PPARγ配体激动剂罗格列酮对甲状腺乳头状癌BCPAP细胞增殖、凋亡的影响及机制
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摘要
目的观察PPARγ配体激动剂罗格列酮(ROZ)对甲状腺乳头状癌(PTC) BCPAP细胞增殖、凋亡的影响,并探讨其机制。方法将细胞分别置于ROZ终浓度为0、20、40μmol/L培养基中,MTT比色法检测作用24、48、72 h时细胞增殖的光密度(OD)值。将细胞分为4组:A组不处理,B组加入终浓度为40μmol/L的ROZ处理,C组加入终浓度为1μmol/m L第10号染色体缺失的磷酸酶和张力蛋白同源基因(PTEN)特异性抑制剂bp V(phen)处理,D组先加入终浓度为1μmol/m L bp V(phen)处理12 h后再加入终浓度为40μmol/L的ROZ处理;各组处理48 h后,采用流式细胞仪测算细胞凋亡率,Western blotting法检测细胞中Bcl-2、Bax蛋白及PTEN、蛋白激酶B(AKT)、磷酸化蛋白激酶B(p-AKT)蛋白。结果随着ROZ浓度增加和作用时间的延长,BCPAP细胞增殖的OD值逐渐降低(P均<0. 01)。B组细胞凋亡率高于A、C、D组(P均<0. 01),A、C、D组间细胞凋亡率差异均无统计学意义(P均> 0. 05)。与A、C、D组比较,B组细胞Bax、PTEN蛋白相对表达量增加,而Bcl-2、p-AKT蛋白相对表达量减少(P均<0. 01),A、C、D组间细胞Bcl-2、Bax、PTEN、p-AKT、AKT蛋白相对表达量差异均无统计学意义(P均> 0. 05)。结论罗格列酮可能是通过PTEN/AKT通路来调节Bcl-2/Bax表达比例,从而抑制肿瘤细胞增殖并诱导细胞发生凋亡。
Objective To explore the effects and mechanism of PPARγ ligand agonist rosiglitazone( ROZ) on the proliferation and apoptosis of papillary thyroid carcinoma( PTC) BCPAP cells in vitro. Methods The cell proliferation optical density( OD) values were determined by MTT assay at 24,48 and 72 h after the cells were cultured in medium with a final ROZ concentration of 0,20,40 mol/L,respectively. Then we divided them into four groups,group A was not treated,group B was treated by ROZ with a final concentration of 40 μmol/L,group C was treated by 1 μmol/m L phosphatase and tensin homolog deleted on chromosome ten( PTEN) specific inhibitor bp V( phen),and group D was treated with 1μmol/m L bp V( phen) for 12 h,followed by 40 μmol/L ROZ. After treatment for 48 h in each group,the apoptosis rate was measured by flow cytometry,and the Bcl-2 and Bax proteins,and PTEN,protein kinase B( AKT),and phosphorylated protein kinase B( p-AKT) proteins were detected by Western blotting. Results With the increase of ROZ concentration and the extension of action time,the OD value of BCPAP cells gradually decreased( all P < 0. 01). The apoptosis rate of group B was higher than those of groups A,C and D( all P < 0. 01),and no significant difference was found in the apoptosis rates between groups A,C and D( all P > 0. 05). Compared with groups A,C and D,the relative expression levels of Bax and PTEN proteins in cells of group B increased,while the relative expression levels of Bcl-2 and p-AKT proteins decreased( all P < 0. 01). No significant differences were found in the relative expression levels of Bcl-2,Bax,PTEN,pAKT and AKT among groups A,C and D( all P > 0. 05). Conclusion ROZ may regulate the Bcl-2/Bax expression ratio through the PTEN/AKT pathway,thereby inhibiting the proliferation of tumor cells and inducing apoptosis.
