摘要
目的：研究通莲汤对胃癌MGC803细胞形态、生长周期的调节作用及细胞核转录因子κB（NF-κB）信号通路的影响,以探讨该方抑制胃癌细胞增殖的机制.方法：将MGC803细胞以1×104个细胞/孔的密度接种于96孔细胞培养板,以含5％胎牛血清DMEM培养液培养,37℃,5％饱和湿度的CO2培养箱孵育,细胞贴壁24 h,分别加入浓度为25～800 mg·L-倍比梯度的通莲汤提取液,以10 ～ 160 mg·L-1倍比浓度的氟脲嘧啶为阳性对照组,孵育48 h后,倒置显微镜下观察细胞形态,MTT比色法检测细胞增殖变化,曲线拟合法计算通莲汤和氟脲嘧啶50％抑制浓度（IC50）;以两者48 h IC50浓度处理细胞48 h,流式细胞技术（FCM）测定细胞周期,Western blot法测定细胞IκB激酶β（IKKβ）,NF-κB,phosph-NF-κB蛋白水平.结果：通莲汤和氟脲嘧啶48 h IC50值分别为192.74 mg.L-1,23.61 mg·L-1;与空白对照组相比,通莲汤显著影响MGC803细胞周期中G2期,氟脲嘧啶显著影响MGC803细胞周期中G1期（P＜0.05）;通莲汤和氟脲嘧啶均能显著抑制IKKβ与NF-κB的蛋白表达.结论：通莲汤抑制MGC803细胞生长与抑制细胞于G2期,下调NF-κB通路中IKKβ,NF-κB蛋白的表达有关.
Objective：To observe the effects of Tonglian decoction （TD） of morphology,the growth cycle and nuclear factor-kappa B （NF-κB） signaling pathway,and to discuss the inhibiting mechanisms on gastric cancer cell proliferation in gastric carcinoma cell line MGC803.Method：MGC803 cell line was seeded in 96-well micro-plate with 1 × 104 cells in each well,and cultured with DMEM containing 5% fetal bovine serum （FBS）,and maintained in the incubator at 37 ℃ with 5% CO2 in humidified atmosphere.After cell adherence for 24 h,TD of different concentrations was administrated from 25-800 mg ·L-1 with common ration of two,while Fluorouracil from 10-160 mg·L-1 as positive control group.48 hours＇ administration was followedby observation on cell morphology by inverted microscope,and cell proliferation determined by MTT assay.The half maximum inhibition rates （IC50） of TD and fluorouracil were calculated by curve fitting method.After 48 hours＇ administration of TD and Fluorouracil at 48 h IC50 concentration,flow cytometry （FCM） was employed for cell cycle determination,and western blot for protien expression of inhibitor kappa B kinaseβ （IKKβ）,NF-κB and phosph-NF-κB.Result：48h IC50 of TD is 192.74 mg ·L-1,and that of Fluorouracil is 23.61 mg ·L 1.Compared with control group,TD can significantly affect G2 phase of cell cycle in cell line MGC803,and Fluorouracil G1 phase （P 〈 =.05） ; TD and fluorouracil could significantly decrease protein expression of IKKβ and NF-κB.Conclusion：TD has inhibition effect on cell line MGC803,which is related to inhibiting cells in G2 phase,and down-regulating protein expression of IKKβ and NF-κB.
出处
《中国实验方剂学杂志》
CAS
北大核心
2013年第22期213-217,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
深圳市科技研发资金计划-国际科技合作研发项目(ZYA201007060089A)
