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山东中医药大学 第一临床医学院,中医学院,济南 250014
王楠,硕士,从事中医外科学乳腺甲状腺研究,E-mail: 1156004128@qq.com
刘晓菲,副教授,副主任医师,从事乳腺甲状腺外科的中医及中西医结合临床、教学及科研工作,E-mail: drliuxf@126.com
收稿日期:2018-10-17,
网络出版日期:2018-11-05,
纸质出版日期:2019-03-05
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王楠, 刘晓菲, 张洋, 等. 阳和化岩汤对HER-2高表达型乳腺癌细胞PI3K/Akt通路HER-2,PI3K,p-Akt表达的机制[J]. 中国实验方剂学杂志, 2019,25(5):98-104.
Nan WANG, Xiao-fei LIU, Yang ZHANG, et al. Effect of Yanghe Huayan Tang Combined with PI3K Inhibitor on Expressions of HER-2, PI3K and p-Akt in PI3K/Akt Pathway of High-expression HER-2 Breast Cancer Cell Lines[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(5): 98-104.
王楠, 刘晓菲, 张洋, 等. 阳和化岩汤对HER-2高表达型乳腺癌细胞PI3K/Akt通路HER-2,PI3K,p-Akt表达的机制[J]. 中国实验方剂学杂志, 2019,25(5):98-104. DOI: 10.13422/j.cnki.syfjx.20190425.
Nan WANG, Xiao-fei LIU, Yang ZHANG, et al. Effect of Yanghe Huayan Tang Combined with PI3K Inhibitor on Expressions of HER-2, PI3K and p-Akt in PI3K/Akt Pathway of High-expression HER-2 Breast Cancer Cell Lines[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(5): 98-104. DOI: 10.13422/j.cnki.syfjx.20190425.
目的:
2
观察阳和化岩汤与磷脂酰肌醇-3羟基激酶(PI3K)抑制剂LY294002对乳腺癌细胞株SK-BR-3[人表皮生长因子受体-2(HER-2)高表达型]PI3K/蛋白激酶B(Akt)信号通路中HER-2,PI3K,磷酸化蛋白激酶B(p-Akt)表达的影响,研究阳和化岩汤的干预机制。
方法:
2
制备阳和化岩汤肠吸收液,将乳腺癌细胞株SK-BR-3分为空白组,阳和化岩汤组,LY294002组,LY294002+阳和化岩汤组,阳和化岩汤(125g·L
-1
),LY294002(0.05g·L
-1
),阳和化岩汤+LY294002(阳和化岩汤125g·L
-1
,LY294002 0.05g·L
-1
),用药干预24h后通过免疫组化、蛋白免疫印迹法(Western blot)检测HER-2,PI3K,p-Akt蛋白表达,通过逆转录聚合酶链式反应(RT-PCR)检测HER-2,PI3K,p-AktmRNA表达。
结果:
2
与空白组比较,LY294002组,LY294002+阳和化岩汤组均能抑制乳腺癌细胞株HER-2,PI3K,p-Akt蛋白及mRNA表达(
P
<
0.05);阳和化岩汤组能抑制乳腺癌细胞株HER-2,p-Akt蛋白及mRNA表达(
P
<
0.05),但对PI3K的抑制作用不明显;联用LY294002较单用LY294002具有进一步抑制PI3K表达的作用(
P
<
0.05)。与阳和化岩汤组,LY294002组比较,LY294002+阳和化岩汤组能够明显抑制HER-2,PI3K,p-Akt蛋白及mRNA表达(
P
<
0.05)。
结论:
2
阳和化岩汤能够抑制乳腺癌脉管生成和侵袭,主要作用机制为通过干预PI3K/Akt通路,降低通路中HER-2,PI3K,p-Akt表达。
Objective:
2
To investigate effect of Yanghe Huayan Tang and phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 in intervening expressions of human epidermal growth factor receptor human epidermal growth factor receptor-2(HER-2)
PI3K and phosphorylated serine/threonine kinase (p-Akt)in PI3K/Akt pathway of breast cancer cell line SK-BR-3 (HER-2 high expression)
in vitro
and study intervention mechanism of Yanghe Huayan Tang.
Method:
2
Yanghe Huayan Tang intestinal absorption fluid was prepared
breast cancer cell strain SK-BR-3 was divided into blank group
Yanghe Huayan Tang group
LY294002 group
LY294002+ Yanghe Huayan Tang group. The concentration was respectively 125 g·L
-1
for Yanghe Huayan Tang
0.05 g·L
-1
for LY294002
125 g·L
-1
for Yanghe Huayan Tang+ LY294002.The expressions of HER-2
PI3K and p-Akt protein were detected by immunohistochemistry and Western blot after 24 h. The most appropriate and effective concentration was obtained through extensive experiments. The expressions of HER-2
PI3K and p-Akt were detected by reverse transcription polymerase chain reaction (RT-PCR).
Result:
2
Compared with blank group
LY294002 group
and LY294002+ Yanghe Huayan Tang group could inhibit protein and mRNA expressions of HER-2
PI3K
p-Akt in breast cancer cell lines(
P
<
0.05). Yanghe Huayan Tang group could inhibit expressions of HER-2
p-Akt and mRNA (
P
<
0.05) in breast cancer cell lines
but inhibitory effect on PI3K was not obvious; combination with LY294002 could further inhibit PI3K expression compared with single administration with LY294002 (
P
<
0.05). Compared with LY294002 group
LY294002+ Yanghe Huayan Tang group had a more obvious inhibitory effect. The combined administration of Yanghe Huayan Tang and LY294002 could inhibit protein and mRNA expressions of HER-2
PI3K
p-Akt significantly (
P
<
0.05).
Conclusion:
2
Yanghe Huayan Tang can inhibit angiogenesis and invasion of breast cancer. Its main mechanism is expressed by intervening PI3K/Akt pathway
reducing expressions of HER-2
PI3K and p-Akt in pathway.
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