Stable SET knockdown in head and neck squamous cell carcinoma promotes cell invasion and the mesenchymal-like phenotype in vitro, as well as necrosis, cisplatin sensitivity and lymph node metastasis in xenograft tumor models.
Stable SET knockdown in head and neck squamous cell carcinoma promotes cell invasion and the mesenchymal-like phenotype in vitro, as well as necrosis, cisplatin sensitivity and lymph node metastasis in xenograft tumor models.
复制标题
DOI：
10.1186/1476-4598-13-32
复制
发表时间：
2014-02-20
期刊：
影响因子：
37.3
通讯作者：
Leopoldino AM
中科院分区：
文献类型：
--
作者：
Sobral LM;Sousa LO;Coletta RD;Cabral H;Greene LJ;Tajara EH;Gutkind JS;Curti C;Leopoldino AM
SET/I2 PP 2A是一种在头颈部鳞状细胞癌（HNSCC）中上调的多功能蛋白。SET在HNSCC致瘤性中的作用尚不清楚。在HN 12、HN 13和Cal 27三种HNSCC细胞系中建立了稳定的通过shRNA的SET敲低（shSET）。蛋白质表达和磷酸化的蛋白质水平通过Western印迹和免疫荧光法测定，细胞迁移和侵袭通过功能分析测定，PP 2A活性使用丝氨酸/苏氨酸磷酸酶测定。使用实时PCR阵列定量与细胞运动相关的84个基因。金属蛋白酶（MMP）活性通过酶谱法和荧光测定法进行评估。在Balb/c裸鼠中建立HN 12 shSET异种移植瘤（侧腹和舌模型）;通过肉眼、免疫组织化学和组织学分析证明异种移植瘤特征和顺铂敏感性，以及通过组织学证明淋巴结转移。HN 1/2shSET细胞与对照组相比，ERK 1/2和p53磷酸化水平降低。ShSET抑制HN 12细胞增殖并增加HN 12和Cal 27细胞的亚G1群体。PP 2A活性增加也与shSET相关。PCR芯片显示波形蛋白、基质金属蛋白酶9（MMP 9）和非肌球蛋白重链IIB在HN 12细胞中表达上调。减少E-钙粘蛋白和泛细胞角蛋白，以及增加波形蛋白，也被证明是SET敲低的结果。这些变化伴随着MMP-9和MMP-2活性的增加、迁移和侵袭。HN 12 shSET皮下异种移植肿瘤表现出分化差的表型、细胞增殖降低和顺铂敏感性。使用HN 12 shSET细胞的原位异种移植肿瘤模型显示出增加的转移潜力。SET积累在HNSCC中起重要作用。作为癌基因，SET在体内促进细胞增殖、存活和对顺铂引起的细胞死亡的抵抗。作为转移抑制因子，SET调节侵袭、上皮间质转化和转移。
SET/I2PP2A is a multifunctional protein that is up-regulated in head and neck squamous cell carcinoma (HNSCC). The action of SET in HNSCC tumorigenicity is unknown. Stable SET knockdown by shRNA (shSET) was established in three HNSCC cell lines: HN12, HN13, and Cal27. Protein expression and phosphorylated protein levels were determined by Western blotting and immunofluorescence, cell migration and invasion were measured by functional analysis, and PP2A activity was determined using a serine/threonine phosphatase assay. A real-time PCR array was used to quantify 84 genes associated with cell motility. Metalloproteinase (MMP) activity was assessed by zymographic and fluorometric assays. HN12shSET xenograft tumors (flank and tongue models) were established in Balb/c nude mice; the xenograft characteristics and cisplatin sensitivity were demonstrated by macroscopic, immunohistochemical, and histological analyses, as well as lymph node metastasis by histology. The HN12shSET cells displayed reduced ERK1/2 and p53 phosphorylation compared with control. ShSET reduced HN12 cell proliferation and increased the sub-G1 population of HN12 and Cal27 cells. Increased PP2A activity was also associated with shSET. The PCR array indicated up-regulation of three mRNAs in HN12 cells: vimentin, matrix metalloproteinase-9 (MMP9) and non-muscle myosin heavy chain IIB. Reduced E-cadherin and pan-cytokeratin, as well as increased vimentin, were also demonstrated as the result of SET knockdown. These changes were accompanied by an increase in MMP-9 and MMP-2 activities, migration and invasion. The HN12shSET subcutaneous xenograft tumors presented a poorly differentiated phenotype, reduced cell proliferation, and cisplatin sensitivity. An orthotopic xenograft tumor model using the HN12shSET cells displayed increased metastatic potential. SET accumulation has important actions in HNSCC. As an oncogene, SET promotes cell proliferation, survival, and resistance to cell death by cisplatin in vivo. As a metastasis suppressor, SET regulates invasion, the epithelial mesenchymal transition, and metastasis.
登录
查看更多内容
DOI：
10.1002/hed.10274
发表时间：
2003-08-01
影响因子：
2.9
作者：
Bradford, CR;Zhu, SB;Carey, TE
通讯作者：
Carey, TE
影响因子：
3.5
作者：
Lam, B. Daniel;Anthony, Eloise C.;Hordijk, Peter L.
通讯作者：
Hordijk, Peter L.
DOI：
10.1083/jcb.201009059
发表时间：
2011-01-24
期刊：
The Journal of cell biology
影响因子：
--
作者：
Muller PA;Vousden KH;Norman JC
通讯作者：
Norman JC
影响因子：
4.8
作者：
Kuo, Yi-Chun;Huang, Kai-Yun;Chiang, Chi-Wu
通讯作者：
Chiang, Chi-Wu
影响因子：
56.9
作者：
Ghosh, M;Song, XY;Condeelis, JS
通讯作者：
Condeelis, JS