主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2016, Vol. 39 ›› Issue (11): 909-914.doi: 10.3969/j.issn.1006-2157.2016.11.005

• 科技之窗 • 上一篇    下一篇

健脾益气化瘀解毒方含药血清对结肠癌HCT116细胞增殖、周期、凋亡的影响

简小兰1,杨晓1,罗吉2,李勇敏2,吕元2,谭小宁2,蒋益兰2#   

  1. 1 湖南中医药大学研究生院 湖南 410208;
    2 湖南省中医药研究院附属医院
  • 收稿日期:2016-06-03 出版日期:2016-11-30 发布日期:2016-11-30
  • 通讯作者: 蒋益兰,女,主任医师,博士生导师,研究方向:肿瘤中西医结合防治方法和规律研究,E-mail:tianshangren624@163.com
  • 作者简介:简小兰,女,在读博士生
  • 基金资助:
    国家自然科学基金资助项目(No.81373602),湖南省研究生科研创新项目(No.CX2015B319),湖南省中医药研究院项目(No.201504)

Effect of Jianpi Yiqi Huayu Jiedu Formula on proliferation, cycle and apoptosis of HCT116 colorectal cancer cells

JIAN Xiaolan1, YANG Xiao1, LUO Ji2, LI Yongmin2, LYU Yuan2, TAN Xiaoning2, JIANG Yilan2#(1 Graduate School of Hunan University of Chinese Medicine, Hunan 410208; 2 The Affiliated Hospital of Hunan Academy of Chinese Medicine)   Abstract: Objective To study the effect of medicated serum of Jianpi Yiqi Huayu Jiedu Formula   

  1. 1 Graduate School of Hunan University of Chinese Medicine, Hunan 410208;
    2 The Affiliated Hospital of Hunan Academy of Chinese Medicine
  • Received:2016-06-03 Online:2016-11-30 Published:2016-11-30

摘要: 目的 探讨健脾益气化瘀解毒方含药血清对人结肠癌HCT116细胞增殖、周期、凋亡及半胱氨酸天冬氨酸蛋白酶-3(Caspase-3),凋亡相关蛋白Bcl-xL和Bax蛋白表达的影响。方法 10%、15%、20%健脾益气化瘀解毒方含药血清处理HCT116细胞12、24、48、72 h后,MTT法检测细胞增殖;不同浓度含药血清处理24、48 h后,碘化丙啶染色法检测细胞周期及凋亡率。蛋白质印迹法(Western blot)检测HCT116细胞内Caspase-3,Bcl-xL和Bax蛋白的表达;对照组均予以相同浓度空白鼠血清及10%胎牛血清(FBS)。结果 细胞增殖:12 h后15%、20%含药血清与相同浓度鼠空白组比较,对HC116细胞的抑制作用具有统计学意义(P<0.05),72 h对HCT116细胞的抑制作用最强(P<0.05),健脾益气化瘀解毒方含药血清对HCT116细胞增殖具有良好的抑制作用,并与时间、浓度呈正相关。细胞周期:10%及15%含药血清组G2期细胞减少,而G1期与S期细胞增多,20%含药血清组G1期与S期细胞减少,而G2期细胞增加,48 h作用明显,而空白鼠血清细胞周期则无明显变化,说明含药血清能阻滞细胞周期,与时间呈正相关。细胞凋亡:与相应浓度对照组比较,含药血清处理24、48 h后,HCT116细胞晚期凋亡率增加,并且随浓度、时间增加凋亡率增加(P<0.05),说明含药血清能促使HCT116细胞凋亡,与时间、浓度呈正相关。Western blot检测显示,与空白鼠血清组相比,15%含药血清可上调Bax蛋白表达,下调Caspase-3、Bcl-xL的蛋白表达(P<0.05)。结论 健脾益气化瘀解毒方可能通过将结肠癌HCT116细胞周期阻滞,从而抑制细胞增殖,促进细胞凋亡,通过上调凋亡相关蛋白Bax,下调Caspase-3、Bcl-xL蛋白的表达诱导细胞凋亡。

关键词: 健脾益气化瘀解毒, HCT116细胞, 增殖, 细胞周期, 凋亡

Abstract: Objective To study the effect of medicated serum of Jianpi Yiqi Huayu Jiedu Formula (JYHJF), a TCM compound formulated according to TCM principle of tonifying spleen, replenishing qi, removing blood stasis and detoxification, on proliferation, cycles, apoptosis of HCT116 colorectal cancer cells and on the protein expression of Caspase-3,Bcl-xL and Bax of HCT116 cells. Methods After drug-contained serum (10%, 15%, 20%) treated HCT116 cells for 12, 24, 48 and 72 h, the proliferation was detected by MTT method; after treated for 24 or 48 h, the cell cycle and apoptosis rate were detected by propidium iodide (PI) staining. The protein expression level of Caspase 3, Bcl-xL and Bax in HCT116 cells were detected by Western blot. The control group was treated by blank serum and 10% fetal bovine serum (FBS). Results Compared with the same concentration of blank serum group, the cell proliferation was inhibited significantly in 15% and 20% medicated serum group after 12 h (P<0.05), and the inhibitory effect showed time- and concentration- dependence with the highest at 72 h (P<0.05). In 10% and 15% medicated serum group, cells in G2 phase decreased, but the G1-phase cells and S-phase cells increased; cells in G1 phase and S phase increased and those in G2 phase decreased in 10% medicated serum group with the remarkable Results at 48 h. It was suggested that the medicated serum could blocked cell cycles, with the positive correlation with time. Compared with the blank serum groups at the same concentration, after treated 24 and 48 h, apoptosis rate of HCT116 cells increased, and had the positive correlation with time and concentration. Compared with the blank serum group, the expression of Bax protein was upregulated and that of Caspase-3 and Bcl-xL were downregulated in 15% medicated serum group (P<0.05). Conclusion The effect of Jianpi Yiqi Huayu Jiedu Formula on inhibiting proliferation and inducing apoptosis may be via the way of blocking HCT116 cells cycles and of upregulating Bax protein expression with downregulating Caspase-3 protein and Bcl-xL protein expression.

Key words: Jianpi Yiqi Huayu Jiedu Formula , HCT116 colorectal cancer cells , proliferation, cell cycle, apoptosis

中图分类号: 

  • R285.5