Bcl-2抑制剂ABT-263对鼻咽癌细胞5-8F增殖和凋亡的影响
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摘要
目的研究B细胞淋巴瘤/白血病2(Bcl-2)抑制剂ABT-263对鼻咽癌细胞5-8F增殖和凋亡的影响。方法采用细胞计数试剂盒8检测不同浓度下Bcl-2抑制剂ABT-263对鼻咽癌细胞5-8F增殖的影响; Annexin V-异硫氰酸荧光素/碘化丙啶双染法检测不同浓度ABT-263对鼻咽癌细胞5-8F凋亡的影响;应用免疫印迹法检测抑制剂ABT-263对Bcl-2、Bcl-xL、髓细胞白血病1(Mcl-1)及佛波醇-12-肉豆蔻酸-13-乙酸酯诱导的蛋白质1(又称Noxa)等蛋白表达水平的影响。结果 Bcl-2抑制剂ABT-263处理细胞后,0. 125、0. 25、0. 5、1、2、4、8μmol/L组吸光值低于0μmol/L组(0. 696±0. 020,0. 527±0. 015,0. 465±0. 022,0. 323±0. 010,0. 133±0. 004,0. 057±0. 002,0. 028±0. 002比0. 856±0. 017)(P <0. 05)。在0. 5μmol/L浓度下诱导约6%鼻咽癌细胞的凋亡,其诱导凋亡的机制可能主要是通过正调控促凋亡蛋白Noxa完成的,在1μmol/L ABT-263的作用下能够引起鼻咽癌细胞中Noxa近7倍左右的升高。结论 ABT-263可以通过上调促凋亡蛋白Noxa促进鼻咽癌细胞的凋亡,具有应用于鼻咽癌治疗的潜力。
Objective To investigate the effect of B-cell lymphoma/leukemia-2( Bcl-2) inhibitor ABT-263 on proliferation and apoptosis of nasopharyngeal carcinoma( NPC) cell line 5-8 F. Methods Cell counting Kit-8 was used to detect the effect of Bcl-2 inhibitor ABT-263 on the proliferation of nasopharyngeal carcinoma cells 5-8 F. The apoptosis effect of ABT-263 on 5-8 F was detected by Annexin V-Fluoresceine isothiocyanate/propidium iodide double staining. The Western Blot assay was used to measure the effect of inhibitor ABT-263 on the expression of Bcl-2,Bcl-extra large( Bcl-xL),myeloid cell leukemia-1( Mcl-1),phorbol-12-myristate-13-acetate-induced protein 1( Noxa) and other Bcl-2 family proteins. Results After treatment of the cells with Bcl-2 inhibitor ABT-263,the absorbance values of 0. 125,0. 25,0. 5,1,2,4,8 μmol/L groups were lower than that of the 0 μmol/L group( 0. 696 ± 0. 020,0. 527 ± 0. 015,0. 465 ± 0. 022,0. 323 ± 0. 010,0. 133 ± 0. 004,0. 057 ± 0. 002,0. 028 ± 0. 002 vs 0. 856 ± 0. 017)( P < 0. 05),0. 5 μmol/L ABT-263 can induce about apoptosis of about 6% of the NPC cells. The mechanism may be through the positive regulation of apoptosispromoting protein Noxa. Under the effect of 1 μmol/L ABT-263,Noxa in NPC cells increased nearly 7 times. Conclusion ABT-263 can promote NPC cell apoptosis through up-regulating Noxa expression,and is potentially applicable in NPC treatment.
Objective To investigate the effect of B-cell lymphoma/leukemia-2( Bcl-2) inhibitor ABT-263 on proliferation and apoptosis of nasopharyngeal carcinoma( NPC) cell line 5-8 F. Methods Cell counting Kit-8 was used to detect the effect of Bcl-2 inhibitor ABT-263 on the proliferation of nasopharyngeal carcinoma cells 5-8 F. The apoptosis effect of ABT-263 on 5-8 F was detected by Annexin V-Fluoresceine isothiocyanate/propidium iodide double staining. The Western Blot assay was used to measure the effect of inhibitor ABT-263 on the expression of Bcl-2,Bcl-extra large( Bcl-xL),myeloid cell leukemia-1( Mcl-1),phorbol-12-myristate-13-acetate-induced protein 1( Noxa) and other Bcl-2 family proteins. Results After treatment of the cells with Bcl-2 inhibitor ABT-263,the absorbance values of 0. 125,0. 25,0. 5,1,2,4,8 μmol/L groups were lower than that of the 0 μmol/L group( 0. 696 ± 0. 020,0. 527 ± 0. 015,0. 465 ± 0. 022,0. 323 ± 0. 010,0. 133 ± 0. 004,0. 057 ± 0. 002,0. 028 ± 0. 002 vs 0. 856 ± 0. 017)( P < 0. 05),0. 5 μmol/L ABT-263 can induce about apoptosis of about 6% of the NPC cells. The mechanism may be through the positive regulation of apoptosispromoting protein Noxa. Under the effect of 1 μmol/L ABT-263,Noxa in NPC cells increased nearly 7 times. Conclusion ABT-263 can promote NPC cell apoptosis through up-regulating Noxa expression,and is potentially applicable in NPC treatment.
引文
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[18] Zhen Y,Liu Z,Yang H,et al. Tumor suppressor PDCD4 modulates miR-184-mediated direct suppression of C-MYC and BCL2blocking cell growth and survival in nasopharyngeal carcinoma[J].Cell Death Dis,2013,4:e872.
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[22] Kotschy A,Szlavik Z,Murray J,et al. The MCL1 inhibitor S63845is tolerable and effective in diverse cancer models[J]. Nature,2016,538(7626):477-482.
