β-elemene通过调节耐药特异性miRNA逆转乳腺癌细胞耐药性的机制研究
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摘要
目的:探讨β-榄香烯逆转乳腺癌细胞耐药性的机制,明确其抗肿瘤作用。方法:通过向MCF-7细胞株培养基内逐渐增加临床上最常用于乳腺癌化学治疗的药物多西紫杉醇(Doc)或阿霉素(Adr),逐渐提高培养基中药物浓度的方法成功地构建出亲本细胞株MCF-7中耐药亚型,选择乳腺癌阿霉素耐药细胞(MCF-7/Adr)和乳腺癌多西紫杉醇(MCF-7/Doc)耐药细胞株,采用MTT-cytotoxic、miRNA微阵列、实时定量PCR、双荧光素酶活性测定等方法,研究β-elemene对乳腺癌耐药细胞生存率、耐药特异性miRNA29a、miRNA452、miRNA34a及miRNA222表达的影响。结果:β-elemene明显降低耐药乳腺癌细胞株细胞生存率和增殖率,β-elemene显著调节MCF-7/Adr和MCF-7/Doc细胞内miRNAs的表达水平,其中6个miRNAs上调,12个miRNAs下调显著。结论:β-elemene是一种从植物中提取的天然抗肿瘤中药成分,可以逆转乳腺癌细胞的耐药性。证实了中药β-elemene可以通过调控耐药相关miRNA表达的机制抗乳腺癌耐药细胞的耐药性。
Objective: To explore the mechanism of the reversal of drug resistance in breast cancer cells and its antitumor effect. Methods: Through the MCF-7 cell lines within the medium increased the most commonly used clinical breast cancer chemotherapy drug Docetaxel(Doc) and Adriamycin(Adr), and gradually improve the drug concentrations in the culture medium method successfully build a parent cell line MCF-7 resistant sub-types, choose Adriamycin resistance of breast cancer cells(MCF-7/Adr) and breast cancer Docetaxel(MCF-7/Doc) drug resistant cell lines, the determined by MTT-cytotoxic, microRNAs microarray and real-time quantitative PCR, double luciferase activity determination methods, such as to investigate the effects of β-elemene on survival rate of breast cancer drug-resistant cells, drug-resistant specific miRNA-29 a, miRNA-452, miRNA-34 a and miRNA-222 expression. Results: The survival rate and proliferation rate of drug-resistant breast cancer cell lines were significantly reduced by β-elemene,while the expression level of miRNAs in McF-7/Adr and McF-7/Doc cells was significantly regulated by β-elemene,6 miRNAs of which were up-regulated and 12 of which were significantly down-regulated. Conclusion: β-elemene is a kind of natural anti-tumor Chinese medicine component extracted from plants, which can reverse the drug resistance of breast cancer cells. It was confirmed that β-elemene of traditional Chinese medicine could regulate the resistanceof breast cancer resistant cells by regulating the expression of drug-resist ant miRNA.
Objective: To explore the mechanism of the reversal of drug resistance in breast cancer cells and its antitumor effect. Methods: Through the MCF-7 cell lines within the medium increased the most commonly used clinical breast cancer chemotherapy drug Docetaxel(Doc) and Adriamycin(Adr), and gradually improve the drug concentrations in the culture medium method successfully build a parent cell line MCF-7 resistant sub-types, choose Adriamycin resistance of breast cancer cells(MCF-7/Adr) and breast cancer Docetaxel(MCF-7/Doc) drug resistant cell lines, the determined by MTT-cytotoxic, microRNAs microarray and real-time quantitative PCR, double luciferase activity determination methods, such as to investigate the effects of β-elemene on survival rate of breast cancer drug-resistant cells, drug-resistant specific miRNA-29 a, miRNA-452, miRNA-34 a and miRNA-222 expression. Results: The survival rate and proliferation rate of drug-resistant breast cancer cell lines were significantly reduced by β-elemene,while the expression level of miRNAs in McF-7/Adr and McF-7/Doc cells was significantly regulated by β-elemene,6 miRNAs of which were up-regulated and 12 of which were significantly down-regulated. Conclusion: β-elemene is a kind of natural anti-tumor Chinese medicine component extracted from plants, which can reverse the drug resistance of breast cancer cells. It was confirmed that β-elemene of traditional Chinese medicine could regulate the resistanceof breast cancer resistant cells by regulating the expression of drug-resist ant miRNA.
引文
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[2]林潇,胡倩,姚和瑞,等.转移性乳腺癌的化疗策略[J/CD].中华临床医师杂志电子版,2015,9(6):966-971.
[3]邓恒,束樱子,张珺.β-榄香烯在抗耐药机制中的研究进展及临床展望[J].辽宁中医杂志,2017,44(7):1558-1560.
[4]Guo HQ,Zhang GN,Wang YJ,et al.β-elemene,a compound derived from Rhizoma zedoariae,reverses multidrug resistance mediated by the ABCB1 transporter[J].Oncol Rep,2014,31(2):858-866.
[5]Li L,Yuan L,Luo J,et al.MiR-34a inhibits proliferation and migration of breast cancer through down-regulation of Bcl-2 and SIRT1[J].Clin Expmed,2013,13(2):109-117.
[6]齐创,向东华,廖天志.乳腺癌分子分型对多西他赛联合表阿霉素新辅助化疗疗效和预后的预测价值[J].中国肿瘤临床与康复,2016,23:17-21.
[7]Chen W,Zhang S,Zou X.Evaluation on the incidence,mortality and tendency of lung cancer in china[J].Thoracic Cancer,2010,1(1):35-40.
[8]Chen W,Zheng R,Zeng H,et al.Annual report on status of cancer in china,2011[J].中国癌症研究(英文版),2015,27(1):2-12.
[9]张斌,巩鹏,付虹,等.榄香烯在老年复发、转移性乳腺癌铂类联合化疗中的应用[J/CD].中华乳腺病杂志(电子版),2018,12(2):73-78.
[10]张斌.β-榄香烯与吉非替尼逆转乳腺癌MCF-7细胞他莫西芬耐药的实验研究[D].大连:大连医科大学,2012.
[11]刘慧,芮景.局部晚期乳腺癌新辅助化疗[J].国际肿瘤学杂志,2015,42(12):916-919.
[12]Yao CC,Tu YR,Jiang J,et al.β-elemene reverses the drug resistance of lung cancer A549/DDP cells via the mitochondrial apoptosis pathway[J].Oncol Rep,2014,31(5):2131-2138.