Sirolimus induces apoptosis and reverses multidrug resistance in human osteosarcoma cells in vitro via increasing microRNA-34b expression
Sirolimus induces apoptosis and reverses multidrug resistance in human osteosarcoma cells in vitro via increasing microRNA-34b expression
复制标题
西罗莫司通过增加 microRNA-34b 表达诱导体外人骨肉瘤细胞凋亡并逆转多药耐药性
DOI：
10.1038/aps.2015.153
复制
发表时间：
2016-02
期刊：
影响因子：
--
通讯作者：
haiyan hu
中科院分区：
文献类型：
--
作者：
haiyan hu
Aim:Multi-drug resistance poses a critical bottleneck in chemotherapy. Given the up-regulation of mTOR pathway in many chemoresistant cancers, we examined whether sirolimus (rapamycin), a first generation mTOR inhibitor, might induce human osteosarcoma (OS) cell apoptosis and increase the sensitivity of OS cells to anticancer drugs in vitro.Methods:Human OS cell line MG63/ADM was treated with sirolimus alone or in combination with doxorubicin (ADM), gemcitabine (GEM) or methotrexate (MTX). Cell proliferation and apoptosis were detected using CCK-8 assay and flow cytometry, respectively. MiRNAs in the cells were analyzed with miRNA microarray. The targets of miR-34b were determined based on TargetScan analysis and luciferase reporter assays. The expression of relevant mRNA and proteins was measured using qRT-PCR and Western blotting. MiR-34, PAK1 and ABCB1 levels in 40 tissue samples of OS patients were analyzed using qRT-PCR and in situ hybridization assays.Results:Sirolimus (1–100 nmol/L) dose-dependently suppressed the cell proliferation (IC 50= 23.97 nmol/L) and induced apoptosis. Sirolimus (10 nmol/L) significantly sensitized the cells to anticancer drugs, leading to decreased IC 50 values of ADM, GEM and MTX (from 25.48, 621.41 and 21.72 μmol/L to 4.93, 73.92 and 6.77 μmol/L, respectively). Treatment of with sirolimus increased miR-34b levels by a factor of 7.5 in the cells. Upregulation of miR-34b also induced apoptosis and increased the sensitivity of the cells to the anticancer drugs, whereas transfection with miR-34b-AMO, an inhibitor of miR-34b, reversed the anti-proliferation effect of sirolimus. Two key regulators of cell cycle, apoptosis and multiple drug resistance, PAK1 and ABCB1, were demonstrated to be the direct targets of miR-34b. In 40 tissue samples of OS patients, significantly higher miR-34 ISH score and lower PAK5 and ABCB1 scores were detected in the chemo-sensitive group.Conclusion:Sirolimus increases the sensitivity of human OS cells to anticancer drugs in vitro by up-regulating miR-34b interacting with PAK1 and ABCB1. A low miR-34 level is an indicator of poor prognosis in OS patients.
登录
查看更多内容
影响因子：
3.2
作者：
Bao B;Li Y;Ahmad A;Azmi AS;Bao G;Ali S;Banerjee S;Kong D;Sarkar FH
通讯作者：
Sarkar FH
影响因子：
9.7
作者：
W. Utomo;Vilvapathy Narayanan;K. Biermann;C. V. van Eijck;M. Bruno;M. Peppelenbosch;H. Braat
通讯作者：
W. Utomo;Vilvapathy Narayanan;K. Biermann;C. V. van Eijck;M. Bruno;M. Peppelenbosch;H. Braat
DOI：
10.1186/bcr3640
发表时间：
2014-04-07
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Zhang H;Cohen AL;Krishnakumar S;Wapnir IL;Veeriah S;Deng G;Coram MA;Piskun CM;Longacre TA;Herrler M;Frimannsson DO;Telli ML;Dirbas FM;Matin AC;Dairkee SH;Larijani B;Glinsky GV;Bild AH;Jeffrey SS
通讯作者：
Jeffrey SS
MTOR 抑制通过自噬介导的 CREB1 抑制减轻 DNA 损伤和细胞凋亡
影响因子：
13.3
作者：
Wang, Ying;Hu, Zhongdong;Zhang, Hongbing
通讯作者：
Zhang, Hongbing
影响因子：
9.7
作者：
Ye, Zhimin;Fang, Jun;Huang, Pintong
通讯作者：
Huang, Pintong