Establishment and characterization of human osteosarcoma cells resistant to pyropheophorbide-α methyl ester-mediated photodynamic therapy.
Establishment and characterization of human osteosarcoma cells resistant to pyropheophorbide-α methyl ester-mediated photodynamic therapy.
复制标题
对焦脱镁叶绿酸-α甲酯介导的光动力疗法具有抗性的人骨肉瘤细胞的建立和表征
DOI：
10.3892/ijo.2017.4136
复制
发表时间：
2017-11
影响因子：
5.2
通讯作者：
Deng Q
中科院分区：
文献类型：
--
作者：
Tao Y;Ou Y;Yin H;Chen Y;Zhong S;Gao Y;Zhao Z;He B;Huang Q;Deng Q
本研究旨在建立和鉴定对焦脱镁叶绿酸-α甲酯介导的光动力疗法耐药的新的人骨肉瘤细胞系。从人骨肉瘤细胞系MG63和HOS中分离出MPPA-PDT耐药细胞，最终获得MG63/PDT和HOS/PDT两个耐药细胞系。采用细胞计数试剂盒-8(CCK-8)法检测MG63、MG63/PDT、HOS和HOS/PDT细胞对MPPA-PDT、顺铂(CDDP)耐药和增殖的影响。DCFH-DA染色分析细胞内ROS。比较亲本细胞和抗性细胞的集落形成、侵袭和迁移情况。流式细胞仪检测细胞周期分布、凋亡率和CD133+细胞比例。用荧光显微镜观察细胞内MPPA的荧光强度，并用微板仪进行定量。用免疫印迹法(WB)检测细胞内蛋白质水平。与亲本细胞相比，MG63/PDT和HOS/PDT对MPPA-PDT的抗性分别是亲本细胞的1.67倍和1.61倍，对顺铂也表现出耐药。流式细胞仪检测证实，MPPA-PDT处理的MG63/PDT和HOS/PDT细胞的凋亡率均显著低于相应的亲本细胞。MG63/PDT和HOS/PDT细胞的凋亡相关蛋白(裂解caspase3和裂解PARP)、细胞内ROS和抗氧化蛋白(HO-1和SOD1)的表达也低于亲代细胞。与MG63和HOS细胞相比，MG63/PDT和HOS/PDT细胞在增殖、光敏剂吸收、集落形成、侵袭、迁移和细胞周期分布等方面均发生了变化。与MG63和HOS细胞相比，两种耐药细胞CD133、Survivin、BclxL、Bcl2、MRP1、MDR1和ABCG2的表达均较高，而Bax的表达较低。本研究成功地建立了两株人骨肉瘤耐药细胞系，为探索耐药相关机制和克服耐药途径提供了有价值的实验材料。
The present study was performed to establish and characterize new human osteosarcoma cell lines resistant to pyropheophorbide-α methyl ester-mediated photodynamic therapy (MPPa-PDT). MPPa-PDT-resistant cells are isolated from the human osteosarcoma MG63 and HOS cell lines and two resistant populations were finally acquired, including MG63/PDT and HOS/PDT. Cell Counting Kit-8 (CCK-8) assay was used to determine the MPPa-PDT, cisplatin (CDDP) resistance and proliferation of MG63, MG63/PDT, HOS and HOS/PDT cells. The intracellular ROS were analyzed using DCFH-DA staining. The colony formation, invasion and migration of parental and resistant cells were compared. FCM was employed to examine the cell cycle distribution, the apoptosis rate and the proportion of CD133+ cells. The fluorescence intensity of intracellular MPPa was observed by fluorescence microscopy and quantified using microplate reader. The protein levels were assessed by western blotting (WB). Compared with two parental cells, MG63/PDT and HOS/PDT were 1.67- and 1.61-fold resistant to MPPa-PDT, respectively, and also exhibited the resistance to CDDP. FCM assays confirmed that both MG63/PDT and HOS/PDT cells treated with MPPa-PDT displayed a significantly lower apoptosis rate in comparison with their corresponding parental cells. The expression of apoptosis-related proteins (i.e. cleaved-caspase 3 and cleaved-PARP), intracellular ROS and the antioxidant proteins (HO-1 and SOD1) in MG63/PDT and HOS/PDT cells was also lower than that in parental cells. Both MG63/PDT and HOS/PDT cells exhibited changes in proliferation, photosensitizer absorption, colony formation, invasion, migration and the cell cycle distribution as compared to MG63 and HOS cells, respectively. Compared to MG63 and HOS cells, both resistant cell lines had a higher expression of CD133, survivin, Bcl-xL, Bcl-2, MRP1, MDR1 and ABCG2, but a lower expression of Bax. The present study successfully established two resistant human osteosarcoma cell lines which are valuable to explore the resistance-related mechanisms and the approaches to overcome resistance.
登录
查看更多内容
CD133表达预测骨肉瘤患者的肺转移和不良预后：临床和实验研究
影响因子：
2.7
作者：
He A;Qi W;Huang Y;Feng T;Chen J;Sun Y;Shen Z;Yao Y
通讯作者：
Yao Y
影响因子：
2.5
作者：
Dean, Michael
通讯作者：
Dean, Michael
DOI：
10.3322/caac.20114
发表时间：
2011-07
期刊：
CA: a cancer journal for clinicians
影响因子：
--
作者：
Agostinis P;Berg K;Cengel KA;Foster TH;Girotti AW;Gollnick SO;Hahn SM;Hamblin MR;Juzeniene A;Kessel D;Korbelik M;Moan J;Mroz P;Nowis D;Piette J;Wilson BC;Golab J
通讯作者：
Golab J
影响因子：
--
作者：
Ishikawa T;Nakagawa H;Hagiya Y;Nonoguchi N;Miyatake S;Kuroiwa T
通讯作者：
Kuroiwa T
影响因子：
9.7
作者：
Casas, Adriana;Di Venosa, Gabriela;Vanzulli, Silvia;Perotti, Christian;Mamome, Leandro;Rodriguez, Lorena;Simian, Marina;Juarranz, Angeles;Pontiggia, Osvaldo;Hasan, Tayyaba;Batlle, Alcira
通讯作者：
Batlle, Alcira