G蛋白信号调节因子-13通过调控β-catenin抑制结直肠癌细胞增殖、迁移和侵袭

Regulator of G protein signaling 13 plays an inhibitory role in the progression of colorectal cancer via regulatingβ-catenin pathway

目的:探究G蛋白信号调节因子-13(RGS13)在结直肠癌(CRC)进展中的作用。方法:使用TCGA数据库和实时荧光定量聚合酶链反应(RT-qPCR)在mRNA水平分析CRC组织和细胞中RGS13的表达量,使用免疫组织化学染色(IHC)和蛋白质印迹法(Western blot)在蛋白水平进一步分析。用ATP细胞活力检测实验、软琼脂克隆集落形成实验和细胞迁移侵袭实验检测RGS13对CRC细胞增殖、迁移和侵袭的影响,并通过Western blot、RT-q PCR等实验探究其下游分子机制。结果:RGS13在CRC组织与细胞系中低表达(P<0.01),RGS13表达越低,患者的无病生存期越短(P=0.017)。RGS13的下调可显著促进CRC细胞的增殖、迁移与侵袭(P<0.01),表明RGS13在CRC进展中发挥抑癌作用。机制上,RGS13通过下调Wnt/β-catenin信号通路中β-catenin的蛋白水平,进而降低癌基因c-Myc、MMP7和CCND1等的表达水平(P<0.01),发挥对CRC的抑制作用。结论:RGS13可能通过下调β-catenin发挥抑制CRC进展的重要作用。

Objective:To explore the role of regulator of G protein signaling 13(RGS13)in the progression of colorectal cancer(CRC).Methods:The expression of RGS13 in CRC tissues and cells was analyzed at the mRNA level using the TCGA database and real-time quantitative polymerase chain reaction(qPCR).Further analysis at the protein level was performed using immunohistochemistry(IHC)and western blot.The effect of RGS13 on the proliferation,migration and invasion of CRC cells was detected by ATP cell viability assay,soft agar colony formation assay and cell migration and invasion assay.The downstream molecular mechanism was explored by western blot,RT-qPCR and other experiments.Results:The expression of RGS13 was low in CRC tissues and cell lines(P<0.01).The lower the expression of RGS13,the shorter the disease-free survival of patients(P=0.017).Down-regulation of RGS13 could significantly promote the proliferation,migration and invasion of CRC cells(P<0.01),indicating that RGS13 played a tumor suppressor role in the progression of CRC.Mechanistically,RGS13 inhibited CRC by down-regulating the protein level ofβ-catenin in the Wnt/β-catenin signaling pathway,thereby reducing the expression levels of oncogenes c-Myc,MMP7 and CCND1(P<0.01).Conclusion:RGS13 may play an important role in inhibiting CRC progression by down-regulatingβ-catenin.