浏览全部资源
扫码关注微信
FABD结构域缺失削弱293T细胞BCR/ABL癌蛋白抗凋亡能力
Depletion of F-actin binding domain reduces the ability of BCR/ABL to resist apoptosis in 293T cells
- 第三军医大学学报 2018年40卷第17期 页码：1548-1553
- 作者机构：
- 作者简介：
- 基金信息：国家自然科学基金面上项目(81572060);重庆市研究生创新项目(CYS16136)
DOI：10.16016/j.1000-5404.201802024
中图分类号：收稿：2018-02-04，
纸质出版：2018-09-15
- 稿件说明：
移动端阅览
周芳竹, 黄峥兰, 骆贞红, 等. FABD结构域缺失削弱293T细胞BCR/ABL癌蛋白抗凋亡能力[J]. 第三军医大学学报, 2018,40(17):1548-1553.
Fangzhu ZHOU, Zhenglan HUANG, Zhenhong LUO, et al. Depletion of F-actin binding domain reduces the ability of BCR/ABL to resist apoptosis in 293T cells[J]. Journal of Third Military Medical University, 2018, 40(17): 1548-1553.
周芳竹, 黄峥兰, 骆贞红, 等. FABD结构域缺失削弱293T细胞BCR/ABL癌蛋白抗凋亡能力[J]. 第三军医大学学报, 2018,40(17):1548-1553. DOI： 10.16016/j.1000-5404.201802024.
Fangzhu ZHOU, Zhenglan HUANG, Zhenhong LUO, et al. Depletion of F-actin binding domain reduces the ability of BCR/ABL to resist apoptosis in 293T cells[J]. Journal of Third Military Medical University, 2018, 40(17): 1548-1553. DOI： 10.16016/j.1000-5404.201802024.
摘要
Abstract
Objective
2
To investigate the effects of F-actin binding domain (FABD) on BCR/ABL function in 293T cells by transfecting wild-type pAd-Bcr/Abl and mutant pAd-Bcr/Abl -ΔFABD.
Methods
2
Firstly
293T cells were transfected wild-type pAd-Bcr/Abl and mutant pAd-Bcr/Abl-ΔFABD plasmid
respectively. Then the transfected cells and blank control cells were cultivated for 24
48
or 72 h. Flow cytometry was used to detect the fluorescence intensity to screen optimal transfection conditions. Western blotting was employed to detect the expression of related proteins. Secondly
the effects of FABD on proliferation and apoptosis of the cells promoted by BCR/ABL oncoprotein were detected by WST-1 assay and flow cytometry
respectively. DAPI staining was adopted to observe the changes of nuclear morphology. The protein expression of p-BCR/ABL
p-AKT
p-ERK
and apoptosis related protein Caspase-3
and PARP activity was determined by Western blotting.
Results
2
The results of flow cytometry showed that the efficiency of plas
mid transfection was increased in a time-dependent manner
and 48 h was identified as the optimal transfection time. Western blotting results confirmed the pAd-Bcr/Abl and mutant pAd-Bcr/Abl-ΔFABD plasmids were constructed successfully. After the transfection of pAd-Bcr/Abl and mutant pAd-Bcr/Abl-ΔFABD plasmids into 293T cells
WST-1 assay and flow cytometry that FABD depletion significant inhibited cell proliferation and promoted apoptosis
with the percentage of apoptotic cells accounting for 29.0% in 48 h after treatment (
P
<
0.05). The depletion also down-regulated the expression of p-AKT and p-ERK and activated Caspase-3 and PARP in 48 h after transfection.
Conclusion
2
FABD depletion reduces the cell apoptosis promoted by BCR/ABL oncoprotein
which might be though down-regulating AKT and ERK signaling pathways and activating mitochondrial associated apoptotic pathway.
关键词
Keywords
references
KANG Z J LIU Y F XU L Z The Philadelphia chromosome in leukemogenesis Chin J Cancer 2016 35 48 10.1186/s40880-016-0108-0 KANG Z J, LIU Y F, XU L Z, et al . The Philadelphia chromosome in leukemogenesis[J ] . Chin J Cancer, 2016, 35: 48. DOI:10.1186/s40880-016-0108-0.
APPERLEY J F Chronic myeloid leukaemia Lancet 2015 385 9976 1447 1459 10.1016/S0140-6736(13)62120-0 APPERLEY J F. Chronic myeloid leukaemia[J]. Lancet, 2015, 385(9976):1447-1459. DOI:10.1016/S0140-6736(13)62120-0.
