,Mei L,et al.Gambogenic acid-induced time- and dose-dependent growth inhibition and apoptosis involving Akt pathway inactivation in U251 glioblastoma cells [J].J Nat Med,2012,66(1):62-69.
[21]Chen T,Zhang RH,He SC,et al.Synthesis and antiangiogenic activity of novel gambogic acid derivatives [J].Molecules,2012,17(6):6249-6268.
[22] Yang J,Ding L,Hu L,et al.Metabolism of gambogic acid in rats: a rare intestinal metabolic pathway responsible for its final disposition[J].Drug Metab Dispos,2011,39(4):617-626.
[23]Acoca S,Cui Q,Shore GC,et al.Molecular dynamics study of small molecule inhibitors of the Bcl-2 family[J].Proteins,2011,79(9):2624-2636.
[24]Wang SH,Martin SM,Harris PS,et al.Caspase inhibition blocks cell death and enhances mitophagy but fails to promote T-cell lymphoma[J].PLoS One,2011,6(5):e19786-.
[25]Croker BA,O'Donnell JA,Nowell CJ,et al.Fas-mediated neutrophil apoptosis is accelerated by Bid,Bak,and Bax and inhibited by Bcl-2 and Mcl-1[J].Proc Natl Acad Sci U S A,2011,108(32):13135-13140.
[26]Silva RD,Manon S,Gon?alves J,et al.The importance of humanized yeast to better understand the role of bcl-2 family in apoptosis: finding of novel therapeutic opportunities[J].Curr Pharm Des,2011,17(3):246-255.
[27] Laulier C,Lopez BS.The secret life of Bcl-2: apoptosis-independent inhibition of DNA repair by Bcl-2 family members[J].Mutat Res,2012,751(2):247-257.
[28]Yang D,Liu X,Zhang R,et al.Increased apoptosis and different regulation of pro-apoptosis protein bax and anti-apoptosis protein bcl-2 in the olfactory bulb of a rat model of depression[J].Neurosci Lett,2011,504(1):18-22.
[29]Gogada R,Prabhu V,Amadori M,et al.Resveratrol induces p53-independent,X-linked inhibitor of apoptosis protein (XIAP)-mediated Bax protein oligomerization on mitochondria to initiate cytochrome c release and caspase activation[J].J Biol Chem,2011,286(33):28749-28760.
[30]Rossé T,Olivier R,Monney L,et al.Bcl-2 prolongs cell survival after Bax-induced release of cytochromec[J].Nature,1998,391(6666):496-499.