山奈酚诱导三阴性乳腺癌细胞MDA-MB-231中乳腺癌耐药蛋白的表达并下调抗肿瘤药物对MDA-MB-231细胞的杀伤作用
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摘要
为确定三叶青活性物质山奈酚对三阴性乳腺癌(triple negative breast cancer,TNBC)的影响及其作用的分子机制,通过定量PCR的方法检测TNBC临床标本及MDA-MB-231细胞中孕烷X受体(pregnane X receptor,PXR)的表达情况;通过荧光素酶报告基因活性检测确定山奈酚对MDA-MB-231细胞中PXR转录因子活性的影响;通过MTT(四唑盐)方法、裸鼠皮下成瘤方法研究了山奈酚以及抗肿瘤药物卡铂、维利帕尼以及拉帕替尼等对MDA-MB-231细胞的抗肿瘤作用。结果表明:PXR在TNBC临床标本以及MDA-MB-231细胞中有明确表达;山奈酚能够诱导MDA-MB-231细胞对抗肿瘤药物卡铂、维利帕尼以及拉帕替尼等对MDA-MB-231细胞的杀伤作用;山奈酚能够诱导MDA-MB-231中PXR的转录因子活性以及PXR下游基因乳腺癌的表达;使用小干扰RNA(small interfere RNA,siRNA)抑制乳腺癌耐药蛋白(breast cancer resistance protein,BCRP)的表达能够逆转山奈酚诱导的抗肿瘤药物耐药。可见,山奈酚诱导TNBC细胞MDA-MB-231中乳腺癌耐药蛋白的表达并下调抗肿瘤药物对MDA-MB-231细胞的杀伤作用。
In order to reveal the effect of Kaempferol on MDA-MB-231,a triple negative breast cancer (TNBC)cell line,the expression of pregnane X receptor (PXR) were examined in TNBC clinical specimens or cell line by qPCR experiments. Luciferase experiments was used to investigate the effect of Kaempferol inducing the transcription factor of PXR and qPCR experiments were used to examine the effect of Kaempferol inducing the expression of breast cancer resistance protein (BCRP) in MDA-MB-231 cells. Antitumor effect of agents, Carboplatin,Kaempferol,Veliparib or Lapatinib was examined by MTT assays. The effect of Kaempferol on MDA-MB-231 cells were further confirmed by subcutaneous tumor model in nude mice. The results show that the expression of PXR is examined in TNBC clinical specimens and MDA-MB-231 cells. Treatment of Kaempferol induces the transcription factor activation of PXR and induces the expression of BCRP in a PXR dependent manner. Results from MTT and subcutaneous tumor model indicate that treatment of Kaempferol induces the drug-resistance of MDA-MB-231 cells to antitumor agents in a BCRP dependent manner. It is concluded that Kaempferol induces on the expression of BCRP in TNBC cell line MDA-MB-231 and enhances the resistance of cells to antitumor agents.
In order to reveal the effect of Kaempferol on MDA-MB-231,a triple negative breast cancer (TNBC)cell line,the expression of pregnane X receptor (PXR) were examined in TNBC clinical specimens or cell line by qPCR experiments. Luciferase experiments was used to investigate the effect of Kaempferol inducing the transcription factor of PXR and qPCR experiments were used to examine the effect of Kaempferol inducing the expression of breast cancer resistance protein (BCRP) in MDA-MB-231 cells. Antitumor effect of agents, Carboplatin,Kaempferol,Veliparib or Lapatinib was examined by MTT assays. The effect of Kaempferol on MDA-MB-231 cells were further confirmed by subcutaneous tumor model in nude mice. The results show that the expression of PXR is examined in TNBC clinical specimens and MDA-MB-231 cells. Treatment of Kaempferol induces the transcription factor activation of PXR and induces the expression of BCRP in a PXR dependent manner. Results from MTT and subcutaneous tumor model indicate that treatment of Kaempferol induces the drug-resistance of MDA-MB-231 cells to antitumor agents in a BCRP dependent manner. It is concluded that Kaempferol induces on the expression of BCRP in TNBC cell line MDA-MB-231 and enhances the resistance of cells to antitumor agents.
