Breast cancer resistance protein （Bcrp） and the testis--an unexpected turn of events

Breast cancer resistance protein （Bcrp） is an ATP-dependent efflux drug transporter. It has a diverse spectrum of hydrophilic and hydrophobic substrates ranging from anticancer, antiviral and antihypertensive drugs, to organic anions, antibiotics, phytoestrogens （e.g., genistein, daidzein, coumestrol）, xenoestrogens and steroids （e.g., dehydroepiandrosterone sulfate）. Bcrp is an integral membrane protein in cancer and normal cells within multiple organs （e.g., brain, placenta, intestine and testis） that maintains cellular homeostasis by extruding drugs and harmful substances from the inside of cells. In the brain, Bcrp is a major component of the blood- brain barrier located on endothelial cells near tight junctions （TJs）. However, Bcrp is absent at the Sertoli cell blood-testis barrier （BTB）; instead, it is localized almost exclusively to the endothelial TJ in microvessels in the interstitium and the peritubular myoid cells in the tunica propria. Recent studies have shown that Bcrp is also expressed stage specifically and spatiotemporally by Sertoli and germ cells in the seminiferous epithelium of rat testes, limited only to a testis-specific cell adhesion ultrastructure known as the apical ectoplasmic specialisation （ES） in stage VI-early VIII tubules. These findings suggest that Bcrp is equipped by late spermatids and Sertoli cells to protect late-stage spermatids completing spermiogenesis. Furthermore, Bcrp was found to be associated with F （filamentous）-actin and several actin regulatory proteins at the apical ES and might be involved in the organisation of actin filaments at the apical ES in stage VII-VIII tubules. These findings will be carefully evaluated in this brief review.

High expression of autophagy-related gene EIF4EBP1 could promote tamoxifen resistance and predict poor prognosis in breast cancer

BACKGROUND Breast cancer(BC) remains a public health problem. Tamoxifen(TAM) resistance has caused great difficulties for treatment of BC patients. Eukaryotic translation initiation factor 4E binding protein 1(EIF4EBP1) plays critical roles in the tumorigenesis and progression of BC. However, the expression and mechanism of EIF4EBP1 in determining the efficacy of TAM therapy in BC patients are still unclear.AIM To investigate the expression and functions of EIF4EBP1 in determining the efficacy of TAM therapy in BC patients.METHODS High-throughput sequencing data of breast tumors were downloaded from the Gene Expression Omnibus database. Differential gene expression analysis identified EIF4EBP1 to be significantly upregulated in cancer tissues. Its prognostic value was analyzed. The biological function and related pathways of EIF4EBP1 was analyzed. Subsequently, the expression of EIF4EBP1 was determined by real-time reverse transcription polymerase chain reaction and western blotting. Cell Counting Kit-8 assays, colony formation assay and wound healing assay were used to understand the phenotypes of function of EIF4EBP1.RESULTS EIF4EBP1 was upregulated in the TAM-resistant cells, and EIF4EBP1 was related to the prognosis of BC patients. Gene Set Enrichment Analysis showed that EIF4EBP1 might be involved in Hedgehog signaling pathways. Decreasing the expression of EIF4EBP1 could reverse TAM resistance, whereas overexpression of EIF4EBP1 promoted TAM resistance.CONCLUSION This study indicated that EIF4EBP1 was overexpressed in the BC and TAM-resistant cell line, which increased cell proliferation, invasion, migration and TAM resistance in BC cells.

Breast Cancer Resistance Protein Expression and 5-Fluorouracil Resistance

Objective To filtrate breast cancer resistance protein （BCRP）-mediated resistant agents and to investigate clinical relationship between BCRP expression and drug resistance. Methods MTT assay was performed to filtrate BCRP-mediated resistant agents with BCRP expression cell model and to detect chemosensitivity of breast cancer tissue specimens to these agents. A high performance liquid chromatography （HPLC） assay was established, and was used to measure the relative dose of intracellular retention resistant agents. RT-PCR and immunohistochemistry （IHC） were employed to investigate the BCRP expression in breast cancer tissue specimens. Results MTT assay showed that the expression of BCRP increased with the increasing resistance of 5-fluorouracil （5-Fu） （P〈0.05, n=3） in the cell model, while HPLC assay indicated that the intracellular retention dose of 5-Fu was significantly correlated with the expression of BCRP （t=-0.897, P〈0.05, n=3）. A total of 140 breast cancer tissue specimens were collected. BCRP-positive expression was detected in forty-seven specimens by both RT-PCR and IHC. As shown by MTT assay subsequently, the resistance index （RI） of 47 BCRP-positive breast cancer tissue specimens to 5-Fu was 7-12 times as high as that of adjacent normal tissue samples. BCRP expression was related to 5-Fu resistance （R2=0.8124, P〈0.01）. Conclusion Resistance to 5-Fu can be mediated by BCRR Clinical chemotherapy for breast cancer patients can be optimized based on BCRP-positive expression.

