Establishment of stable multiple myeloma cell line with overexpressed PDCD5 and its proapoptosis mechanism.
Establishment of stable multiple myeloma cell line with overexpressed PDCD5 and its proapoptosis mechanism.
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PDCD5过表达稳定多发性骨髓瘤细胞系的建立及其促凋亡机制
DOI：
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发表时间：
2015-09
期刊：
影响因子：
--
通讯作者：
Liu Jing
中科院分区：
文献类型：
--
作者：
Feng Wenchang;Fu Yunfeng;Zhang Yanan;Lv Ben;Li Xin;Zhang Fan;Gui Rong;Liu Jing
目的
建立了表现出稳定的多西环素（DOX）诱导的PDCD 5表达的转染的多发性骨髓瘤细胞系。分析PDCD 5过表达对地塞米松（DXM）诱导的细胞凋亡的影响沿着探讨其机制。
方法
(1)用慢病毒载体将PDCD 5基因转染到多发性骨髓瘤细胞中。通过应用嘌呤霉素进行筛选，并通过DOX诱导PDCD 5表达。（2）Annexin-APC/PI双染色流式细胞仪检测稳定转染组、空白组和空载体组细胞凋亡率;（3）采用实时荧光定量PCR和Western Blot方法检测Survivin、casepase-3和Bcl-2基因和蛋白的表达水平。
结果
与空白组和空载体组相比，稳定转染的多发性骨髓瘤细胞PDCD 5表达明显增加。转染组细胞对DXM敏感，凋亡细胞比例明显高于空白组和空载体组（P<0.05）。Survivin和Bcl-2在U266/PDCD 5细胞和联合DXM组中表达较单药组明显下调。然而，caspase-3显著上调。
结论
建立了内源性PDCD 5基因转染的多发性骨髓瘤细胞系。内源性PDCD 5过表达促进DXM诱导的细胞凋亡。PDCD 5基因的促凋亡作用具有激活case 3、下调survivin和Bcl-2的作用，进一步促进多发性骨髓瘤细胞凋亡。
OBJECTIVE
The transfected multiple myeloma cell line showing a stable doxycycline (DOX)-induced expression of PDCD5 was established. PDCD5 overexpression in the transfected cell line was analyzed for its effect on the dexamethasone (DXM)-induced apoptosis along with a discussion on the mechanism.
METHODS
(1) Lentiviral plasmid was used for the transfection of PDCD5 gene into the multiple myeloma cells. The screening was done by applying puromycin, and PDCD5 expression was induced by DOX. Real-time fluorescence quantitative PCR and Western Blot were performed to detect the expression levels of the target gene in the stable transfection group and the empty vector group; (2) The cell apoptosis rates of stable transfection group, blank group and empty vector group were measured by Annexin-APC/PI double staining flow cytometry; (3) Real-time fluorescence quantitative PCR and Western Blot were carried out to detect the expression levels of survivin, casepase-3 and Bcl-2 genes and proteins.
RESULTS
PDCD5 expression was significantly increased in the stably tranfected multiple myeloma cells compared with blank group and empty vector group. The cells in the transfection group were more sensitive to DXM, and the proportion of apoptotic cells was obviously higher than that of the blank group and the empty vector group (P<0.05). Survivin and Bcl-2 were considerably downregulated in U266/PDCD5 cells and combined DXM group than in the single agent group. However, caspase-3 was significantly upregulated.
CONCLUSION
Multiple myeloma cell line transfected with endogenous PDCD5 gene was established. The endogenous PDCD5 overexpression accelerated the cell apoptosis under DXM induction. The proapoptotic action of PDCD5 gene had the effect of activating casepase-3 and downregulating survivin and Bcl-2, which further promoted the apoptosis of multiple myeloma cells.
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DOI：
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发表时间：
2004-08
影响因子：
--
作者：
Zhaohui Liu;Dai Zhang;Ke-min Li;Q. Liao
通讯作者：
Zhaohui Liu;Dai Zhang;Ke-min Li;Q. Liao
重组人PDCD5在体外和体内使软骨肉瘤对顺铂化疗敏感
影响因子：
7.2
作者：
Chen, Changbao;Zhou, Hua;Chen, Yingyu
通讯作者：
Chen, Yingyu
影响因子：
3.7
作者：
Waligórska-Stachura J;Andrusiewicz M;Sawicka-Gutaj N;Biczysko M;Jankowska A;Kubiczak M;Czarnywojtek A;Wrotkowska E;Ruchała M
通讯作者：
Ruchała M
影响因子：
4.2
作者：
H. Ben‐Hur;Eitan Mordechay;R. Halperin;P. Gurevich;J. Zandbank;Meherdad Herper;I. Zusman
通讯作者：
H. Ben‐Hur;Eitan Mordechay;R. Halperin;P. Gurevich;J. Zandbank;Meherdad Herper;I. Zusman
DOI：
10.1073/pnas.93.21.11382
发表时间：
1996-10-15
影响因子：
11.1
作者：
Naldini, L;Blomer, U;Verma, IM
通讯作者：
Verma, IM