肝癌全方及其不同治法拆方抑制SMMC7721人肝癌细胞增殖的作用

王晓敏; 潘志强; 梁龙龙; 方肇勤

doi:10.13422/j.cnki.syfjx.20181116

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肝癌全方及其不同治法拆方抑制SMMC7721人肝癌细胞增殖的作用

药理 | 更新时间：2024-01-04

- 肝癌全方及其不同治法拆方抑制SMMC7721人肝癌细胞增殖的作用
- Inhibitory Effect of Hepatocellular Carcinoma Prescription and Its Different Disassembled Prescriptions on Proliferation of SMMC7721 Hepatoma Cells
- 中国实验方剂学杂志 2018年24卷第13期页码：117-123
- 作者机构：
- 作者简介：
- 基金信息：
  
  国家自然科学基金面上项目（81473562）
- DOI：10.13422/j.cnki.syfjx.20181116
  中图分类号： R285.5
- 纸质出版日期：2018
- 稿件说明：
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王晓敏, 潘志强, 梁龙龙, 等. 肝癌全方及其不同治法拆方抑制SMMC7721人肝癌细胞增殖的作用[J]. 中国实验方剂学杂志, 2018,24(13):117-123.

WANG Xiao-min, PAN Zhi-qiang, LIANG Long-long, et al. Inhibitory Effect of Hepatocellular Carcinoma Prescription and Its Different Disassembled Prescriptions on Proliferation of SMMC7721 Hepatoma Cells[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(13): 117-123.
王晓敏, 潘志强, 梁龙龙, 等. 肝癌全方及其不同治法拆方抑制SMMC7721人肝癌细胞增殖的作用[J]. 中国实验方剂学杂志, 2018,24(13):117-123. DOI： 10.13422/j.cnki.syfjx.20181116.

WANG Xiao-min, PAN Zhi-qiang, LIANG Long-long, et al. Inhibitory Effect of Hepatocellular Carcinoma Prescription and Its Different Disassembled Prescriptions on Proliferation of SMMC7721 Hepatoma Cells[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(13): 117-123. DOI： 10.13422/j.cnki.syfjx.20181116.

摘要

目的：探讨肝癌全方及不同治法拆方抗肿瘤作用的分子机制。方法：体外培养SMMC7721人肝癌细胞，分别给予2.5～100 g·L-1肝癌全方、清热方、活血方、健脾方干预24 h后，采用噻唑蓝（MTT）比色法检测细胞增殖；分别采用1.25～20 g·L-1肝癌全方，5～20 g·L-1清热方和活血方及20～120 g·L-1健脾方干预24 h后，运用实时荧光定量聚合酶链式反应（Real-time PCR）检测相关基因的表达；分别采用20 g·L-1肝癌全方和清热方，10 g·L-1活血方及120 g·L-1健脾方干预SMMC7721细胞24 h后，通过蛋白免疫印迹法检测相关蛋白的表达，通过碘化丙啶（PI）/RNase staining solution试剂染色检测细胞周期的改变。结果：与空白组比较，肝癌全方及其不同治法拆方能抑制SMMC7721细胞的增殖，并呈现量效关系（P<0.05）。与空白组比较，5~20 g·L-1清热方呈现量效抑制细胞周期依赖性激酶阻滞基因1B（CDKN1B）基因表达（P<0.05），20~120 g·L-1健脾方也呈现量效抑制CDKN1B基因表达（P<0.05）；5~20 g·L-1肝癌全方明显抑制糖皮质激素调节蛋白激酶1（SGK1）基因表达（P<0.05），且量效显著；5~20 g·L-1清热方量效抑制SGK1基因表达（P<0.05）；20~120 g·L-1健脾方也抑制SGK1基因表达（P<0.05），同时肝癌全方及清热、活血治法方可抑制SGK1磷酸化蛋白的表达（P<0.05）；1.25，20 g·L-1肝癌全方可促进叉头框蛋白O3（FOXO3）基因表达（P<0.05）；15，20 g·L-1清热方明显促进FOXO3基因表达（P<0.05）；20 g·L-1活血方促进FOXO3基因表达（P<0.05）。与空白组比较，全方组G2期细胞数增加（P<0.05），清热组和健脾组G2期细胞数增加更明显（P<0.05）；肝癌全方及不同治法拆方能够抑制细胞周期蛋白E1（CyclinE1）蛋白的表达，其中以健脾组抑制效果最显著。结论：肝癌全方及其不同治法拆方通过改变细胞增殖和周期，从而发挥抑制肝癌的作用。

Abstract

Objective: To study the molecular mechanism on anti-tumor effect of hepatocellular carcinoma prescription (Quanfang) and its different disassembled prescriptions. Method: SMMC7721 human hepatoma cells were cultured in vitro and treated with 2.5-100 g·L-1 Quanfang

Qingrefang

Huoxuefang and Jianpifang for 24 h. Then cells proliferation was measured by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay. After SMMC7721 cells were treated with 1.25-20 g·L-1 Quanfang

5-20 g·L-1 Qingrefang and Huoxuefang and 20-120 g·L-1 Jianpifang for 24 h

related gene expression levels were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). After SMMC7721 cells were treated with 20 g·L-1 Quanfang and Qingrefang

10 g·L-1 Huoxuefang and 120 g·L-1 Jianpifang for 24 h

related protein expression levels were detected by Western blot and cell cycle changes were detected by PI/RNase staining solution reagent staining. Result: As compared with blank control group

Quanfang and different disassembled prescriptions inhibited the proliferation of SMMC7721 cells and showed dose-effect relationship (P<0.05). As compared with the blank control group

CDKN1B gene expression was inhibited significantly by 5-20 g·L-1 Qingrefang and 20-120 g·L-1 Jianpifang (P<0.05) in a dose-effect manner. SGK1 gene expression was inhibited significantly by 5-20 g·L-1 Quanfang and Qingrefang (P<0.05); Quanfang and different disassembled prescriptions can inhibited SGK1 phosphorylated protein expression. SGK1 gene expression was inhibited by 5-20 g·L-1 Qingre disassembled prescriptions and 20-120 g·L-1 Jianpi disassembled prescriptions (P<0.05). FOXO3 gene expression was promoted significantly respectively by 1.25

20 g·L-1 Quanfang

20 g·L-1 Qingre disassembled prescriptions and 20 g·L-1 Huoxue disassembled prescriptions (P<0.05). As compared with blank control group

the number of G2/M cells was increased in Quanfang group (P<0.05) and was increased more significantly in Qingre group and Jianpi group (P<0.05). CyclinE1 protein expression was inhibited by Quanfang and different disassembled prescriptions

and the effect was most significant in Jianpi group. Conclusion: Quanfang and different disassembled prescriptions play a role in inhibiting liver cancer by altering cell proliferation and cycle.

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