杨晓　唐蔚　蒋益兰

摘要：目的 观察健脾消癌方对Wnt/β-catenin信号通路相关因子及人结肠癌HCT116细胞周期、凋亡的影响，探讨其抗大肠癌复发转移的作用机制。方法 将对数生长期的HCT116细胞分为空白组、生理盐水组、5-Fu组及健脾消癌方低、中、高剂量组，分别干预48 h，收集细胞，采用流式细胞术检测细胞周期、凋亡，Western blot检测细胞核内β-catenin蛋白表达，PCR检测c-myc、cyclinD1 mRNA表达。结果 与空白组及生理盐水组比较，健脾消癌方各剂量组HCT116细胞凋亡比例、G1期细胞比例增加，S期细胞比例降低，且其细胞核内β-catenin蛋白表达降低，c-myc、cyclinD1 mRNA表达降低，尤以健脾消癌方高剂量组明显，差异有统计学意义（P<0.05，P<0.01）。结论 健脾消癌方可促进人结肠癌HCT116细胞凋亡并将其细胞周期阻滞，其机制可能与调节Wnt/β-catenin信号通路有关。

关键词：健脾消癌方；大肠癌；Wnt/β-catenin信号通路；细胞凋亡；细胞周期

DOI：10.3969/j.issn.1005-5304.2018.06.015

中圖分类号：R285.5 文献标识码：A 文章编号：1005-5304（2018）06-0061-05

Effects of Jianpi Xiaoai Prescription on Cell Cycle and Apoptosis of Human Colon Cancer HCT116 Cells and Wnt/β-catenin Signaling Pathway Related Proteins

YANG Xiao1， TANG Wei2， JIANG Yi-lan2， TAO Zi-hao1， LIU Ke1， SONG Cheng3

1. Graduate School of Hunan University of Chinese Medicine， Changsha 410208， China；

2. The Affiliated Hospital of Hunan Academy of Chinese Medicine， Changsha 410006， China；

3. Hunan Cancer Hospital， The Affiliated Cancer Hospital of Xiangya School of Medicine，

Central South University， Changsha 410006， China

Abstract： Objective To study the effects of Jianpi Xiaoai Prescription on cell cycle and apoptosis of human colon cancer HCT116 cells and related factors of Wnt/β-catenin signaling pathway； To investigate its mechanisms of anti-metastasis and recurrence of colorectal cancer. Methods The logarithmic growth phase HCT116 cells were divided into blank group， saline group， 5-Fu group， and Jianpi Xiaoai Prescription low-， medium-， and high-dose groups. After intervention for 48 h， the cells were harvested， and the cell cycle and apoptosis were detected by flow cytometry. The protein expression of β-catenin in the nucleus was detected by Western blot， and the expression of c-myc and cyclinD1 mRNA was detected by PCR. Results Compared with the blank group and saline group， the ratio of HCT116 cells apoptosis of Jianpi Xiaoai Prescription low-， medium-， and high-dose groups increased； the proportion of cells in phase G1 increased and the proportion of S cells decreased； the expression of β-catenin protein in the nucleus and the expression of c-myc， cyclinD1 mRNA decreased， especially in the Jianpi Xiaoai Prescription high-dose group， with statistical significance （P<0.05， P<0.01）. Conclusion Jianpi Xiaoai Prescription can promote apoptosis of human colon cancer HCT116 cells and block the cell cycle， and its mechanism may be related to regulating Wnt/β-catenin signaling pathway.

Keywords： Jianpi Xiaoai Prescription； colorectal cancer； Wnt/β-catenin signaling pathway； apoptosis； cell cycle

大肠癌（colorectal cancer，CRC）指发生于结肠及直肠的恶性肿瘤。近年来，随着生活方式及饮食结构的改变，我国CRC发病率急剧上升。目前，CRC的治疗仍以外科手术为主，辅以化学药物、放射、靶向药物等治疗，术后5年生存率约50%。近年来，本课题组运用中药复方健脾消癌方治疗结直肠癌患者已达1万余人次，临床疗效较好[1-3]，但其作用机理尚不明确。研究表明，Wnt信号通路可通过影响细胞凋亡、增殖和分化等生命活动参与结直肠癌的发生和发展[4]，大约90%结直肠癌患者存在Wnt信号通路的异常[5]。本实验以人结肠癌HCT116细胞为实验对象，研究健脾消癌方对CRC细胞凋亡及Wnt/β-catenin信号通路相关因子的影响，探讨其抗CRC复发转移的作用机制。

1 实验材料

1.1 动物

SPF級雄性健康Wistar大鼠15只，4～6周龄，体质量（300±20）g，长沙斯莱克景达实验动物有限公司，动物合格证号SCXK（湘）2016-0002，饲养于湖南省中医药研究院动物房，温湿度适宜，人工12 h昼/夜循环照明，灌胃前适应性饲养3 d。……