SOVERINI S MANCINI M BAVARO L Chronic myeloid leukemia: the paradigm of targeting oncogenic tyrosine kinase signaling and counteracting resistance for successful cancer therapy Mol Cancer 2018 17 1 49 10.1186/s12943-018-0780-6 SOVERINI S, MANCINI M, BAVARO L, et al . Chronic myeloid leukemia: the paradigm of targeting oncogenic tyrosine kinase signaling and counteracting resistance for successful cancer therapy[J ] . Mol Cancer, 2018, 17(1): 49. DOI:10.1186/s12943-018-0780-6.
SAMASSEKOU O NTWARI A HÉBERT J Individual telomere lengths in chronic myeloid leukemia Neoplasia 2009 11 11 1146 1154 10.1182/blood-2008-03-144790 SAMASSEKOU O, NTWARI A, HÉBERT J, et al . Individual telomere lengths in chronic myeloid leukemia[J ] . Neoplasia, 2009, 11(11):1146-1154. DOI:10.1182/blood-2008-03-144790.
MA L XU Z WANG J Matrine inhibits BCR/ABL mediated ERK/MAPK pathway in human leukemia cells Oncotarget 2017 8 65 108880 108889 10.18632/oncotarget.22353 MA L, XU Z, WANG J, et al . Matrine inhibits BCR/ABL mediated ERK/MAPK pathway in human leukemia cells[J ] . Oncotarget, 2017, 8(65):108880-108889. DOI:10.18632/oncotarget.22353.
HUANG R LIU H CHEN Y EPS8 regulates proliferation, apoptosis and chemosensitivity in BCR-ABL positive cells via the BCR-ABL/PI3K/AKT/mTOR pathway Oncol Rep 2018 39 1 119 128 10.3892/or.2017.6102 HUANG R, LIU H, CHEN Y, et al . EPS8 regulates proliferation, apoptosis and chemosensitivity in BCR-ABL positive cells via the BCR-ABL/PI3K/AKT/mTOR pathway[J ] . Oncol Rep, 2018, 39(1): 119-128. DOI:10.3892/or.2017.6102.
PADULA W V LARSON R A DUSETZINA S B Cost-effectiveness of tyrosine kinase inhibitor treatment strategies for chronic myeloid leukemia in chronic phase after generic entry of Imatinib in the United States J Natl Cancer Inst 2016 108 7 djw003 10.1093/jnci/djw003 PADULA W V, LARSON R A, DUSETZINA S B, et al . Cost-effectiveness of tyrosine kinase inhibitor treatment strategies for chronic myeloid leukemia in chronic phase after generic entry of Imatinib in the United States[J ] . J Natl Cancer Inst, 2016, 108(7): djw003. DOI: 10.1093/jnci/djw003.
STAGNO F STELLA S SPITALERI A Imatinib mesylate in chronic myeloid leukemia: frontline treatment and long-term outcomes Expert Rev Anticancer Ther 2016 16 3 273 278 10.1586/14737140.2016.1151356 STAGNO F, STELLA S, SPITALERI A, et al . Imatinib mesylate in chronic myeloid leukemia: frontline treatment and long-term outcomes[J ] . Expert Rev Anticancer Ther, 2016, 16(3): 273-278. DOI:10.1586/14737140.2016.1151356.
PATKAR N GHODKE K JOSHI S Characteristics of BCR-ABL kinase domain mutations in chronic myeloid leukemia from India: not just missense mutations but insertions and deletions are also associated with TKI resistance Leuk Lymphoma 2016 57 11 2653 2660 10.3109/10428194.2016.1157868 PATKAR N, GHODKE K, JOSHI S, et al . Characteristics of BCR-ABL kinase domain mutations in chronic myeloid leukemia from India: not just missense mutations but insertions and deletions are also associated with TKI resistance[J ] . Leuk Lymphoma, 2016, 57(11): 2653-2660. DOI:10.3109/10428194.2016.1157868.
PREYER M VIGNERI P WANG J Y Interplay between kinase domain autophosphorylation and F-actin binding domain in regulating imatinib sensitivity and nuclear import of BCR-ABL PLoS ONE 2011 6 2 e17020 10.1371/journal.pone.0017020 PREYER M, VIGNERI P, WANG J Y. Interplay between kinase domain autophosphorylation and F-actin binding domain in regulating imatinib sensitivity and nuclear import of BCR-ABL[J]. PLoS ONE, 2011, 6(2): e17020. DOI:10.1371/journal.pone.0017020.
