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目的:探讨维生素D3在结肠癌中的抗肿瘤机制及其与抑癌基因runt相关转录因子3(RUNX3)的关系,为临床有效治疗结肠癌提供依据。方法:不同浓度维生素D3作用于SW480结肠癌细胞系,RT-PCR和Western blot分别检测RUNX3在mRNA和蛋白水平的表达情况。构建RUNX3基因的小干扰RNA载体并将其转染SW480结肠癌细胞,经G418筛选获得稳定表达的细胞系。MTT法和流式细胞术检测维生素D3对转染前后的细胞生长作用的影响。结果:随着维生素D3作用浓度增加,SW480细胞中RUNX3基因在mRNA和蛋白表达水平均呈剂量依赖性增加。成功构建了RUNX3的小干扰RNA载体并将其转染SW480细胞;筛选到稳定的敲除RUNX3的结肠癌细胞系。与其他对照组相比,维生素D3对RUNX3敲除的SW480细胞的生长抑制作用减弱。结论:维生素D3在转录和翻译水平正性调控RUNX3的表达而抑制结肠癌细胞的增殖。
Objective: To investigate the anti-tumor mechanism of vitamin D3 in colon cancer and its relationship with runt-associated transcription factor 3 (RUNX3), so as to provide a basis for clinical effective treatment of colon cancer. Methods: Different concentrations of vitamin D3 were applied to SW480 colon cancer cell lines. The expression of RUNX3 mRNA and protein were detected by RT-PCR and Western blot respectively. Small interfering RNA vector of RUNX3 gene was constructed and transfected into SW480 colon cancer cells, and stable cell line was obtained by G418 screening. MTT assay and flow cytometry were used to detect the effect of vitamin D3 on cell growth before and after transfection. Results: With the increase of vitamin D3 concentration, RUNX3 mRNA and protein expression in SW480 cells increased in a dose-dependent manner. The small interfering RNA vector of RUNX3 was successfully constructed and transfected into SW480 cells; a stable colon cancer cell line that knocked out RUNX3 was screened out. Compared with other control groups, vitamin D3 inhibited the growth inhibition of RUNX3 knockout SW480 cells. Conclusion: Vitamin D3 can inhibit the proliferation of colon cancer cells by regulating the expression of RUNX3 at the transcription and translation levels.