Objective To explore the effects and mechanism of PPARγ ligand agonist rosiglitazone( ROZ) on the proliferation and apoptosis of papillary thyroid carcinoma( PTC) BCPAP cells in vitro. Methods The cell proliferation optical density( OD) values were determined by MTT assay at 24,48 and 72 h after the cells were cultured in medium with a final ROZ concentration of 0,20,40 mol/L,respectively. Then we divided them into four groups,group A was not treated,group B was treated by ROZ with a final concentration of 40 μmol/L,group C was treated by 1 μmol/m L phosphatase and tensin homolog deleted on chromosome ten( PTEN) specific inhibitor bp V( phen),and group D was treated with 1μmol/m L bp V( phen) for 12 h,followed by 40 μmol/L ROZ. After treatment for 48 h in each group,the apoptosis rate was measured by flow cytometry,and the Bcl-2 and Bax proteins,and PTEN,protein kinase B( AKT),and phosphorylated protein kinase B( p-AKT) proteins were detected by Western blotting. Results With the increase of ROZ concentration and the extension of action time,the OD value of BCPAP cells gradually decreased( all P < 0. 01). The apoptosis rate of group B was higher than those of groups A,C and D( all P < 0. 01),and no significant difference was found in the apoptosis rates between groups A,C and D( all P > 0. 05). Compared with groups A,C and D,the relative expression levels of Bax and PTEN proteins in cells of group B increased,while the relative expression levels of Bcl-2 and p-AKT proteins decreased( all P < 0. 01). No significant differences were found in the relative expression levels of Bcl-2,Bax,PTEN,pAKT and AKT among groups A,C and D( all P > 0. 05). Conclusion ROZ may regulate the Bcl-2/Bax expression ratio through the PTEN/AKT pathway,thereby inhibiting the proliferation of tumor cells and inducing apoptosis.
引文
[1]马振海,田晓峰,赵永福,等.甲状腺乳头状癌术后复发影响因素分析[J].中国普外基础与临床杂志,2013,20(12):1410-1413.
[2]翟宝伟,高庆军,赵代伟.甲状腺微小乳头状癌颈部淋巴结转移的危险因素分析[J].中国普通外科杂志,2016,25(11):1573-1579.
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[4]Chai BK,Lau YS,Loong BJ,et al.Co-administration of conjugated linoleic acid and rosiglitazone increases atherogenic co-efficicient and alters isoprenaline-induced vasodilatation in rats fed high fat diet[J].Physiol Res,2018,67(5):729-740.
[5]包文影,于丽波,孙文洲,等.罗格列酮促进卵巢癌细胞凋亡的体内实验研究[J].中国肿瘤,2014,23(8):699-703.
[6]戴丹,查震球,陈叶纪,等.安徽省2010~2013年女性甲状腺癌发病趋势分析[J].现代预防医学,2018,45(1):51-53.
[7]尚培中,柳勇,南润玲.甲状腺乳头状癌的诊断与治疗[J].中国现代普通外科进展,2015,18(10):833-836.
[8]王莎莎,林岩松,梁军,等.甲状腺乳头状癌首次治疗术式对预后影响的分析[J].中华肿瘤防治杂志,2012,19(10):761-765.
[9]Daniel FI,Lima LD,Grando LJ,et al.Salivary evaluation in radioactive I(131)treated patients with thyroid carcinoma[J].Acta Odontol Scand,2018,76(2):148-152.
[10]Meng Q,Fang W,Long X,et al.Sialoendoscopy combined with an internal stent and postoperative massage as a comprehensive treatment of delayed I(131)-induced parotitis[J].Br J Oral Maxillofac Surg,2017,55(7):674-678.
[11]Krawczyk-Rusiecka K,Wojciechowska-Durczynska K,CyniakMagierska A,et al.COX-2 expression in papillary thyroid carcinoma(PTC)in cytological material obtained by fine needle aspiration biopsy(FNAB)[J].Thyroid Res,2011,4(1):3.
[12]Moschos MM,Chatziralli I,Brouzas D,et al.BAX and BCL2gene polymorphisms in rhegmatogenous retinal detachment[J].Ophthalmic Res,2017,58(4):227-230.
[13]Chen Y,Zhou R,Yi Z,et al.Porphyromonas gingivalis induced inflammatory responses and promoted apoptosis in lung epithelial cells infected with H1N1 via the Bcl2/Bax/Caspase3 signaling pathway[J].Mol Med Rep,2018,18(1):97-104.
