藻蓝色素蛋白下调RIPK1表达并抑制肺癌A549细胞的增殖和迁移

C-phycocyanin Inhibits the Proliferation of Non-small Cell Lung Cancer A549 Cells by Down-regulating RIPK1 Expression

以非小细胞肺癌A549细胞为模型,探究藻蓝蛋白抑制其增殖和迁移的调控机制。采用MTT法、克隆形成试验、流式细胞术、划痕试验等方法检测藻蓝色素蛋白对细胞增殖、迁移的影响。采用转录组学测序分析筛选关键基因:受体相互作用蛋白激酶1(Receptor interacting proteinkinase1,RIPK1),通过体外转染小干扰RNA沉默RIPK1,利用荧光定量PCR、免疫印迹技术检测沉默效率,验证沉默RIPK1后细胞增殖机制及体外迁移能力的变化。结果表明,藻蓝蛋白降低了细胞的存活率、生长速率、集落形成能力以及体外迁移能力。细胞转染RIPK1 siRNA后,RIPK1的表达受到显著抑制,同时,细胞生长速率、集落形成能力以及体外迁移能力也降低,并且细胞周期出现G1期阻滞,这与藻蓝蛋白处理A549细胞的表型一致。本研究证实了藻蓝蛋白能够通过下调RIPK1表达抑制肺癌A549细胞的增殖和迁移,为利用新型抗癌功能性食品着色剂奠定了基础,同时也对非小细胞肺癌的安全性治疗提供了理论依据。

In this study,non-small cell lung cancer A549 was used as a model to determine the mechanism by which C-phycocyanin(C-PC)regulated the biological behavior of A549 cells.MTT assay,clonal formation assay,flow cytometry and scratch assay were used to examine the effects of C-PC on cell proliferation and migration.The key gene receptor interacting protein kinase 1(RIPK1)was screened by transcriptome sequencing analysis,the silencing efficiency was detected by real-time PCR and western blot,and the effect of cell proliferation and migration capacity in vitro was verified by transfecting RIPK1 small interfering RNA.The C-PC reduced cell survival rate,growth rate,and cell colony formation ability.After transfecting RIPK1 siRNA,the expression level of RIPK1 was inhibited.Meanwhile,cell growth rate,colony formation and migration ability were suppressed.The cell cycle exerted G1 phase arrest.This work indicated that C-PC could inhibit the proliferation and migration of A549 cells by down-regulating RIPK1 expression level,which laid a foundation for the deep utilization of new anti-cancer functional food colorants.It also provides the necessary theory for the safe treatment of non-small cell lung cancer.