[1]魏昌晟,沈义军,张智,等.苦参碱逆转乳腺癌耐药株MCF-7/ADR多药耐药与PI3K/AKT通道的关系[J].第三军医大学学报,2014,36(22):2254-2258.
 Wei Changsheng,Shen Yijun,Zhang Zhi,et al.Matrine reverses multidrug resistance in breast cancer drug-resistant cell line MCF-7/ADR through inhibiting PI3K/AKT signal pathway[J].J Third Mil Med Univ,2014,36(22):2254-2258.
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苦参碱逆转乳腺癌耐药株MCF-7/ADR多药耐药与PI3K/AKT通道的关系(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
36卷
期数:
2014年第22期
页码:
2254-2258
栏目:
论著
出版日期:
2014-11-30

文章信息/Info

Title:
Matrine reverses multidrug resistance in breast cancer drug-resistant cell line MCF-7/ADR through inhibiting PI3K/AKT signal pathway
作者:
魏昌晟沈义军张智王健周炳刚
宁夏医科大学: 研究生院,宁夏颅脑疾病重点实验室;宁夏回族自治区人民医院乳腺微创外科;银川市第二人民医院普外科
Author(s):
Wei Changsheng Shen Yijun Zhang Zhi Wang Jian Zhou Binggang

School of Postgraduate, Ningxia Key Laboratory for Cerebrocranial Diseases, Ningxia Medical University, Yinchuan, Ningxia Hui Autonomous Region, 750004; Department of Breast Minimally Invasive Surgery, People’s Hospital of Ningxia Hui Autonomous Region, Yinchuan, Ningxia Hui Autonomous Region, 750002; Department of General Surgery, the Second People’s Hospital of Yinchuan, Yinchuan, Ningxia Hui Autonomous Region, 750021, China

关键词:
苦参碱 乳腺癌耐药株MCF-7/ADR 多药耐药 PI3K/AKT信号通路
Keywords:
matrine MCF-7/ADR multidrug resistance PI3K/AKT signal pathway
分类号:
R285.5;R73-362;R737.9
文献标志码:
A
摘要:

目的      探索苦参碱对乳腺癌耐药株MCF-7/ADR多药耐药的逆转作用及其机制。      方法      采用MTT法检测不同浓度苦参碱和MK2206作用MCF-7/ADR细胞24 h后的生长抑制率,以抑制率为10%的药物浓度为检测标准;用荧光RT-PCR、Western blot法检测不同浓度苦参碱作用MCF-7/ADR细胞24 h后MDR1、MRP1、PTEN、AKT基因mRNA和蛋白的表达。      结果      荧光定量RT-PCR结果显示0.6、1.2 g/L的苦参碱干预组与空白对照组相比,随着苦参碱浓度的提高,耐药基因MDR1、MRP1的表达量逐渐降低(0.6 g/L苦参碱组MDR1基因的相对表达量为0.659±0.074,MRP1基因的相对表达量为0.503±0.058,与空白对照组相比P<0.01);PI3K/AKT信号通路中的PTEN基因表达量逐渐增高而AKT基因的表达量逐渐降低;相同抑制率的苦参碱组(0.6 g/L)和MK2206组(0.05 μmol/L)相比,两组降低MDR1、MRP1、AKT基因表达量的差异不明显(P>0.05),MK2206组更能增高PTEN 基因的表达量。Western blot检测结果显示0.3、0.6、1.2 g/L 不同浓度苦参碱干预组与空白对照组相比,随着苦参碱浓度的提高耐药蛋白p-gp、MRP1表达量逐渐降低(0.6 g/L苦参碱组p-gp蛋白的相对表达量为0.316±0.033,MRP1蛋白的相对表达量为0.134±0.014,与空白对照组相比P<0.01),PI3K/AKT信号通路中的p-AKT蛋白表达量逐渐降低,PTEN蛋白表达量逐渐增高,总AKT基因表达量变化不明显;相同抑制率的苦参碱组(0.6 g/L)和MK2206组(0.05 μmol/L)相比,苦参碱组更能降低p-gp、MRP1的蛋白表达量且更能增高PTEN的蛋白表达量,但MK2206组更能降低p-AKT的蛋白表达量,对总AKT蛋白表达量两组差异不明显(P>0.05)。      结论      苦参碱具有逆转乳腺癌多药耐药的作用,其机制可能是通过抑制PI3K/AKT通道发挥作用。