[23] Tse C,Shoemaker AR,Adickes J,et al. ABT-263:A potent and orally bioavailable Bcl-2 family inhibitor[J]. Cancer Res,2008,68(9):3421-3428.
[24] Wang Z,Zhao Q,Zhang D,et al. Essential roles for platelets during neutrophil-dependent or lymphocyte-mediated defense against bacterial pathogens[J]. Blood Coagul Fibrinolysis,2016,27(6):667-672.
[2] Bose P,Gandhi V,Konopleva M. Pathways and mechanisms of venetoclax resistance[J]. Leuk Lymphoma,2017,58(9):1-17.
[3] Souers AJ,Leverson JD,Boghaert ER,et al. ABT-199,a potent and selective Bcl-2 inhibitor,achieves antitumor activity while sparing platelets[J]. Nat Med,2013,19(2):202-208.
[4] Lu QL,Elia G,Lucas S,et al. Bcl-2 proto-oncogene expression in Epstein-Barr-virus-associated nasopharyngeal carcinoma[J]. Int J Cancer,1993,53(1):29-35.
[5] Wendt MD. Discovery of ABT-263,a Bcl-family protein inhibitor:Observations on targeting a large protein-protein interaction[J].Expert Opin Drug Discov,2008,3(9):1123-1143.
[6] Tang LL,Chen WQ,Xue WQ,et al. Global trends in incidence and mortality of nasopharyngeal carcinoma[J]. Cancer Lett,2016,374(1):22-30.
[7] Lee AW,Poon YF,Foo W,et al. Retrospective analysis of 5037patients with nasopharyngeal carcinoma treated during 1976-1985:Overall survival and patterns of failure[J]. Int J Radiat Oncol Biol Phys,1992,23(2):261-270.
[8] Chua MLK,Wee JTS,Hui EP,et al. Nasopharyngeal carcinoma[J].Lancet,2016,387(10022):1012-1024.
[9] Liu MT,Chen MK,Huang CC,et al. Prognostic Value of Molecular Markers and Implication for Molecular Targeted Therapies in Nasopharyngeal Carcinoma:An Update in an Era of New Targeted Molecules Development[J]. World J Oncol,2015,6(1):243-261.
[10] Hassan M,Watari H,Abu Almaaty A,et al. Apoptosis and molecular targeting therapy in cancer[J]. Biomed Res Int,2014,2014:150845.
[11] Bianchi F,Pretto S,Tagliabue E,et al. Exploiting poly(I:C)to induce cancer cell apoptosis[J]. Cancer Biol Ther,2017,18(10):747-756.
[12] Mohamed MS,Bishr MK,Almutairi FM,et al. Inhibitors of apoptosis:Clinical implications in cancer[J]. Apoptosis,2017,22(12):1487-1509.
[13] Roberts AW,Davids MS,Pagel JM,et al. Targeting BCL2 with Venetoclax in Relapsed Chronic Lymphocytic Leukemia[J].N Engl J Med,2016,374(4):311-322.
[14] Valentin R,Grabow S,Davids MS. The rise of apoptosis:Targeting apoptosis in hematologic malignancies[J]. Blood,2018,132(12):1248-1264.
[15] Oliver CL,Bauer JA,Wolter KG,et al. In vitro effects of the BH3mimetic,(-)-gossypol,on head and neck squamous cell carcinoma cells[J]. Clin Cancer Res,2004,10(22):7757-7763.
[16] Merino D,Lok SW,Visvader JE,et al. Targeting BCL-2 to enhance vulnerability to therapy in estrogen receptor-positive breast cancer[J]. Oncogene,2016,35(15):1877-1887.
[17] Tsang CM,Tsao SW. The role of Epstein-Barr virus infection in the pathogenesis of nasopharyngeal carcinoma[J]. Virol Sin,2015,30(2):107-121.
[18] Zhen Y,Liu Z,Yang H,et al. Tumor suppressor PDCD4 modulates miR-184-mediated direct suppression of C-MYC and BCL2blocking cell growth and survival in nasopharyngeal carcinoma[J].Cell Death Dis,2013,4:e872.
[19] Hu ZY,Zhu XF,Zhong ZD,et al. ApoG2,a novel inhibitor of antiapoptotic Bcl-2 family proteins,induces apoptosis and suppresses tumor growth in nasopharyngeal carcinoma xenografts[J].Int J Cancer,2008,123(10):2418-2429.
[20] Wang YJ,Wang YY,Fan XQ,et al. ABT-199-mediated inhibition of Bcl-2 as a potential therapeutic strategy for nasopharyngeal carcinoma[J]. Biochem Biophys Res Commun,2018,503(3):1214-1220.
[21] Low SY,Tan BS,Choo HL,et al. Suppression of BCL-2 synergizes cisplatin sensitivity in nasopharyngeal carcinoma cells[J]. Cancer Lett,2012,314(2):166-175.
[22] Kotschy A,Szlavik Z,Murray J,et al. The MCL1 inhibitor S63845is tolerable and effective in diverse cancer models[J]. Nature,2016,538(7626):477-482.
[23] Tse C,Shoemaker AR,Adickes J,et al. ABT-263:A potent and orally bioavailable Bcl-2 family inhibitor[J]. Cancer Res,2008,68(9):3421-3428.
[24] Wang Z,Zhao Q,Zhang D,et al. Essential roles for platelets during neutrophil-dependent or lymphocyte-mediated defense against bacterial pathogens[J]. Blood Coagul Fibrinolysis,2016,27(6):667-672.