补肾解毒散结方醇提物调控HIPK2-p53信号通路抑制人结肠癌细胞增殖

Bushen Jiedu Sanjie Recipe alcohol extract inhibit cell proliferation through HIPK2-p53 signaling pathway in colorectal cancer

目的:探讨补肾解毒散结方调控HIPK2-p53信号通路抗人结肠癌细胞增殖的作用和机制。方法:裸鼠腋下接种人结肠癌HCT-116细胞,建立人大肠癌(CRC)裸鼠移植瘤模型,随机分为模型组,补肾解毒散结方低、中、高剂量组,并与5-氟尿嘧啶(5-FU)组做对照,连续给药2周,之后处死裸鼠,比较各组裸鼠瘤体体积及相对抑瘤率;以不同浓度的补肾解毒散结方醇提物(10、20、40、60、80、100、120μg/m L)分别作用于人结肠癌HCT-116细胞48h,CCK-8法检测补肾解毒散结方对HCT-116细胞增殖的抑制作用;单克隆形成实验分析补肾解毒散结方对HCT-116细胞克隆形成情况;Western blot法检测补肾解毒散结方对细胞HIPK2、p53蛋白表达的影响。结果:补肾解毒散结方低、中、高剂量组及5-FU组对裸鼠瘤体大小抑制率分别为28.83%、39.98%、47.67%和54.26%;补肾解毒散结方对结肠癌HCT-116细胞的增殖有明显的抑制作用,并呈剂量依赖性,48h的半数抑制浓度(IC50)值为57.59μg/m L;补肾解毒散结方对HCT-116细胞的克隆形成有明显的抑制作用,呈剂量依赖性;补肾解毒散结方作用HCT-116细胞48h后,剂量依赖性地上调HIPK2、p53蛋白表达。结论:补肾解毒散结方能抑制人结肠癌HCT-116细胞增殖及人结肠癌裸鼠移植瘤瘤体生长,其机制可能与上调HIPK2、p53的表达从而抑制肿瘤生长相关。

Objective：To investigate the the impact and mechanism of Bushen Jiedu SanJie Recipe（BSF） inhibit the proliferation through HIPK2-p53 signaling pathway in colorectal cancer cell line HCT-116.Methods：Nude mouse model of colorectal HCT-116 cell cancer was established,and randomly divided into model group,BSF low,medium,high dose and 5-FU groups.After 2 weeks treatment with medicine respectively,the mice in each group were measure tumor tissues sizes,inhibition rate;Various concentrations of BSF alcohol extract were set as 10,20,40,60,80,100 and 120ug/mL,and then the proliferation of CRC HCT-116 cells was examined using CCK-8;The formation of the clone of HCT-116 cells was analyzed by the experiment of the monoclonal formation;The expression of HIPK2 and p53 with effect BSF was detected by Western blot.Results：The inhibition rates of the low,medium,high dose and 5-FU on the tumor volume of BSF,respectively were 28.83%,39.98%,47.67%and 54.26%,BSF significantly inhibited proliferation of CRC cell lines HCT-116,in a dose-dependent manner.The half inhibition rates were 57.59μg/mL in 48 h,BSF significantly inhibited monoclonal formation of CRC cell lines HCT-116,in a dose-dependent manner.BSF treatment significantly enhanced the protein expression of HIPK2,p53 in CRC cells.Conclusion：BSF significantly inhibited proliferation of CRC cell lines HCT-116 and human colon cancer in nude mice transplanted tumor growth,via a probable mechanism of enhancing HIPK2 and p53 expression.