Abstract:

Objective      To explore the bioactivities of matrine and PI3K/AKT signal pathway in the reversal of multidrug resistance in breast cancer cells.       Methods      MTT assay was used to measure the inhibitory rate of matrine of different concentrations on breast cancer drug-resistant cell line MCF-7/ADR after 24 h. In addition, real-time RT-PCR and Western blotting were used to evaluate the mRNA and protein expression levels of MDR1, MRP1, PTEN, AKT, and p-AKT in MCF-7/ADR cells.       Results      Real-time RT-PCR results showed when compared with the control group, the relative expression levels of MDR1 and MRP1 in the 0.6 and 1.2 g/L matrine groups were reduced gradually (MDR1 0.659±0.074 and MRP1 0.503±0.058 in the 0.6 g/L matrine group vs MDR1 0.438±0.034 and MRP1 0.399±0.025 in the 1.2 g/L matrine group, P<0.01).The relative gene expression level of AKT was also reduced gradually, but that of PTEN was increased gradually. The expression levels of MDR1, MRP1, and AKT in the cells intervened separately by 0.6 g/L matrine and 0.05 μmol/L, MK2206 didn’t have significant difference at the same inhibition rate. Western blotting results showed when compared with the control group, the expression levels of p-gp and MRP1 of the 0.3, 0.6 and 1.2 g/L matrine groups were reduced gradually (p-gp 0.316±0.033 and MRP1 0.134±0.014 in the 0.6 g/L matrine group, P<0.01).The protein expression of p-AKT was also reduced gradually, but that of PTEN was increased gradually. The total AKT protein expression didn’t change obviously (P>0.05). At the same inhibitory rate of matrine (0.6 g/L) and MK2206 (0.05 μmol/L), the matrine group was more effective than MK2206 group in reducing the protein expression of p-gp, MRP1 and p-AKT and increasing the protein expression of PTEN. However, the total AKT protein expression of the 2 groups didn’t have significant difference (P>0.05).       Conclusion      The effect of matrine mediating the reversal of drug resistance of breast cancer cells probably involves PI3K/AKT signal pathway.

参考文献/References:

