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
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摘要
目的: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: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(125 g·L-1),LY294002 (0. 05 g·L-1),阳和化岩汤+LY294002 (阳和化岩汤125 g·L-1,LY294002 0. 05 g·L-1),用药干预24 h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-Akt mRNA表达。结果: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: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: To investigate effect of Yanghe Huayan Tang and phosphatidylinositol 3-kinase(PI3 K) inhibitor LY294002 in intervening expressions of human epidermal growth factor receptor human epidermal growth factor receptor-2(HER-2),PI3 K and phosphorylated serine/threonine kinase(p-Akt) in PI3 K/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: 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,PI3 K 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,PI3 K and p-Akt were detected by reverse transcription polymerase chain reaction(RT-PCR). Result: Compared with blank group,LY294002 group,and LY294002 + Yanghe Huayan Tang group could inhibit protein and mRNA expressions of HER-2,PI3 K, 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 PI3 K was not obvious; combination with LY294002 could further inhibit PI3 K 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,PI3 K,p-Akt significantly(P < 0. 05). Conclusion: Yanghe Huayan Tang can inhibit angiogenesis and invasion of breast cancer. Its main mechanism is expressed by intervening PI3 K/Akt pathway,reducing expressions of HER-2,PI3 K and p-Akt in pathway.
Objective: To investigate effect of Yanghe Huayan Tang and phosphatidylinositol 3-kinase(PI3 K) inhibitor LY294002 in intervening expressions of human epidermal growth factor receptor human epidermal growth factor receptor-2(HER-2),PI3 K and phosphorylated serine/threonine kinase(p-Akt) in PI3 K/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: 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,PI3 K 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,PI3 K and p-Akt were detected by reverse transcription polymerase chain reaction(RT-PCR). Result: Compared with blank group,LY294002 group,and LY294002 + Yanghe Huayan Tang group could inhibit protein and mRNA expressions of HER-2,PI3 K, 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 PI3 K was not obvious; combination with LY294002 could further inhibit PI3 K 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,PI3 K,p-Akt significantly(P < 0. 05). Conclusion: Yanghe Huayan Tang can inhibit angiogenesis and invasion of breast cancer. Its main mechanism is expressed by intervening PI3 K/Akt pathway,reducing expressions of HER-2,PI3 K and p-Akt in pathway.
引文
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[11]刘晓菲,李静蔚,李湘奇,等.阳和化岩汤对HER-2高表达型裸鼠荷瘤模型肿瘤血管生成的影响及机制的研究[J].中药材,2018,41(6):1468-1471.
[12]黄斌,李耿,郭宇飞,等.脑心通胶囊中4个成分肠吸收研究[J].中国中药杂志,2013,38(6):889-893.
[13]周阿高,李琰.中医药对乳腺癌患者生存质量影响的Meta分析[J].中国实验方剂学杂志,2012,18(5):220-222.
[14]鲍悦,高久堂,孙佳明,等.中药鹿角胶的研究进展[J].吉林中医药,2016,36(2):173-175,204.
[15]梁颖,徐绍娜,徐放,等.熟地黄多糖对环磷酰胺诱导小鼠的抗突变作用研究[J].中医药信息,2010,27(4):110-112.
[16]陈洋洋,王瑞莉,王斌,等.土贝母皂苷乙诱导非小细胞肺癌A549细胞凋亡的研究[J].中国中医急症,2017,26(8):1336-1339.
[17]袁理春,陈保生,吕丽芬.丽江山慈菇栽培技术研究初报[J].中国中药杂志,2003,28(6):98-99.
[18]赵志梅,张立杰,夏天,等.莪术主要单体成分抗炎、抗肿瘤作用研究进展[J].药物评价研究,2017,40(1):119-124.
[19]陈晓鹏,李昂,褚扬,等.养血清脑颗粒中有效成分的肠外翻吸收研究[J].中国实验方剂学杂志,2013,19(4):159-162.
[20] Nuciforo P,Thyparambil S,Aura C,et al. High HER-2protein levels correlate with increased survival in breast cancer patients treated with anti-HER-2 therapy[J].Mol Oncol,2016,10(1):138-147.
[21] Nitta H,Kelly B D,Allred C,et al. The assessment of HER-2 status in breast cancer:the past,the present,and the future[J]. Pathol Int,2016,66(6):313-324.
[22] Garcia-Garcia C,Ibrahim Y H,Serra V,et al. Dual m TORC1/2 and HER-2 blockade results in antitumor activity in preclinical models of breast cancer resistant to anti-HER-2 therapy[J]. Clin Cancer Res,2012,18(9):2603-2612.
