-
摘要:
目的 探讨乳腺癌细胞CYP1B1表达与紫杉醇耐药产生的相关性。 方法 以乳腺癌细胞系MCF-7为研究对象, 经TCDD诱导MCF-7表达CYP1B1, 观察CYP1B1表达后对紫杉醇药物的细胞毒效应的影响。将紫杉醇联合CYP1B1的拮抗剂ANF使用, 观察ANF是否可逆转CYP1B1对紫杉醇的耐药作用。采用RT-PCR测定诱导后CYP1B1 mRNA水平, 采用细胞免疫化学染色和蛋白印迹技术测定诱导后CYP1B1蛋白水平, MTS比色法测定紫杉醇对细胞增殖的抑制强度。 结果 TCDD可诱导MCF-7细胞表达CYP1B1, 其mRNA和蛋白表达水平与TCDD浓度具有明显的剂量依赖性(P < 0.05);而单独紫杉醇不能诱导MCF-7细胞表达CYP1B1。当MCF-7细胞经TCDD诱导表达CYP1B1后, 紫杉醇浓度在0.01~0.1μg/mL时, 紫杉醇对MCF-7细胞的抑制率与对照组相比明显下降(P <0.05)。使用CYP1B1的特异性拮抗剂ANF后, MCF-7细胞对紫杉醇的敏感性升高, 细胞抑制率与对照组比较无显著性差异(P>0.05)。 结论 CYP1B1表达后可导致细胞产生对紫杉醇耐药。 -
关键词:
- 人乳腺癌细胞MCF-7 /
- 2, 3, 7, 8-四氯二苯并二恶英 /
- 细胞色素P4501B1 /
- α-奈黄酮 /
- 紫杉醇耐药
Abstract:Objective To study the relevance between CYP1B1 expression and paclitaxel resistance in human breast cancer cell line. Methods Using human breast cancer cell line MCF-7 as model to observe the effect of CYP1B1 expression which induced by 2,3, 7,8-Tetrachlorodibenzo-p-dioxin (TCDD) on paclitaxel resistance. To observe if pacilitaxel resistance of MCF-7 is specifically induced by CYP1B1 expression. We treated MCF-7 with CYP1B1 specific inhibitorct-naphthoflavone combined with paclitaxel. RT-PCR was used to detect the CYP 1B 1 mRNA expression; CYP 1B 1 protein expression was detected by cell immunochemical staining and Western blotting. The cytotoxicity ofpaclitaxel was assayed by MTS method. Results TCDD can induce human breast cancer cell line MCF-7 stably express CYP1B1, CYP1B1 mRNA and protein expression have dose dependency with the concentration of TCDD(P < 0.05);Paclitaxel can not induce MCF-7 cell line express CYP 1B 1.Treated MCF-7 cell line with paclitaxel at different concentration (0.01-0.1 μg/ml), the inhibitory rate of paclitaxel on CYP1B1 expression group was reduced significantly(P < 0.05), compared with control group. This paclitaxel resistance can be reversed by 10 μmol/L ANF. Conclusion In human breast cell line MCF-7, CYP 1B 1 expression which was induced by TCDD can increase tumor cells survival and lead to paclitaxel resistance. -
图 1 不同浓度TCDD及紫杉醇诱导MCF-7后CYP1B1 mRNA的表达
Figure 1. TCDD induced expression of CYP1B1 mRNA in MCF-7
图 2 CYP1B1蛋白在MCF-7细胞中的表达
A：细胞免疫组织化学结果：TCDD诱导后，CYP1B1在细胞内的分布；B：MCF-7细胞20X光镜；C：TCDD（1.0，2.0，8.0 nmol/L）诱导下，MCF-7细胞中CYP1B1的表达（+）阳性对照（基因重组的人CYP1B1蛋白）；（-）阴性对照（无TCDD刺激的MCF-7细胞）
Figure 2. TCDD induced expression of CYP1B1 protein in MCF-7
-