[14]Zadi HM,Vatanmakanian M,Abdolalizadeh J,et al.Apoptotic effect of resveratrol on human T-ALL cell line CCRF-CEM is unlikely exerted through alteration of BAX and BCL2 promoter methylation[J].J Cell Biochem,2018,119(12):10033-10040.
[15]Xie L,Wu Y,Fan Z,et al.Astragalus polysaccharide protects human cardiac microvascular endothelial cells from hypoxia/reoxygenation injury:the role of PI3K/AKT,Bax/Bcl-2 and caspase-3[J].Mol Med Rep,2016,14(1):904-910.
[16]Li J,Gong X,Jiang R,et al.Fisetin inhibited growth and metastasis of triple-negative breast cancer by reversing epithelial-to-mesenchymal transition via PTEN/Akt/GSK3beta signal pathway[J].Front Pharmacol,2018,9:772.
[17]Zhang GL,Song JL,Ji CL,et al.Zearalenone exposure enhanced the expression of tumorigenesis genes in donkey granulosa cells via the PTEN/PI3K/AKT signaling pathway[J].Front Genet,2018,9:293.
[18]Song Z,Han X,Shen L,et al.PTEN silencing enhances neuronal proliferation and differentiation by activating PI3K/Akt/GSK3beta pathway in vitro[J].Exp Cell Res,2018,363(2):179-187.
[19]Jin H,Sun Y,Wang S,et al.Matrine activates PTEN to induce growth inhibition and apoptosis in V600EBRAF harboring melanoma cells[J].Int J Mol Sci,2013,14(8):16040-16057.
[20]Cheng D,Gao H,Li W.Long-term risk of rosiglitazone on cardiovascular events:a systematic review and meta-analysis[J].Endokrynol Pol,2018,69(4):381-394.
[21]Cheng WY,Huynh H,Chen P,et al.Macrophage PPARgamma inhibits Gpr132 to mediate the anti-tumor effects of rosiglitazone[J].Elife,2016,5:e18501.
[22]An Z,Yu JR,Park WY.Rosiglitazone enhances radiosensitivity by inhibiting repair of DNA damage in cervical cancer cells[J].Radiat Environ Biophys,2017,56(1):89-98.
[23]张琪,杨光华,葛志鹏,等.罗格列酮对HT29、HCT116结肠癌细胞凋亡影响及机制探究[J].中国比较医学杂志,2017,27(12):56-60.
[24]Chen Q,Yue F,Li W,et al.Potassium bisperoxo(1,10-phenanthroline)oxovanadate bpV(phen)induces apoptosis and pyroptosis and disrupts the P62-HDAC6 protein interaction to suppress the acetylated microtubule-dependent degradation of autophagosomes[J].J Biol Chem,2015,290(43):26051-26058.
[2]翟宝伟,高庆军,赵代伟.甲状腺微小乳头状癌颈部淋巴结转移的危险因素分析[J].中国普通外科杂志,2016,25(11):1573-1579.
[3]Peters AL.50 years ago in the journal of pediatrics:the diagnosis of complete extrahepatic obstruction by rose bengal131I[J].J Pediatr,2017,180:162.
[4]Chai BK,Lau YS,Loong BJ,et al.Co-administration of conjugated linoleic acid and rosiglitazone increases atherogenic co-efficicient and alters isoprenaline-induced vasodilatation in rats fed high fat diet[J].Physiol Res,2018,67(5):729-740.
[5]包文影,于丽波,孙文洲,等.罗格列酮促进卵巢癌细胞凋亡的体内实验研究[J].中国肿瘤,2014,23(8):699-703.
[6]戴丹,查震球,陈叶纪,等.安徽省2010~2013年女性甲状腺癌发病趋势分析[J].现代预防医学,2018,45(1):51-53.
[7]尚培中,柳勇,南润玲.甲状腺乳头状癌的诊断与治疗[J].中国现代普通外科进展,2015,18(10):833-836.
[8]王莎莎,林岩松,梁军,等.甲状腺乳头状癌首次治疗术式对预后影响的分析[J].中华肿瘤防治杂志,2012,19(10):761-765.