[1]Ferlay J, Shin H R, Bray F, et al. Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008[J]. Int J Cancer, 2010, 127(12): 2893-2917. 
[2]Li T, Wong V K, Yi X Q, et al. Matrine induces cell anergy in human Jurkat T cells through modulation of mitogen-activated protein kinases and nuclear factor of activated T-cells signaling with concomitant up-regulation of anergy-associated genes expression[J]. Biol Pharm Bull, 2010, 33(1): 40-46. 
[3]Cheng H, Xia B, Zhang L, et al. Matrine improves 2, 4, 6-trinitrobenzene sulfonic acid-induced colitis in mice[J]. Pharmacol Res, 2006, 53(3): 202-208. 
[4] Li X, Chu W, Liu J, et al. Antiarrhythmic properties of long-term treatment with matrine in arrhythmic rat induced by coronary ligation[J]. Biol Pharm Bull, 2009, 32(9): 1521-1526.
[5] Dai Z J, Gao J, Ji Z Z, et al. Matrine induces apoptosis in gastric carcinoma cells via alteration of Fas/FasL and activation of caspase-3[J]. J Ethnopharmacol, 2009, 123(1): 91-96. 
[6]Zhang J, Li Y, Chen X, et al. Autophagy is involved in anticancer effects of matrine on SGC-7901 human gastric cancer cells[J]. Oncol Rep, 2011, 26(1): 115-124. 
[7]Zhang Y, Zhang H, Yu P, et al. Effects of matrine against the growth of human lung cancer and hepatoma cells as well as lung cancer cell migration[J]. Cytotechnology, 2009, 59(3): 191-200. 
[8]Zhang J Q, Li Y M, Liu T, et al. Antitumor effect of matrine in human hepatoma G2 cells by inducing apoptosis and autophagy[J]. World J Gastroenterol, 2010, 16(34): 4281-4290.
[9]Yu P, Liu Q, Liu K, et al. Matrine suppresses breast cancer cell proliferation and invasion via VEGF-Akt-NF-kappaB signaling[J]. Cytotechnology, 2009, 59(3): 219-229.
[10]Han Y, Zhang S, Wu J, et al. Matrine induces apoptosis of human multiple myeloma cells via activation of the mitochondrial pathway[J]. Leuk Lymphoma, 2010, 51(7): 1337-1346. 
[11]孙付军, 王宁, 李贵海, 等. 苦参碱对获得性多药耐药小鼠S180肿瘤细胞基因表达产物P170、LRP及TOPOⅡ表达的影响[J]. 中药材, 2004, 27(11): 838-840. 
[12]李贵海, 王玫, 孙付军, 等. 苦参碱逆转小鼠S180肿瘤细胞获得性多药耐药基因相关表达产物过度表达的研究[J]. 中药材, 2006, 29(1): 40-42. 
[13]Luo S X, Deng W Y, Wang X F, et al. Molecular mechanism of indirubin-3′-monoxime and Matrine in the reversal of paclitaxel resistance in NCI-H520/TAX25 cell line[J]. Chin Med J (Engl), 2013, 126(5): 925-929. 
[14]Lindsley C W. The Akt/PKB family of protein kinases: a review of small molecule inhibitors and progress towards target validation: a 2009 update[J]. Curr Top Med Chem, 2010, 10(4): 458-477. 
[15]He S, Liu F, Xie Z, et al. P-Glycoprotein/MDR1 regulates pokemon gene transcription through p53 expression in human breast cancer cells[J]. Int J Mol Sci, 2010, 11(9): 3039-3051. 
[16]Kim D, Cheng G Z, Lindsley C W, et al. Targeting the phosphatidylinositol-3 kinase/Akt pathway for the treatment of cancer[J]. Curr Opin Investig Drugs, 2005, 6(12): 1250-1258. [17]Tian T, Nan K J, Guo H, et al. PTEN inhibits the migration and invasion of HepG2 cells by coordinately decreasing MMP expression via the PI3K/Akt pathway[J]. Oncol Rep, 2010, 23(6): 1593-1600.
[18]Lee C S, Kim Y J, Jang E R, et al. Akt inhibitor enhances apoptotic effect of carboplatin on human epithelial ovarian carcinoma cell lines[J]. Eur J Pharmacol, 2010, 632(1/3): 7-13. 
[19]Burris H A 3rd. Overcoming acquired resistance to anticancer therapy: focus on the PI3K/AKT/mTOR pathway[J]. Cancer Chemother Pharmacol, 2013, 71(4): 829-842. 
[20]Calderaro J, Rebouissou S, de-Koning L, et al. PI3K/AKT pathway activation in bladder carcinogenesis[J]. Int J Cancer, 2014, 134(8): 1776-1784.
[21] Tapia O, Riquelme I, Leal P, et al. The PI3K/AKT/mTOR pathway is activated in gastric cancer with potential prognostic and predictive significance[J]. Virchows Arch, 2014, 465(1): 25-33. 
[22]郭崇勇, 柯卫锋, 宋科瑛, 等. PI3K/AKT通路参与调控乳腺癌多药耐药和侵袭转移的研究[J]. 现代生物医学进展, 2012, 12(25): 4809-4812. 
[23]Liu F, Liu S, He S, et al. Survivin transcription is associated with P-glycoprotein/MDR1 overexpression in the multidrug resistance of MCF-7 breast cancer cells[J]. Oncol Rep, 2010, 23(5): 1469-1475. 
[24]Ren D, Jia L, Li Y, et al. ST6GalNAcII mediates the invasive properties of breast carcinoma through PI3K/Akt/NF-κB signaling pathway[J]. IUBMB Life, 2014, 66(4): 300-308. 
[25] Jeong Y J, Choi Y, Shin J M, et al. Melittin suppresses EGF-induced cell motility and invasion by inhibiting PI3K/Akt/mTOR signaling pathway in breast cancer cells[J]. Food Chem Toxicol, 2014, 68: 218-225. 
[26] Huemer F, Bartsch R, Gnant M. The PI3K/AKT/MTOR Signaling Pathway: The Role of PI3K and AKT Inhibitors in Breast Cancer[J]. Curr Breast Cancer Rep, 2014, 6(2): 59-70.

更新日期/Last Update: 2014-11-18