南方医科大学学报

南方医科大学学报 ›› 2017, Vol. 37 ›› Issue (12): 1654-.

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过表达脾酪氨酸激酶通过调控Fra-1抑制结直肠癌细胞的增殖和促进其凋亡

席作武，梁伟涛   

  • 出版日期:2017-12-20 发布日期:2017-12-20

Spleen tyrosine kinase inhibits proliferation and promotes apoptosis of colorectal cancer cells in vitro via regulating Fra-1

  • Online:2017-12-20 Published:2017-12-20

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摘要： 目的探讨过表达脾酪氨酸激酶（SYK）对结直肠癌细胞增殖和凋亡的影响及可能的相关机制。方法利用pcDNA.3.1质 粒构建重组质粒pcDNA.3.1-SYK，转染结直肠癌细胞，过表达SYK，分组情况如下。（1）pcDNA.3.1-SYK（HCT116）：转染 pcDNA.3.1-SYK 到HCT116；（2）pcDNA.3.1（HCT116）：转染pcDNA.3.1 空载体到HCT116 中；（3）Normal（HCT116）：正常 HCT116 细胞。（1）pcDNA.3.1-SYK（Sw480）：转染pcDNA.3.1-SYK到Sw480；（2）pcDNA.3.1（Sw480）：转染pcDNA.3.1 空载体 到Sw480中；（3）Normal（Sw480）：正常Sw480细胞。应用qRT-PCR法检测结直肠癌和癌旁组织中SYK和Fra-1的mRNA表达 量；Western blot法检测SYK和Fra-1的蛋白表达量；MTT法检测细胞生长活力；BrdU方法检测细胞增殖活性；试剂盒方法检测 Caspase-3的活性；Annexin-V FITC/PI法检测细胞凋亡情况。结果SYK在结直肠癌组织和结直肠癌细胞系中的表达量均降低 （P<0.01）；pcDNA.3.1-SYK转染结直肠癌细胞系，SYK的mRNA（P<0.01）和蛋白表达量显著升高（P<0.01），显示SYK过表达成 功；SYK过表达后结直肠癌细胞生长活力和增值活性显著降低（P<0.01），细胞凋亡增加（P<0.01）；另外，SYK过表达后，Fra-1的 表达量显著被抑制（P<0.01）。结论过表达SYK对结直肠癌细胞的增殖有抑制作用，并且促进结直肠癌细胞的凋亡，其机制有 可能与SYK对Fra-1的调控有关，为结直肠癌的预防和治疗提供参考价值和理论基础。

Abstract: Objective To investigate the effects of spleen tyrosine kinase (SYK) overexpression on proliferation and apoptosis of colorectal cancer cells and explore the possible mechanism. Methods The mRNA expressions of SYK and Fra-1 in 10 clinical specimens of colorectal cancer and 10 adjacent tissues were measured with qRT-PCR, and their protein expressions were detected with Western blotting. The recombinant plasmid pcDNA.3.1-SYK was constructed and transfected into colorectal cancer cells to induce SYK overexpression, and the cell viability and proliferation were assessed using by MTT assay and BrdU assay, respectively; caspase-3 activity in the cells was evaluated with a commercial kit and the cell apoptosis was analyzed with Annexin-V FITC/PI assay. Results The expressions of SYK were significantly decreased in colorectal cancer tissues and colorectal cancer cell lines. Transfection of pcDNA.3.1-SYK into the colorectal cancer cells induced obviously upregulated mRNA and protein expressions of SYK, which caused a significant suppression of the cell viability and proliferation and enhancement of the cell apoptosis along with a significant inhibition of Fra-1 expression. Conclusions SYK overexpression inhibits the proliferation and promotes apoptosis of colorectal cancer cells, and these effects are possibly mediated by the regulation of Fra-1 expression by SYK.