Effect of Ursolic Acid on Breast Cancer Resistance Protein-mediated Transport of Rosuvastatin In Vivo and Vitro

Objective To evaluate whether ursolic acid can inhibit breast cancer resistance protein(BCRP)-mediated transport of rosuvastatin in vivo and in vitro. Methods Firstly, we explored the pharmacokinetics of 5-fluorouracil(5-FU, a substrate of BCRP) in rats in the presence or absence of ursolic acid. Secondly, we studied the pharmacokinetics of rosuvastatin in rats in the presence or absence of ursolic acid or Ko143(inhibitor of BCRP). Finially, the concentration-dependent transport of rosuvastatin and the inhibitory effects of ursolic acid and Ko143 were examined in Madin-Darby Canine Kidney(MDCK) Ⅱ-BCRP421CC(wild type) cells and MDCKⅡ-BCRP421AA(mutant type) cells. Results As a result, significant changes in pharmacokinetics parameters of 5-FU were observed in rats following pretreatment with ursolic acid. Both ursolic acid and Ko143 could significantly affect the pharmacokinetics of rosuvastatin. The rosuvastatin transport in the BCRP overexpressing system was increased in a concentration-dependent manner. However, there was no statistical difference in BCRP-mediated transport of rosuvastatin betweent the wild type cells and mutant cells. The same as Ko143, ursolic acid inhibited BCRP-mediated transport of rosuvastatin in vitro. Conclusion Ursolic acid appears to be a potent modulator of BCRP that affects the pharmacokinetic of rosuvastatin in vivo and inhibits the transport of rosuvastatin in vitro.

Differential expression of breast cancer-resistance protein,lung resistance protein,and multidrug resistance protein 1 in retinas of streptozotocin-induced diabetic mice

AIM：To investigate the altering expression profiles of efflux transporters such as breast cancer-resistance protein（BCRP）,lung resistance protein（LRP）,and multidrug resistance protein 1（MDR1） at the inner blood-retinal barrier（BRB） during the development of early diabetic retinopathy（DR） and/or aging in mice.METHODS：Relative m RNA and protein expression profiles of these three efflux transporters in the retina during the development of early DR and/or aging in mice were examined.The differing expression profiles of Zonula occludens 1（ ZO-1） and vascular endothelial growth factor-A（ VEGFA） in the retina as well as the perfusion characterization of fluorescein isothiocyanate（FITC）-dextran and Evans blue were examined to evaluate the integrity of the inner BRB.RESULTS：There were significant alterations in these three efflux transporters＇ expression profiles in the m RNA and protein levels of the retina during the development of diabetes mellitus and/or aging.The development of early DR was confirmed by the expression profiles of ZO-1 and VEGFA in the retina as well as the compromised integrity of the inner BRB.CONCLUSION：The expression profiles of some efflux transporters such as BCRP,LRP,and MDR1 in mice retina during diabetic and/or aging conditions are tested,and the attenuated expression of BCRP,LRP,and MDR1 along with the breakdown of the inner BRB is found,which may be linked to the pathogenesis of early DR.

Expression of MxA Protein in Triple Negative Breast Cancer and its Relationship with Prognosis

Objective:To explore the expression of human myxovirus resistance protein A(MxA)in triple negative breast cancer(TNBC)and its relationship with prognosis.Methods:144 cases of TNBC confirmed by pathology before or after surgery from January 2014 to January 2017 in the First Central Hospital of Baoding City were retrospectively collected.Western blotting was used to detect the expression of MxA protein in TNBC and adjacent breast tissues.According to the expression of MxA protein,144 TNBC patients were divided into low MxA protein expression group(n=91)and MxA protein high expression group(n=53)for subsequent comparison of prognosis of patients in between these two groups.Results:The expression of MxA protein in TNBC tissue was lower than that of adjacent breast tissue,and the difference was statistically significant(P<0.05).The patients in high MxA expression group had higher loco-regional recurrence-free rate,disease-free survival(DFS)rate,and overall survival(OS)rate than those in low MxA expression group for 3 years.On the other hand,the distant metastasis rate was lower in the high expression group compared to that in the low MxA expression group,and the difference was statistically significant(P<0.05).Conclusion:In triple-negative breast cancer,high MxA expression is a potential predictor of TNBC prognosis.

