藤梨根提取物通过调控miR-192-5p/ARPP19轴影响结直肠癌细胞的增殖和凋亡  

Radix Actinidiae extract affects proliferation and apoptosis of colorectal cancer cells by regulating miR-192-5p/ARPP19

作　　者：徐万苏[1] 柯飞[2] 许怡 郑艺[3] Wan-Su Xu;Fei Ke;Yi Xu;Yi Zheng(Department of Oncology Radiotherapy,Quzhou People’s Hospital,Quzhou 324000,Zhejiang Province,China;Department of Pathology,Quzhou People’s Hospital,Quzhou 324000,Zhejiang Province,China;Sterilization Supplying Center,Quzhou People’s Hospital,Quzhou 324000,Zhejiang Province,China)

机构地区：[1]衢州市人民医院肿瘤放疗科,浙江省衢州市324000 [2]衢州市人民医院病理科,浙江省衢州市324000 [3]衢州市人民医院消毒供应中心,浙江省衢州市324000

出　　处：《世界华人消化杂志》2021年第8期398-406,共9页World Chinese Journal of Digestology

摘　　要：背景藤梨根提取物(rattan root extract,RRE)可抑制胃癌、肺癌等肿瘤细胞增殖,具有一定抗肿瘤作用,但其是否影响结直肠癌细胞的恶性表型还未知.miR-192-5p在结直肠癌组织中表达降低,且其低表达与肿瘤大小等临床病理特征相关,可作为结直肠癌诊治的潜在生物学标志物.StarBase生物信息学软件预测显示,环腺苷酸调节的磷酸化蛋白19(cAMP-regulated phosphoprotein 19,ARPP19)可能是miR-192-5p的靶基因.本研究假设RRE可通过调控miR-192-5p/ARPP19轴影响结直肠癌细胞增殖和凋亡.目的探讨RRE对结直肠癌SW480细胞增殖和凋亡的影响及作用机制.方法RRE干预SW480细胞后,MTT检测细胞增殖,流式细胞术检测细胞凋亡,蛋白质印迹法检测细胞中CyclinD1、C-caspase-3和ARPP19蛋白水平,RT-qPCR检测细胞中miR-192-5p和ARPP19 mRNA水平.转染miR-192-5p模拟物或ARPP19小干扰RNA至SW480细胞,上述相同方法观察过表达miR-192-5p或抑制ARPP19表达对SW480细胞增殖、凋亡及CyclinD1和C-caspase-3蛋白表达的影响.双荧光素酶报告基因实验验证miR-192-5p和ARPP19调控关系.结果RRE可降低SW480细胞存活率及ARPP19 mRNA和蛋白的表达(P<0.05),增加SW480细胞凋亡率及C-caspase-3蛋白和miR-192-5p的表达(P<0.05).过表达miR-192-5p或抑制ARPP19表达均可降低SW480细胞存活率及CyclinD1蛋白表达(P<0.05),而提高SW480细胞凋亡率及C-caspase-3蛋白表达(P<0.05).miR-192-5p在SW480细胞中靶向负调控ARPP19表达.抑制miR-192-5p表达可逆转RRE对SW480细胞增殖、凋亡及CyclinD1和C-caspase-3蛋白表达的影响.抑制ARPP19表达可逆转抑制miR-192-5p表达对RRE处理的SW480细胞增殖、凋亡及CyclinD1和C-caspase-3蛋白表达的影响.结论RRE可有效抑制结直肠癌SW480细胞增殖,促进细胞凋亡,其作用机制与调miR-192-5p/ARPP19轴有关.BACKGROUND Radix Actinidiae extract(RAE)can inhibit the proliferation of gastric cancer,lung cancer,and other tumor cells,and has appreciated anti-tumor effects,but it is unknown whether it affects the malignant phenotype of colorectal cancer cells.The expression of miR-192-5p is reduced in colorectal cancer tissues,and its low expression is related to clinicopathological characteristics such as tumor size.Thus,it can be used as a potential biomarker for the diagnosis and treatment of colorectal cancer.StarBase bioinformatics software predicts that cAMP-regulated phosphoprotein 19(ARPP19)may be a target gene of miR-192-5p.This study hypothesized that RAE can affect the proliferation and apoptosis of colorectal cancer cells by regulating the miR-192-5p/ARPP19 axis.AIM To investigate the effects and mechanism of RAE on proliferation and apoptosis of colorectal cancer SW480 cells.METHODS After SW480 cells were treated with RAE,cell proliferation was detected by MTT assay,apoptosis was detected by flow cytometry,and the protein expression levels of Cyclin D1,cleaved Caspase-3,and ARPP19 were detected by Western blot.The expression of miR-192-5p and ARPP19 mRNA in cells was detected by RTqPCR.miR-192-5p mimic or ARPP19 small interfering RNA was transfected into SW480 cells,and then the above methods were used to observe the effects of overexpressing miR-192-5p or inhibiting ARPP19 on cell proliferation,apoptosis,and the protein expression of CyclinD1 and cleaved Caspase-3 in SW480 cells.The regulatory relationship between miR-192-5p and ARPP19 was verified by the dual luciferase reporter gene assay.RESULTS RAE decreased the survival rate of SW480 cells and the expression of ARPP19 mRNA and protein(P<0.05),but increased the apoptosis rate of SW480 cells and the expression of cleaved Caspase-3 protein and miR-192-5p(P<0.05).Over-expressing miR-192-5p or inhibiting ARPP19 expression could decrease the survival rate of SW480 cells and the expression of CyclinD1 protein(P<0.05),but increase the apoptosis rate of SW480 cell

关 键 词：结直肠癌 藤梨根提取物 miR-192-5p 环腺苷酸调节的磷酸化蛋白19 细胞增殖 凋亡 

分 类 号：R285.5[医药卫生—中药学]