引文
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18 Jia H,Yang Q,Wang T,et al.Rhamnetin induces sensitization of hepatocellular carcinoma cells to a small molecular kinase inhibitor or chemotherapeutic agents[J].Biochimica Biophysica Acta,2016,1860(7):1417-1430
2 Reynolds A,Mann J,Cummings J,et al.Carbohydrate quality and human health:a series of systematic reviews and meta-analyses[J].Lancet,2019,393(10170):434-445
3 Chen W,Zheng R,Baade P D,et al.Cancer statistics in China,2015[J].CA Cancer Journal for clinicians,2016,66(2):115-132
4 Wang J,Gan C,Retmana I A,et al.P-glycoprotein(MDR1/ABCB1)and breast cancer resistance Protein(BCRP/ABCG2)limit brain accumulation of the FLT3 inhibitor quizartinib in mice[J].International Journal of Phamaceutsics,2018,556:172-180
5 Jend6elovsky R,Jend6elovskáZ,KuchárováB,et al.Breast cancer resistance protein is the enemy of hypericin accumulation and toxicity of hypericin-mediated photodynamic therapy[J].Biomed Pharmacother,2019,109:2173-2181
6 Yang Q,Feng F,Zhang F,et al.LINE-1 ORF-1p functions as a novel HGF/ETS-1 signaling pathway co-activator and promotes the growth of MDA-MB-231 cell[J].Cell Signal,2013,25(12):2652-2660
7 Feng F,Jiang Q,Cao S,et al.Pregnane X receptor mediates sorafenib resistance in advanced hepatocellular carcinoma[J].Biochimica Biophysica Acta,2018,1862(4):1017-1030
8 Feng F,Lu Y Y,Zhang F,et al.Long interspersed nuclear element ORF-1 protein promotes proliferation and resistance to chemotherapy in hepatocellular carcinoma[J].World Journal of Gastroenterol,2013,19(7):1068-1078
9 Rady M,Mostageer M,Rohde J,et al.Therapy-relevant aberrant expression of MRP3 and BCRP mRNA in TCC-/SCC-bladder cancer tissue of untreated patients[J].Oncology Reports,2017,38(1):551-560
10 An L,Li D D,Chu H X,et al.Terfenadine combined with epirubicin impedes the chemo-resistant human non-small cell lung cancer both in vitro and in vivo through EMT and Notch reversal[J].Pharmacological Research,2017,124:105-115
11 Shao Z,Li Y,Dai W,et al.ETS-1 induces Sorafenib-resistance in hepatocellular carcinoma cells via regulating transcription factor activity of PXR[J].Pharmacological Research,2018,135:188-200
12 Cui J,Germer K,Wu T,et al.Cross-talk between HER2 and MED1 regulates tamoxifen resistance of human breast cancer cells[J].Cancer Research,2012,72(21):5625-5634
13 Sang M,Meng L,Ma C,et al.Effect of AR antagonist combined with PARP1 inhibitor on sporadic triple-negative breast cancer bearing AR expression and methylation-mediated BRCA1dysfunction[J].Biomed Pharmacother,2019,111:169-177
14 Wang Z,Wang N,Li W,et al.Caveolin-1 mediates chemoresistance in breast cancer stem cells viaβ-catenin/ABCG2 signaling pathway[J].Carcinogenesis,2014,35(10):2346-2356
15 Mackowiak B,Hodge J,Stern S,et al.The roles of xenobiotic receptors:beyond chemical disposition[J].Drug Metabolism&Disposition,2018,46(9):1361-1371
16 Campbell S D,Gadel S,Friedel C,et al.Influence of HIV antiretrovirals on methadone N-demethylation and transport[J].Biochemical Pharmacology,2015,95(2):115-125
17 Qiao E Q,Yang H J.Effect of pregnane X receptor expression on drug resistance in breast cancer[J].Oncology Letters,2014,7(4):1191-1196
18 Jia H,Yang Q,Wang T,et al.Rhamnetin induces sensitization of hepatocellular carcinoma cells to a small molecular kinase inhibitor or chemotherapeutic agents[J].Biochimica Biophysica Acta,2016,1860(7):1417-1430
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