Restoration of WNT4 inhibits cell growth in leukemia-derived cell lines.
Restoration of WNT4 inhibits cell growth in leukemia-derived cell lines.
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DOI：
10.1186/1471-2407-13-557
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发表时间：
2013-11-25
期刊：
影响因子：
3.8
通讯作者：
Aguilar-Lemarroy A
中科院分区：
文献类型：
--
作者：
García-Castro B;Alvarez-Zavala M;Riveros-Magaña AR;Ortíz-Lazareno PC;Ratkovich-González S;Hernández-Flores G;Bravo-Cuellar A;Jave-Suarez LF;Aguilar-Lemarroy A
在癌症的发展过程中，WNT信号通路发生了显著变化。脊椎动物有两类Wnt信号通路：Wnt/β-catenin信号通路和Wnt/Ca~(2+)和Wnt/平面细胞极性[PCP]信号通路。Wnt4影响造血祖细胞的增殖和存活；然而，Wnt4在癌症发展中的作用及其对肿瘤发生的影响还知之甚少。本研究的目的有两个方面：第一，检测WNT4在成熟外周血细胞和各种白血病来源细胞中的表达，包括血液肿瘤细胞系和白血病患者的细胞系；第二，确定该配体对原始细胞来源的BJAB、Jurkat、CEM、K562和HL60增殖和凋亡的影响。我们用定量逆转录聚合酶链式反应(qRT-PCR)检测了正常人外周血单个核细胞(PBMC)和T、B淋巴细胞以及5个白血病来源的细胞系和白血病患者来源的原始细胞中WNT4的表达。为了分析WNT4对细胞增殖的影响，将PBMC和细胞系暴露于商业上可获得的WNT4重组人蛋白中。此外，使用可诱导的慢病毒表达系统在BJAB细胞中恢复了WNT4的表达。通过向细胞培养中加入WST-1和计数细胞来检测细胞的存活和增殖；此外，在没有或存在WNT4的情况下，分析细胞周期的进展和凋亡率。采用定量RT-PCR方法检测WNT途径靶基因的表达。在白血病来源的细胞系和白血病患者来源的原始细胞中，WNT4的表达严重减少。细胞株暴露于WNT4重组蛋白显著抑制细胞增殖；诱导BJAB细胞表达WNT4证实了这一观察结果。有趣的是，Wnt4在BJAB细胞中的表达恢复增加了细胞在G1期的积聚，而不会诱导规范的Wnt/β-catenin靶基因的激活。我们的研究结果表明，Wnt4配体通过非依赖于FZD6的方式将细胞阻滞在G1期，可能是通过拮抗经典的Wnt/β-catenin信号通路，从而在调节白血病细胞的生长中发挥作用。
WNT signaling pathways are significantly altered during cancer development. Vertebrates possess two classes of WNT signaling pathways: the “canonical” WNT/β-catenin signaling pathway, and the “non-canonical” pathways including WNT/Ca2+ and WNT/Planar cell polarity [PCP] signaling. WNT4 influences hematopoietic progenitor cell expansion and survival; however, WNT4 function in cancer development and the resulting implications for oncogenesis are poorly understood. The aim of this study was twofold: first, to determine the expression of WNT4 in mature peripheral blood cells and diverse leukemia-derived cells including cell lines from hematopoietic neoplasms and cells from patients with leukemia; second, to identify the effect of this ligand on the proliferation and apoptosis of the blast-derived cell lines BJAB, Jurkat, CEM, K562, and HL60. We determined WNT4 expression by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) in peripheral blood mononuclear cells (PBMCs) and T- and B-lymphocytes from healthy individuals, as well as from five leukemia-derived cell lines and blasts derived from patients with leukemia. To analyze the effect of WNT4 on cell proliferation, PBMCs and cell lines were exposed to a commercially available WNT4 recombinant human protein. Furthermore, WNT4 expression was restored in BJAB cells using an inducible lentiviral expression system. Cell viability and proliferation were measured by the addition of WST-1 to cell cultures and counting cells; in addition, the progression of the cell cycle and the amount of apoptosis were analyzed in the absence or presence of WNT4. Finally, the expression of WNT-pathway target genes was measured by qRT-PCR. WNT4 expression was severely reduced in leukemia-derived cell lines and blasts derived from patients with leukemia. The exposure of cell lines to WNT4 recombinant protein significantly inhibited cell proliferation; inducing WNT4 expression in BJAB cells corroborated this observation. Interestingly, restoration of WNT4 expression in BJAB cells increased the accumulation of cells in G1 phase, and did not induce activation of canonical WNT/β-catenin target genes. Our findings suggest that the WNT4 ligand plays a role in regulating the cell growth of leukemia-derived cells by arresting cells in the G1 cell cycle phase in an FZD6-independent manner, possibly through antagonizing the canonical WNT/β-catenin signaling pathway.
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影响因子：
2.7
作者：
Sercan, Zeynep;Pehlivan, Melek;Sercan, Hakki Ogun
通讯作者：
Sercan, Hakki Ogun
影响因子：
4.7
作者：
Spinsanti, Paola;De Vita, Teresa;Nicoletti, Ferdinando
通讯作者：
Nicoletti, Ferdinando
影响因子：
6.5
作者：
Bonifas, JM;Pennypacker, S;Epstein, EH
通讯作者：
Epstein, EH
影响因子：
20.3
作者：
Heinonen, Krista M.;Vanegas, Juan Ruiz;Perreault, Claude
通讯作者：
Perreault, Claude
影响因子：
2.7
作者：
Bernard, Pascal;Fleming, Alice;Vilain, Eric
通讯作者：
Vilain, Eric