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
发布时间：2019-05-29 01:20
【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q RT-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;Brd U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(P0.01);pcDNA.3.1-SYK转染结直肠癌细胞系,SYK的mRNA(P0.01)和蛋白表达量显著升高(P0.01),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(P0.01),细胞凋亡增加(P0.01);另外,SYK过表达后,Fra-1的表达量显著被抑制(P0.01)。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 effect of overexpression of spleen tyrosine kinase (SYK) on proliferation and apoptosis of colorectal cancer cells and its possible mechanism. Methods the recombinant plasmid pcDNA.3.1-SYK, was constructed by pcDNA.3.1 plasmid and transfected into colorectal cancer cells. The overexpression of SYK, was divided into the following groups. (1) pcDNA.3.1-SYK (HCT116): pcDNA.3.1-SYK into HCT116; (2) pcDNA.3.1 (HCT116): transfer pcDNA.3.1 empty vector into HCT116; (3) Normal (HCT116): normal HCT116 cells. (1) pcDNA.3.1-SYK (Sw480): pcDNA.3.1-SYK to Sw480; (2) pcDNA.3.1 (Sw480): pcDNA.3.1 empty vector was transferred into Sw480, (3) Normal (Sw480): normal Sw480 cells. The mRNA expression of SYK and Fra-1 in colorectal cancer and paracancerous tissues was detected by Q RT-PCR assay. The protein expression of SYK and Fra-1 was detected by; Western blot assay, and the cell proliferation activity was detected by MTT assay.; Brd U assay was used to detect the proliferation activity of colorectal cancer and paracancerous tissues. The activity of Caspase-3 was detected by kit method and apoptosis was detected by Annexin-V FITC/PI assay. Results the expression of SYK in colorectal cancer tissue and colorectal cancer cell line decreased (P 0.01). The mRNA (P01) and protein expression of SYK were significantly increased (P01) in colorectal cancer cell line pcDNA.3.1-SYK, which showed that the overexpression of SYK was successful. After overexpression of SYK, the growth activity and value-added activity of colorectal cancer cells decreased significantly (P01), and apoptosis increased (P01). In addition, after overexpression of SYK, the expression of Fra-1 was significantly inhibited (P01). Conclusion overexpression of SYK can inhibit the proliferation of colorectal cancer cells and promote the apoptosis of colorectal cancer cells. The mechanism may be related to the regulation of Fra-1 by SYK, which provides a reference value and theoretical basis for the prevention and treatment of colorectal cancer.
【作者单位】： 河南省中医院肛肠科;河南中医药大学中医外科;
【基金】：河南省中医药科学研究专项课题项目(2015ZY02049)
【分类号】：R735.3
[Abstract]:Objective to investigate the effect of overexpression of spleen tyrosine kinase (SYK) on proliferation and apoptosis of colorectal cancer cells and its possible mechanism. Methods the recombinant plasmid pcDNA.3.1-SYK, was constructed by pcDNA.3.1 plasmid and transfected into colorectal cancer cells. The overexpression of SYK, was divided into the following groups. (1) pcDNA.3.1-SYK (HCT116): pcDNA.3.1-SYK into HCT116; (2) pcDNA.3.1 (HCT116): transfer pcDNA.3.1 empty vector into HCT116; (3) Normal (HCT116): normal HCT116 cells. (1) pcDNA.3.1-SYK (Sw480): pcDNA.3.1-SYK to Sw480; (2) pcDNA.3.1 (Sw480): pcDNA.3.1 empty vector was transferred into Sw480, (3) Normal (Sw480): normal Sw480 cells. The mRNA expression of SYK and Fra-1 in colorectal cancer and paracancerous tissues was detected by Q RT-PCR assay. The protein expression of SYK and Fra-1 was detected by; Western blot assay, and the cell proliferation activity was detected by MTT assay.; Brd U assay was used to detect the proliferation activity of colorectal cancer and paracancerous tissues. The activity of Caspase-3 was detected by kit method and apoptosis was detected by Annexin-V FITC/PI assay. Results the expression of SYK in colorectal cancer tissue and colorectal cancer cell line decreased (P 0.01). The mRNA (P01) and protein expression of SYK were significantly increased (P01) in colorectal cancer cell line pcDNA.3.1-SYK, which showed that the overexpression of SYK was successful. After overexpression of SYK, the growth activity and value-added activity of colorectal cancer cells decreased significantly (P01), and apoptosis increased (P01). In addition, after overexpression of SYK, the expression of Fra-1 was significantly inhibited (P01). Conclusion overexpression of SYK can inhibit the proliferation of colorectal cancer cells and promote the apoptosis of colorectal cancer cells. The mechanism may be related to the regulation of Fra-1 by SYK, which provides a reference value and theoretical basis for the prevention and treatment of colorectal cancer.
【作者单位】： 河南省中医院肛肠科;河南中医药大学中医外科;
【基金】：河南省中医药科学研究专项课题项目(2015ZY02049)
【分类号】：R735.3
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