启膈散与顺铂对食管癌细胞EC9706低氧下生长抑制及其miR-21,PDCD4和PTEN表达的影响

杨联河; 赵雪艳; 郭彬; 牛春玲; 吴耀松; 陈玉龙; 尹素改; 任闪闪

doi:10.13422/j.cnki.syfjx.2016150134

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启膈散与顺铂对食管癌细胞EC9706低氧下生长抑制及其miR-21,PDCD4和PTEN表达的影响

药理 | 更新时间：2024-09-23

- 启膈散与顺铂对食管癌细胞EC9706低氧下生长抑制及其miR-21,PDCD4和PTEN表达的影响
- Inhibitive Effect of Qige San and Cisplatin on Growth of Esophageal Carcinoma Cells EC9706 and Its miR-21, PDCD4 and PTEN Expressions Under Hypoxia
- 中国实验方剂学杂志 2016年22卷第15期页码：134-138
- 作者机构：
- 作者简介：
- 基金信息：
  
  国家自然科学基金面上项目（81373535）;河南省科技攻关计划项目（142102310038）;河南省教育厅科学技术研究重点项目（14A360025）
- DOI：10.13422/j.cnki.syfjx.2016150134
  中图分类号： R735.1
- 纸质出版日期：2016
- 稿件说明：
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杨联河, 赵雪艳, 郭彬, 等. 启膈散与顺铂对食管癌细胞EC9706低氧下生长抑制及其miR-21,PDCD4和PTEN表达的影响[J]. 中国实验方剂学杂志, 2016,22(15):134-138.

YANG Lian-he, ZHAO Xue-yan, GUO Bin, et al. Inhibitive Effect of Qige San and Cisplatin on Growth of Esophageal Carcinoma Cells EC9706 and Its miR-21, PDCD4 and PTEN Expressions Under Hypoxia[J]. Chinese journal of experimental traditional medical formulae, 2016, 22(15): 134-138.
杨联河, 赵雪艳, 郭彬, 等. 启膈散与顺铂对食管癌细胞EC9706低氧下生长抑制及其miR-21,PDCD4和PTEN表达的影响[J]. 中国实验方剂学杂志, 2016,22(15):134-138. DOI： 10.13422/j.cnki.syfjx.2016150134.

YANG Lian-he, ZHAO Xue-yan, GUO Bin, et al. Inhibitive Effect of Qige San and Cisplatin on Growth of Esophageal Carcinoma Cells EC9706 and Its miR-21, PDCD4 and PTEN Expressions Under Hypoxia[J]. Chinese journal of experimental traditional medical formulae, 2016, 22(15): 134-138. DOI： 10.13422/j.cnki.syfjx.2016150134.

摘要

目的：观察低氧下启膈散和顺铂抑制食管癌细胞增殖的协同效应，从miR-21及其靶基因角度探讨其机制。方法：制备含药血清，噻唑蓝（MTT）比色法检测启膈散和顺铂单用或联合应用对食管癌细胞EC9706增殖的影响，实时荧光定量PCR（Real-time PCR）检测启膈散和顺铂单用或两者联合应用对食管癌细胞miR-21，程序死亡蛋白4（PDCD4），磷酸酶和张力蛋白类似物（PTEN）mRNA表达的影响，免疫印迹法（Western blot）检测PDCD4，PTEN蛋白表达。结果：启膈散在浓度为8%，10%与1 mg·L-1顺铂合用具有协同作用，其两药相互作用系数（CDI）分别为0.90，0.94。顺铂高浓度组，联合高、低浓度组miRNA-21表达量明显低于空白组（P<0.05）；联合高、低浓度组miRNA-21表达量显著低于同浓度顺铂组（P<0.01），且联合高浓度组低于联合低浓度组（P<0.05）。顺铂高浓度、启膈散组PDCD4 mRNA表达量明显低于空白组（P<0.05），而联合高、低浓度组PDCD4，PTEN mRNA表达量明显高于空白组（P<0.05）；联合高、低浓度组PDCD4，PTEN mRNA明显高于同浓度顺铂组（P<0.01），且联合高浓度组PTEN mRNA表达高于联合低浓度组（P<0.01）。与空白组比较，联合高、低浓度组PTEN蛋白表达明显升高（P<0.05），且联合高、低浓度组PTEN蛋白显著高于同浓度顺铂组（P<0.01）；顺铂高、低浓度组，联合高、低浓度组PDCD4明显高于空白组（P<0.05），联合高浓度组明显高于同浓度顺铂组和联合低浓度组（P<0.05，P<0.01）。结论：启膈散含药血清和顺铂在抑制食管癌细胞EC9706增殖方面具有协同作用，其机制可能与通过抑制miR-21表达从而升高PDCD4和PTEN基因表达相关。

Abstract

Objective: To investigate the synergistic effect of Qige San and cisplatin in inhibiting proliferation of esophageal carcinoma cells in hypoxia

in order to explore the mechanism from the perspective of miR-21 and its target genes. Method: Drug-containing serum was prepared. The single or combined effects of Qige San and cisplatin on cells activation were tested by 3-(4

5-dimethyl-2-thiazolyl)-2

5-diphenyl-2-H-tetrazolium bromid (MTT) assays. Apoptosis and cell cycle arrest induced by Qige San and cisplatin were detected by flow cytometry. MiR-21

programmed cell death 4 (PDCD4)

phosphatase and tensin homolog (PTEN) mRNA expressions were detected by Real-time PCR

while PDCD4 and PTEN protein expression were detected by Western blot. Result: Qige San in concentrations of 8%and 10%combined with 1 mg·L-1 cisplatin has a synergistic effect in inhibiting proliferation of esophageal carcinoma cells

with the coefficients of drug in interaction (CDI) of 0.90

0.94

respectively. The expressions of miRNA-21 in cisplatin high concentration group

and combination high and low concentration group were significantly lower than that in the control group(P<0.05). The expression of miRNA-21 in combination high and low concentration group was significantly lower than that of cisplatin groups with the same concentration (P<0.01)

and the low concentration group (P<0.05). The expression of PDCD4 mRNA was significantly lower in the cisplatin high concentration group and the Qige San group than in control group (P<0.05)

whereas the expression of PTEN and PDCD4 mRNA in combination high and low concentration group was significantly higher than that in control group (P<0.05)

and PDCD4

PTEN mRNA expressions in combination high and low concentration group was significantly higher than that of cisplatin groups with the same concentration (P<0.01)

and PTEN mRNA in high concentration combination group was higher than that in lower concentration combination group (P<0.01). Compared with the control group

the expression of PTEN protein was significantly higher in combination high and low concentration group(P<0.05)

and that in combination high and low concentration group was significantly higher than that in the same concentration of cisplatin group (P<0.01). PDCD4 was significantly higher in cisplatin high and low concentration group and combination high and low concentration group that in control group (P<0.05)

and combination high concentration group was significantly higher than that in cisplatin group and combination low concentration group (P<0.05

P<0.01). Conclusion: Qige San and cisplatin have a synergistic effect in inhibiting proliferation of esophageal carcinoma cell line EC9706

and the mechanism may be related to increase the expression of PDCD4 and PTEN by inhibiting miRNA-21 expression.

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