[9]Daniel FI,Lima LD,Grando LJ,et al.Salivary evaluation in radioactive I(131)treated patients with thyroid carcinoma[J].Acta Odontol Scand,2018,76(2):148-152.
[10]Meng Q,Fang W,Long X,et al.Sialoendoscopy combined with an internal stent and postoperative massage as a comprehensive treatment of delayed I(131)-induced parotitis[J].Br J Oral Maxillofac Surg,2017,55(7):674-678.
[11]Krawczyk-Rusiecka K,Wojciechowska-Durczynska K,CyniakMagierska A,et al.COX-2 expression in papillary thyroid carcinoma(PTC)in cytological material obtained by fine needle aspiration biopsy(FNAB)[J].Thyroid Res,2011,4(1):3.
[12]Moschos MM,Chatziralli I,Brouzas D,et al.BAX and BCL2gene polymorphisms in rhegmatogenous retinal detachment[J].Ophthalmic Res,2017,58(4):227-230.
[13]Chen Y,Zhou R,Yi Z,et al.Porphyromonas gingivalis induced inflammatory responses and promoted apoptosis in lung epithelial cells infected with H1N1 via the Bcl2/Bax/Caspase3 signaling pathway[J].Mol Med Rep,2018,18(1):97-104.
[14]Zadi HM,Vatanmakanian M,Abdolalizadeh J,et al.Apoptotic effect of resveratrol on human T-ALL cell line CCRF-CEM is unlikely exerted through alteration of BAX and BCL2 promoter methylation[J].J Cell Biochem,2018,119(12):10033-10040.
[15]Xie L,Wu Y,Fan Z,et al.Astragalus polysaccharide protects human cardiac microvascular endothelial cells from hypoxia/reoxygenation injury:the role of PI3K/AKT,Bax/Bcl-2 and caspase-3[J].Mol Med Rep,2016,14(1):904-910.
[16]Li J,Gong X,Jiang R,et al.Fisetin inhibited growth and metastasis of triple-negative breast cancer by reversing epithelial-to-mesenchymal transition via PTEN/Akt/GSK3beta signal pathway[J].Front Pharmacol,2018,9:772.
[17]Zhang GL,Song JL,Ji CL,et al.Zearalenone exposure enhanced the expression of tumorigenesis genes in donkey granulosa cells via the PTEN/PI3K/AKT signaling pathway[J].Front Genet,2018,9:293.
[18]Song Z,Han X,Shen L,et al.PTEN silencing enhances neuronal proliferation and differentiation by activating PI3K/Akt/GSK3beta pathway in vitro[J].Exp Cell Res,2018,363(2):179-187.
[19]Jin H,Sun Y,Wang S,et al.Matrine activates PTEN to induce growth inhibition and apoptosis in V600EBRAF harboring melanoma cells[J].Int J Mol Sci,2013,14(8):16040-16057.
[20]Cheng D,Gao H,Li W.Long-term risk of rosiglitazone on cardiovascular events:a systematic review and meta-analysis[J].Endokrynol Pol,2018,69(4):381-394.
[21]Cheng WY,Huynh H,Chen P,et al.Macrophage PPARgamma inhibits Gpr132 to mediate the anti-tumor effects of rosiglitazone[J].Elife,2016,5:e18501.
[22]An Z,Yu JR,Park WY.Rosiglitazone enhances radiosensitivity by inhibiting repair of DNA damage in cervical cancer cells[J].Radiat Environ Biophys,2017,56(1):89-98.
[23]张琪,杨光华,葛志鹏,等.罗格列酮对HT29、HCT116结肠癌细胞凋亡影响及机制探究[J].中国比较医学杂志,2017,27(12):56-60.
[24]Chen Q,Yue F,Li W,et al.Potassium bisperoxo(1,10-phenanthroline)oxovanadate bpV(phen)induces apoptosis and pyroptosis and disrupts the P62-HDAC6 protein interaction to suppress the acetylated microtubule-dependent degradation of autophagosomes[J].J Biol Chem,2015,290(43):26051-26058.
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