[23] HU L,Zaloudek C,Mills G B,et al. In vivo and in vitro ovariancarcinoma growth inhibition by a phosphatidylinositol 3 kinase in-hibitor(LY294002)[J]. Clin Cancer Res,2009,6(3):880-886.
[2]胡升芳,谷焕鹏,陈红风,等.乳宁Ⅱ号方对HER-2过表达型乳腺癌PI3K/Akt信号通路的作用机制研究[J].中华中医药学刊,2017,35(4):980-983.
[3]曾佳佳,杨润祥,刘蓉.乳腺癌的靶向治疗研究进展[J].中国生化药物杂志,2016,36(1):7-11.
[4] Garcla C,lbrahim Y H,Serra V,et al. Dual m TORC1/2and HER-2 blockade results in antitumor activity in preclinical models of breast cancer resistant to antiHER-2 theragy[J]. Clin Cancer Res,2012,18(9):2603-2612.
[5]张刚,李中,林晓萌,等. PI3K/Akt信号通路在乳腺癌中的表达及其意义[J].中国妇幼保健,2015,30(20):3475-3478.
[6]肖洋炯,张萍,黄明辉,等.两种常用PI3K抑制剂对人乳腺癌耐药细胞株MCF-7/MIT耐药性的不同影响[J].肿瘤,2009,29(7):620-625.
[7]卞磊,陈光伟,马阳.温阳法治疗恶性肿瘤体会[J].实用中医药杂志,2012,28(10):872-873.
[8]刘晓菲,殷玉琨,王蕾,等.阳和化岩汤对大鼠乳腺癌癌前病变的抑制作用研究[J].中华中医药学刊,2014,32(2):284-287.
[9]李静蔚,刘晓菲,陈宏志.温阳散结法干预乳腺癌癌前病变MCF-10AT细胞的生长抑制及诱导凋亡研究[J].中医药学报,2013,41(4):41-45.
[10]李静蔚,刘晓菲,梁栋,等.阳和化岩汤对大鼠乳腺癌癌前病变细胞超微结构的影响[J].中医药学报,2014,42(1):42-46.
[11]刘晓菲,李静蔚,李湘奇,等.阳和化岩汤对HER-2高表达型裸鼠荷瘤模型肿瘤血管生成的影响及机制的研究[J].中药材,2018,41(6):1468-1471.
[12]黄斌,李耿,郭宇飞,等.脑心通胶囊中4个成分肠吸收研究[J].中国中药杂志,2013,38(6):889-893.
[13]周阿高,李琰.中医药对乳腺癌患者生存质量影响的Meta分析[J].中国实验方剂学杂志,2012,18(5):220-222.
[14]鲍悦,高久堂,孙佳明,等.中药鹿角胶的研究进展[J].吉林中医药,2016,36(2):173-175,204.
[15]梁颖,徐绍娜,徐放,等.熟地黄多糖对环磷酰胺诱导小鼠的抗突变作用研究[J].中医药信息,2010,27(4):110-112.
[16]陈洋洋,王瑞莉,王斌,等.土贝母皂苷乙诱导非小细胞肺癌A549细胞凋亡的研究[J].中国中医急症,2017,26(8):1336-1339.
[17]袁理春,陈保生,吕丽芬.丽江山慈菇栽培技术研究初报[J].中国中药杂志,2003,28(6):98-99.
[18]赵志梅,张立杰,夏天,等.莪术主要单体成分抗炎、抗肿瘤作用研究进展[J].药物评价研究,2017,40(1):119-124.
[19]陈晓鹏,李昂,褚扬,等.养血清脑颗粒中有效成分的肠外翻吸收研究[J].中国实验方剂学杂志,2013,19(4):159-162.
[20] Nuciforo P,Thyparambil S,Aura C,et al. High HER-2protein levels correlate with increased survival in breast cancer patients treated with anti-HER-2 therapy[J].Mol Oncol,2016,10(1):138-147.
[21] Nitta H,Kelly B D,Allred C,et al. The assessment of HER-2 status in breast cancer:the past,the present,and the future[J]. Pathol Int,2016,66(6):313-324.
[22] Garcia-Garcia C,Ibrahim Y H,Serra V,et al. Dual m TORC1/2 and HER-2 blockade results in antitumor activity in preclinical models of breast cancer resistant to anti-HER-2 therapy[J]. Clin Cancer Res,2012,18(9):2603-2612.
[23] HU L,Zaloudek C,Mills G B,et al. In vivo and in vitro ovariancarcinoma growth inhibition by a phosphatidylinositol 3 kinase in-hibitor(LY294002)[J]. Clin Cancer Res,2009,6(3):880-886.
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