消痰解郁方对乳腺癌前病变MCF-10AT细胞PI3K/Akt通路的影响及机制
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第二军医大学附属长征医院中医科 上海,第二军医大学附属长征医院中医科 上海,第二军医大学附属长征医院中医科 上海,第二军医大学附属长征医院中医科 上海,第二军医大学附属长海医院中医科 上海

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上海市名老中医学术经验研究工作室建设项目(ZYSNXD-CC-MZY054),军队中医药科研专项基金(2010z131).


Effect of Xiaotan Jieyu Decoction on PI3K/Akt pathway of MCF-10AT cells and its mechanism
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Department of Traditional Chinese medicine,Changzheng Hospital Affiliated to the Second Military Medicine University,Department of Traditional Chinese medicine,Changzheng Hospital Affiliated to the Second Military Medicine University,Department of Traditional Chinese medicine,Changzheng Hospital Affiliated to the Second Military Medicine University,Department of Traditional Chinese medicine,Changzheng Hospital Affiliated to the Second Military Medicine University,Department of Traditional Chinese medicine,Changhai Hospital Affiliated to the Second Military Medicine University

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    摘要:

    目的 探讨消痰解郁方对乳腺癌前病变MCF-10AT细胞PI3K/Akt信号通路的影响及机制。方法 将MCF-10AT细胞分为消痰解郁方组、PI3K激酶选择性抑制剂LY294002组(抑制剂组)、消痰解郁方+抑制剂组和对照组。给予相应药物干预24、48 h后,采用CCK-8法观察各组细胞增殖及生长情况;药物干预48 h后,采用流式细胞仪检测各组细胞周期变化,用蛋白质印迹法检测各组细胞PTEN、PI3K、Akt蛋白表达变化。结果 药物干预24和48 h后,消痰解郁方组、抑制剂组和消痰解郁方+抑制剂组MCF-10AT细胞增殖受到抑制,且消痰解郁方+抑制剂组抑制率高于消痰解郁方组和抑制剂组,差异有统计学意义(P<0.05);药物干预48 h后,消痰解郁方组、抑制剂组和消痰解郁方+抑制剂组MCF-10AT细胞G0/G1期细胞比例增高,S期及G2/M期细胞比例下降,与对照组比较差异有统计学意义(P<0.05),其中消痰解郁方+抑制剂组与其他各组相比差异有统计学意义(P<0.05);药物干预48 h后,消痰解郁方组、抑制剂组、消痰解郁方+抑制剂组PI3K、p-Akt 蛋白表达较对照组减弱,PTEN蛋白表达较对照组增强,差异均有统计学意义(P<0.05)。结论 消痰解郁方可能通过抑制PI3K/AKT信号通路发挥其对MCF-10AT细胞的抑制及促凋亡作用。

    Abstract:

    Objective To study the effect of Xiaotan Jieyu (XTJY) Decoction on PI3K/Akt signal pathway of MCF-10AT cells of breast precancerous lesion and the ralated mechanism. Methods The MCF-10AT cells were randomly divided into following groups: XTJY group, PI3K/Akt inhibitor LY294002 (LY) group, XTJY+LY group and control (CON) group. After corresponding drug intervention, CCK-8 method was used to observe the MCF-10AT cell proliferation and growth inhibition at 24 h and 48 h, flow cytometry was used to detect the changes of cell cycle at 48 h, and the expression changes of PTEN, PI3K and Akt protein were detected by Western blotting analysis at 48 h. Results The proliferation of MCF-10AT cells were significantly inhibited at 24 h and 48 h in XTJY, LY and XTJY+LY groups, and the inhibitory rate of MCF-10AT cells in XTJY+LY group was significantly higher than those in XTJY and LY groups (P<0.05). Compared with CON group, the percentages of G0/G1 phase cells in XTJY, LY and XTJY+LY groups were significantly increased at 48 h (P<0.05), while the percentages of S phase and G2/M phase cells were significantly decreased (P<0.05); besides, there were significant differences between XTJY+LY group and the other two groups (P<0.05). Compared with CON group, the expressions of PI3K and p-AKT protein in XTJY, LY and XTJY+LY groups were significantly decreased at 48 h (P<0.05), and the expression of PTEN protein was significantly increased (P<0.05). Conclusion XTJY prescription may exert inhibitory and apoptotic effect on MCF-10AT cells through inhibiting the PI3K/Akt signal pathway.

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  • 收稿日期:2017-02-10
  • 最后修改日期:2017-03-13
  • 录用日期:2017-03-23
  • 在线发布日期: 2017-05-02
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