中国癌症杂志 ›› 2020, Vol. 30 ›› Issue (9): 661-666.doi: 10.19401/j.cnki.1007-3639.2020.09.004

• 论著 • 上一篇    下一篇

节律因子BMAL1通过SHH信号转导通路影响胃癌MGC-803细胞的增殖

程 玉 1 ,郝美玲 2 ,薛 晶 3 ,齐洁敏 1   

  1. 1. 承德医学院病理学教研室,河北 承德 067000 ;
    2. 承德医学院附属医院病理科,河北 承德 067000 ;
    3. 承德医学院形态学实验中心,河北 承德 067000
  • 出版日期:2020-09-30 发布日期:2020-10-10
  • 通信作者: 齐洁敏 E-mail: qijiemin@126.com
  • 基金资助:
    河北省高等学校科学技术研究项目(QN2018130);河北省重点学科建设项目[冀教高(2013)4号病理学与病理生理学]。

BMAL1 influences proliferation of gastric cancer MGC-803 cells through SHH signaling pathway

CHENG Yu 1 , HAO Meiling 2 , XUE Jing 3 , QI Jiemin 1   

  1. 1. Department of Pathology, Chengde Medical College, Chengde 067000, Hebei Province, China; 2. Department of Pathology, Affiliated Hospital of Chengde Medical College, Chengde 067000, Hebei Province, China; 3. Morphology Experimental Center, Chengde Medical College, Chengde 067000, Hebei Province, China
  • Online:2020-09-30 Published:2020-10-10
  • Contact: QI Jiemin E-mail: qijiemin@126.com

摘要: 背景与目的:节律因子BMAL1除了在维持正常生物节律中发挥核心作用外,也参与多种肿瘤的演进过程,但其具体作用机制还未完全阐明。探讨节律因子BMAL1对胃癌细胞增殖的影响及下游信号转导通路。方法:应用靶向BMAL1的小干扰RNA(small interfering RNA,siRNA)进行细胞转染(si-BMAL1组),并设置乱序阴性对照(negative control,NC)组,通过蛋白质印迹法(Western blot)验证细胞转染效率。分别采用MTT法和平板集落形成实验检测细胞的活力和增殖情况,采用Western blot测定细胞中增殖相关功能蛋白cyclin D1的表达水平以及SHH信号转导通路蛋白SHH、SMO、GLI1的表达变化。采用SHH信号通路抑制剂环巴胺处理BMAL1下调后的MGC-803细胞,采用Western blot检测SHH、SMO、GLI1及cyclin D1的蛋白水平,采用平板集落形成实验检测细胞增殖能力的变化。结果:与NC组相比,si-BMAL1组细胞活力增强,细胞中cyclin D1的表达上调,SHH、SMO、GLI1的表达明显上调(P<0.05)。SHH抑制剂环巴胺处理干扰BMAL1的MGC-803细胞后,与si-BMAL1组相比,si-BMAL1+环巴胺组SHH、SMO、GLI1的蛋白水平下调,cyclin D1并未发生明显变化,si-BMAL1促进胃癌细胞增殖的作用可被环巴胺所逆转。结论:BMAL1可能通过SHH信号转导通路抑制胃癌MGC-803细胞的增殖。

关键词: 胃癌, 节律因子, BMAL1, 细胞增殖, SHH信号通路

Abstract: Background and purpose: In addition to its central role in maintaining normal biorhythms, the abnormal expression of rhythm factor BMAL1 may also be involved in the evolution of various tumors. However, the mechanism of action has not been fully elucidated. This study aimed to investigate the effect of BMAL1 on the proliferation of gastric cancer cells and downstream signaling pathways. Methods: si-BMAL1 or negative control (NC) was transfected into the MGC-803 cells, and then Western blot was used to detect the transfection efficiency. The cell viability and proliferation were examined by MTT assay and plate colony formation assay, respectively. The protein level of cyclin D1 and the expressions of SHH signaling pathway-related proteins SHH, SMO and GLI1 in the MGC-803 cells were determined by Western blot. After cyclopamine, a SHH signaling pathway inhibitor was used to treat the gastric cancer MGC-803 cells with BMAL1 knockdown, Western blot was used to detect the protein levels of cyclin D1, SHH, SMO and GLI1. Plate colony formation assay was used to measure the viability of the cells. Results: Compared with NC group,the expression of BMAL1 in si-BMAL1 group was significantly down-regulated,and the cell proliferation was increased. The protein levels of cyclin D1, SHH, SMO and GLI1 in the cells were increased (P<0.05). After treatment with SHH inhibitor cyclopamine, the protein levels of SHH, SMO and GLI1 were lower in si-BMAL1+cyclopamine group than in NC group, while cyclin D1 did not change. SHH signaling pathway inhibitor reversed BMAL1 knockdown-induced increase in viability of MGC-803 cells. Conclusion: BMAL1 inhibits the growth of gastric cancer cells through SHH signaling pathway.

Key words: Gastric cancer, Rhythm factor, BMAL1, Cell proliferation, SHH signaling pathway