[1] Van Schaik RH. Cancer treatment and pharmacogenetics of cytochrome P450 enzymes[J]. Invest New Drugs, 2005, 23(6): 513-522. doi: 10.1007/s10637-005-4019-1 [2] Rodriguez-Antona C, Ingelman-Sundberg M. CytochromeP450 pharmacogenetics and cancer[J]. Oncogene, 2006, 25(11): 1679-1691. doi: 10.1038/sj.onc.1209377 [3] Fujita K. Cytochrome P450 and anticancer drugs[J]. Curr Drug Metab, 2006, 7(1): 23-37. doi: 10.2174/138920006774832587 [4] Taylor RT, Wang F, Hsu EL, et al. Roles of Coactivator Proteins in Dioxin Induction of CYP1B1 and CYP1B1 in Human Breast Cancer Cells[J]. Toxicol Science, 2009, 107(1): 1-8. doi: 10.1093/toxsci/kfn217 [5] Cavalieri E, Rogan E. Catechol quinones of estrogens in the initiation of breast, prostate, and other human cancers: keynote lecture[J]. Ann N Y Acad Sci, 2006, 1089: 286-301. doi: 10.1196/annals.1386.042 [6] 陈悦, 邓觐云, 李张云等. 乳腺癌GYP1B1基因与ER、PR的表达关系及临床意义[J]. 江西医药, 2009, 44(12): 1176-1178. doi: 10.3969/j.issn.1006-2238.2009.12.008 [7] Hanna IH, Dawling S, Roodi R, et al. Cytochrome P450 1B1(CYP1B1) pharmacogenetic: associ-ation of polymorphisms with functional differences in estrogen hydroxylation activity[J]. Cancer Res, 2000, 60(13): 3440-3444. [8] 朱壮彦, 糜若然. 卵巢癌细胞系A2780紫杉醇体外化疗后CYP1B1基因表达差异研究[J]. 中国实用妇科与产科杂志, 2008, 24(2): 143-145. doi: 10.3969/j.issn.1005-2216.2008.02.021 [9] Sánchez-Moreno P, Boulaiz H, Ortega-Vinuesa JL, et al. Novel drug delivery system based on docetaxel-loaded nanocapsules as a therapeutic strategy against breast cancer cells[J]. Int J Sci, 2012, 13(4): 4906-4919. [10] Mathews MS, Vo V, Shih EC, et al. Photochemical internalization-mediated delivery of chemotherapeutic agents in human breast tumor cell lines[J]. J Environ Pathol Toxicol Oncol, 2012, 31(1): 49-59. doi: 10.1615/JEnvironPatholToxicolOncol.v31.i1.60 [11] Yang C, Cui ZC, Liu YJ, et al. Biological characteristics of CD55(hig); side population in breast cancer cell line MCF-7[J]. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi, 2012, 28(5): 492-496. [12] 刘移民, Yan Yan, Beverly D. Lyn-Cook. 多环芳烃受体基因与细胞色素P4501A1/1B1基因表达的调控[J]. 中国职业医学, 2003, 30(3): 10-13. doi: 10.3969/j.issn.1000-6486.2003.03.005 [13] Tokizane T, Shiina H, Igawa M, et al. Cytochrome P450 1B1 Is Overexpressed and Regulated by Hypomethylation in Prostate Cancer[J]. Clin Cancer Res, 2005, 11(16): 5793-5801. doi: 10.1158/1078-0432.CCR-04-2545 [14] Rochat B, Morsman JM, Murray GI, et al. Human CYP1B1 and anticancer agent metabolism: mechanism for tumor-specific drug inactivatin[J]? Pharmacol Exp Ther, 2001, 296(2): 537-541. [15] McFadyen M, Mcleod H, Melvin WT, et al. Cytochrome P450 CYP1B1 protein expression: a novel mechanism of anticancer drug resistance[J]. Biochem Pharmacol, 2001, 62(2): 207-212. doi: 10.1016/S0006-2952(01)00643-8 -
下载:
点击查看大图
图(3)
计量
- 文章访问数: 22
- HTML全文浏览量: 1
- PDF下载量: 1
- 被引次数: 0
