藤黄酸对顺铂诱导A549/DDP细胞凋亡的增敏作用
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摘要
目的:探讨藤黄酸(GA)是否能作为肺癌化疗中顺铂(DDP)的增敏剂。方法:采用细胞增殖试验和等辐射分析法研究DDP联合GA对非小细胞肺癌耐药株A549/DDP的抑制作用,并测定其对B淋巴细胞瘤-2(Bcl-2)和P-糖蛋白(P-gp)表达的影响。结果:GA可显著增强A549/DDP细胞对DDP介导的细胞凋亡的敏感性,显著增加DDP诱导细胞的早期凋亡(20.58±5.68)%、晚期凋亡(17.47±4.05)%和G_2/M期停滞(44.13±3.78)%。GA与DDP联合组中,P-gp及抗凋亡蛋白Bcl-2被显著抑制(P=0.006及P=0.007),促凋亡蛋白Bax显著增加(P=0.006)。此外,联合治疗也抑制了p53蛋白(P<0.01)的表达。结论:GA通过增加DDP在细胞内积聚和增强DDP介导的DDP耐药肺癌细胞凋亡来增强DDP的作用,GA可以作为DDP诱导A549/DDP细胞凋亡的良好增敏剂,但其潜在的机制尚待进一步探明。
Objective: To determine whether gambogic acid(GA) is a sensitizer for cisplatin(DDP) in chemotherapy of lung cancer. Methods: Cell proliferation test and other radiation analysis were used to study the inhibitory effect of DDP combined with GA on A549/DDP, and to determine its effect on the expression of B-cell lymphoma-2(Bcl-2) and p-glycoprotein(P-gp). Results: GA could significantly enhance the sensitivity of A549/DDP cells to apoptosis induced by DDP. It significantly increased the early apoptosis(20.58±5.68)%, late apoptosis(17.47±4.05)% and G_2/M-stage stagnation(44.13±3.78)% induced by cisplatin. In the combined group of GA and DDP, the P-gp and anti-apoptotic protein Bcl-2 were significantly inhibited(P=0.006), and pro-apoptotic protein Bcl-2 associated X protein(Bax)(P=0.006) increased significantly. In addition, combined treatment also inhibited the expression of p53 protein(P<0.01). Conclusion: GA can enhance the effect of cisplatin by increasing the accumulation of DDP in cells and enhancing the apoptosis induced by cisplatin in cisplatin-resistant no small cell lung cancer cells,and GA is a good sensitizer for the apoptosis of A549/DDP cells induced by DDP. But its potential mechanisms need to be further elaborated.
Objective: To determine whether gambogic acid(GA) is a sensitizer for cisplatin(DDP) in chemotherapy of lung cancer. Methods: Cell proliferation test and other radiation analysis were used to study the inhibitory effect of DDP combined with GA on A549/DDP, and to determine its effect on the expression of B-cell lymphoma-2(Bcl-2) and p-glycoprotein(P-gp). Results: GA could significantly enhance the sensitivity of A549/DDP cells to apoptosis induced by DDP. It significantly increased the early apoptosis(20.58±5.68)%, late apoptosis(17.47±4.05)% and G_2/M-stage stagnation(44.13±3.78)% induced by cisplatin. In the combined group of GA and DDP, the P-gp and anti-apoptotic protein Bcl-2 were significantly inhibited(P=0.006), and pro-apoptotic protein Bcl-2 associated X protein(Bax)(P=0.006) increased significantly. In addition, combined treatment also inhibited the expression of p53 protein(P<0.01). Conclusion: GA can enhance the effect of cisplatin by increasing the accumulation of DDP in cells and enhancing the apoptosis induced by cisplatin in cisplatin-resistant no small cell lung cancer cells,and GA is a good sensitizer for the apoptosis of A549/DDP cells induced by DDP. But its potential mechanisms need to be further elaborated.
引文
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[19]CHEE J L,SAIDIN S,LANE D P,et al.Wild- type and mutant p53 mediate cisplatin resistance through interaction and inhibition of active caspase- 9[J].Cell Cycle,2013,12(2):278- 288.
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[22]郭晓彤,魏优蕾,雷加吉,等.藤黄酸对非小细胞肺癌A549细胞凋亡及Bcl- 2、Bax、P53基因表达的影响[J].中国老年学杂志,2018,38(4):911- 914.
[2] AZZOLI C G,TEMIN S,ALIFF T,et al.2011 Focused Update of 2009 American Society of Clinical Oncology Clinical Practice Guideline update on chemotherapy for stage Ⅳ non- small- cell lung cancer[J].J Clin Oncol,2011,29(28):3825- 3831.
[3] ETTINGER D S,WOOD D E,AKERLEY W,et al.Non- small cell lung cancer,version 6.2015[J].J Natl Compr Canc Netw,2015,13(5):515- 524.