HANTSCHEL O WIESNER S GüTTLER T Structural basis for the cytoskeletal association of Bcr-Abl/c-Abl Mol Cell 2005 19 4 461 473 10.1016/j.molcel.2005.06.030 HANTSCHEL O, WIESNER S, GüTTLER T, et al . Structural basis for the cytoskeletal association of Bcr-Abl/c-Abl[J ] . Mol Cell, 2005, 19(4): 461-473. DOI:10.1016/j.molcel.2005.06.030.
LI Y CLOUGH N SUN X Bcr-Abl induces abnormal cytoskeleton remodeling, beta1 integrin clustering and increased cell adhesion to fibronectin through the Abl interactor 1 pathway J Cell Sci 2007 120 Pt 8 1436 1446 10.1242/jcs.03430 LI Y, CLOUGH N, SUN X, et al . Bcr-Abl induces abnormal cytoskeleton remodeling, beta1 integrin clustering and increased cell adhesion to fibronectin through the Abl interactor 1 pathway[J ] . J Cell Sci, 2007, 120(Pt 8): 1436-1446. DOI:10.1242/jcs.03430.
IZDEBSKA M ZIELIŃSKA W GRZANKA D The Role of actin dynamics and actin-binding proteins expression in epithelial-to-mesenchymal transition and its association with cancer progression and evaluation of possible therapeutic targets Biomed Res Int 2018 2018 4578373 10.1155/2018/4578373 IZDEBSKA M, ZIELIŃSKA W, GRZANKA D, et al . The Role of actin dynamics and actin-binding proteins expression in epithelial-to-mesenchymal transition and its association with cancer progression and evaluation of possible therapeutic targets[J ] . Biomed Res Int, 2018, 2018: 4578373. DOI:10.1155/2018/4578373.
KUŽELOVÁ K PLUSKALOVÁ M GREBEŇOVÁ D Changes in cell adhesivity and cytoskeleton-related proteins during imatinib-induced apoptosis of leukemic JURL-MK1 cells J Cell Biochem 2010 111 6 1413 1425 10.1002/jcb.22868 KUŽELOVÁ K, PLUSKALOVÁ M, GREBEŇOVÁ D, et al . Changes in cell adhesivity and cytoskeleton-related proteins during imatinib-induced apoptosis of leukemic JURL-MK1 cells[J ] . J Cell Biochem, 2010, 111(6): 1413-1425. DOI:10.1002/jcb.22868.
王 腾 黄 峥兰 高 淼 探讨FABD结构域对Bcr/Abl蛋白定位及功能的影响 重庆医科大学学报 2017 42 11 1477 1483 10.13406/j.cnki.cyxb.001405 王腾, 黄峥兰, 高淼, 等.探讨FABD结构域对Bcr/Abl蛋白定位及功能的影响[J].重庆医科大学学报, 2017, 42(11):1477-1483.DOI:10.13406/j.cnki.cyxb.001405.
WANG T HUANG Z L GAO M Effect of FABD on Bcr/Abl cytoplasm localization and function in 293T cells J Chongqing Med Univ 2017 42 11 1477 1483 10.13406/j.cnki.cyxb.001405 WANG T, HUANG Z L, GAO M, et al .Effect of FABD on Bcr/Abl cytoplasm localization and function in 293T cells[J ] . J Chongqing Med Univ, 2017, 42(11):1477-1483.DOI:10.13406/j.cnki.cyxb.001405.
MITRA A RADHA V F-actin-binding domain of c-Abl regulates localized phosphorylation of C3G: role of C3G in c-Abl-mediated cell death Oncogene 2010 29 32 4528 4542 10.1038/onc.2010.113 MITRA A, RADHA V. F-actin-binding domain of c-Abl regulates localized phosphorylation of C3G: role of C3G in c-Abl-mediated cell death[J]. Oncogene, 2010, 29(32): 4528-4542. DOI:10.1038/onc.2010.113.
0
浏览量
0
下载量
0
CSCD
- Meta-XML下载
- BibTeX下载
- Endnote下载
- RefMan 下载
- NoteExpress下载
- NoteFirst下载
关联资源
相关文章
相关作者
相关机构
- 技术支持由北京北大方正电子有限公司提供 京ICP备09064830号-19
京公网安备11010802024621 - 本系统建议在Chrome、 IE9+ 以上版本浏览器阅读本站内容，360浏览器请切换至极速模式
- Cookies帮助我们提供服务并提供个性化体验。使用本网站，即表示您同意我们使用Cookies