[4] AZZOLI C G,TEMIN S,GIACCONE G.2011 Focused Update of 2009 American Society of Clinical Oncology Clinical Practice Guideline update on chemotherapy for stage Ⅳ non- small- cell lung cancer[J].J Oncol Pract,2012,8(1):63- 66.
[5] ZHAO T,WANG H J,ZHAO W W,et al.Gambogic acid improves non- small cell lung cancer progression by inhibition of mTOR signaling pathway[J].Kaohsiung J Med Sci,2017,33(11):543- 549.
[6] ZHANG H Z,KASIBHATLA S,WANG Y,et al.Discovery,characterization and SAR of gambogic acid as a potent apoptosis inducer by a HTS assay[J].Bioorg Med Chem,2004,12(2):309- 317.
[7] CHEN Q,YU Q,LIU Y,et al.Multifunctional selenium nanoparticles:chiral selectivity of delivering MDR- siRNA for reversal of multidrug resistance and real- time biofluorescence imaging[J].Nanomedicine,2015,11(7):1773- 1784.
[8] YANG L,SHI T,LIU F,et al.REV3L,a promising target in regulating the chemosensitivity of cervical cancer cells[J].PLoS One,2015,10(3):334- 352.
[9] MIRANDA H F,PUIG M M,PRIETO J C,et al.Synergism between paracetamol and nonsteroidal anti- inflammatory drugs in experimental acute pain[J].Pain,2006,121(1- 2):22- 28.
[10]HUANG J,ZHU X,WANG H,et al.Role of gambogic acid and NaI131 in A549/DDP cells[J].Oncol Lett,2017,13(1):37- 44.
[11]CHEN P M,CHENG Y W,WU T C,et al.MnSOD overexpression confers cisplatin resistance in lung adenocarcinoma via the NF- κB/Snail/Bcl- 2 pathway[J].Free Radic Biol Med,2015,79:127- 137.
[12]WU D W,WU T C,WU J Y,et al.Phosphorylation of paxillin confers cisplatin resistance in non- small cell lung cancer via activating ERK- mediated Bcl- 2 expression[J].Oncogene,2014,33(35):4385- 4395.
[13]NISHIOKA T,LUO L Y,SHEN L,et al.Nicotine increases the resistance of lung cancer cells to cisplatin through enhancing Bcl- 2 stability[J].Br J Cancer,2014,110(7):1785- 1792.
[14]LAN D,WANG L,HE R,et al.Exogenous glutathione contributes to cisplatin resistance in lung cancer A549 cells[J].Am J Transl Res,2018,10(5):1295- 1309.
[15]CHEN Y L,YANG T Y,CHEN K C,et al.Hypoxia can impair doxorubicin resistance of non- small cell lung cancer cells by inhibiting MRP1 and P- gp expression and boosting the chemosensitizing effects of MRP1 and P- gp blockers[J].Cell Oncol(Dordr),2016,39(5):411- 433.
[16]LE?O M,GOMES S,PEDRAZA- CHAVERRI J,et al.A mangostin and gambogic acid as potential inhibitors of the p53- MDM2 interaction revealed by a yeast approach[J].J Nat Prod,2013,76(4):774- 778.
[17]KOSTER R,TIMMER- BOSSCHA H,BISCHOFF R,et al.Disruption of the MDM2- p53 interaction strongly potentiates p53- dependent apoptosis in cisplatin- resistant human testicular carcinoma cells via the Fas/FasL pathway[J].Cell Death Dis,2011,2:148- 159.
[18]TANG X,HU G,XU C,et al.HZ08 reverse the aneuploidy- induced cisplatin- resistance in Gastric cancer by modulating the p53 pathway[J].Eur J Pharmacol,2013,720(1- 3):84- 97.
[19]CHEE J L,SAIDIN S,LANE D P,et al.Wild- type and mutant p53 mediate cisplatin resistance through interaction and inhibition of active caspase- 9[J].Cell Cycle,2013,12(2):278- 288.
[20]SARIN N,ENGEL F,KALAYDA G V,et al.Cisplatin resistance in non- small cell lung cancer cells is associated with an abrogation of cisplatin- induced G2/M cell cycle arrest[J].PLoS One,2017,12(7):81- 99.
[21]GADHIKAR M A,SCIUTO M R,ALVES M V,et al.Chk1/2 inhibition overcomes the cisplatin resistance of head and neck cancer cells secondary to the loss of functional p53[J].Mol Cancer Ther,2013,12(9):1860- 1873.
[22]郭晓彤,魏优蕾,雷加吉,等.藤黄酸对非小细胞肺癌A549细胞凋亡及Bcl- 2、Bax、P53基因表达的影响[J].中国老年学杂志,2018,38(4):911- 914.