EFFECTS OF NEOADJUVANT CHEMOTHERAPY ON MDR1 AND MRP GENE EXPRESSION IN PRIMARY BREAST CANCER

Objective: To investigate the effects of neoadjuvant chemotherapy on the expression of drug resistance genes, multidrug resistance-1 (MDR1) and multidrug resistance-associated protein (MRP), in patients with primary breast cancer. Methods: MDR1 and MRP expression were detected by semi-quantitative RT-PCR in 20 patients with primary breast cancer, before and after chemotherapy. Results: Before chemotherapy, MDR1 and MRP expression can be detected in 15 cases (75%) and 18 cases (90%) respectively. After chemotherapy, expression of MDR1 is not significantly different from that before chemotherapy, but expression of MRP is significantly different from that before chemotherapy. Conclusion: Expression of drug resistance gene MRP, but not MDR1, is enhanced in patients with primary breast cancer submitted to neoadjuvant chemotherapy.

miR-137通过调控血红素铁输出蛋白FLVCR、BCRP对大鼠I/R损伤的保护作用

目的探讨大鼠脑缺血/再灌注(I/R)后血红素铁输出蛋白猫白血病病毒C亚组受体(FLVCR)、乳腺癌抗性蛋白(BCRP)的表达变化及miR-137对其表达的调控作用。方法选取SD大鼠60只,随机分为假手术(SO)组、模型(MCAO/R)组、miR-137模拟物(Mimic)组、miR-137模拟对照(Mimic对照)组、miR-137抑制物(Inhibitor)组、miR-137抑制对照(Inhibitor对照)组,每组10只。采用线栓法制作大鼠脑缺血再灌注(MCAO/R)模型,分别于再灌注后12、24 h取材。通过qRT-PCR测定大鼠脑组织FLVCR、BCRP及miR-137的mRNA表达,免疫组化检测FLVCR、BCRP蛋白的表达水平。结果MCAO/R组大鼠神经功能缺损评分显著升高(P<0.01),Mimic组评分显著下降(P<0.01),Inhibitor组评分显著升高(P<0.05)。MCAO/R组miR-137 mRNA表达水平显著降低(P<0.05),与Mimic对照组同时间比较,Mimic组miR-137 mRNA表达水平显著升高(P<0.01),且24 h升高更显著(P<0.05);与Inhibitor对照组同时间比较,Inhibitor组miR-137 mRNA表达水平显著降低(P<0.05),且24 h降低更显著(P<0.05)。与SO组比较,MCAO/R组FLVCR mRNA及蛋白表达水平均显著降低(P<0.05);与Mimic对照组比较,Mimic组FLVCR mRNA及蛋白表达水平在12 h显著升高(P<0.05),24 h最高(P<0.05);与Inhibitor对照组比较,Inhibitor组FLVCR mRNA及蛋白表达水平降低(P<0.01),在24 h降低更明显(P<0.05)。与SO组比较,MCAO/R组BCRP mRNA及蛋白表达水平显著降低(P<0.05);与Mimic对照组比较,Mimic组BCRP mRNA及蛋白表达水平在12 h显著升高(P<0.01),24 h最高(P<0.05);与Inhibitor对照组比较,Inhibitor组BCRP mRNA及蛋白表达水平显著降低(P<0.01)。结论miR-137过表达可能促进FLVCR和BCRP的表达以保护大鼠脑缺血再灌注损伤。

BCRP基因在乳腺癌组织中的表达及意义

目的 研究乳腺癌耐药蛋白 (BCRP)基因在乳腺癌组织中的表达水平 ,探讨其与肿瘤病理分期、淋巴结转移的关系。方法 采用实时荧光定量 RT- PCR检测 5 1例乳腺癌患者癌组织和癌旁组织中 BCRP基因表达。结果 BCRP基因在乳腺癌组织中的阳性表达率、表达水平均显著高于癌旁组织 (P<0 .0 1、<0 .0 5 ) ;BCRP基因在 、 、 期乳腺癌组织中的表达水平无显著性差异 (P>0 .0 5 ) ;有淋巴结转移者的 BCRP基因表达水平显著高于无淋巴结转移者 (P<0 .0 5 )。结论 BCRP基因表达水平与肿瘤病理分期无关 ,与淋巴结转移有关 ,BCRP基因过表达可能在乳腺癌的多药耐药中起较重要的作用。

CK5、CK8和BCRP在人乳腺病变组织中的表达及其意义

目的:探讨细胞角蛋白5(cytokeratin 5,CK5)、CK8和乳腺癌耐药蛋白(breast cancer resistance protein,BCRP)在乳腺上皮良、恶性增生病变中的表达及意义。方法:应用免疫组织化学法分别检测89例正常乳腺、乳腺增生、导管上皮非典型增生、导管原位癌和浸润性导管癌组织中CK5、CK8及BCRP的表达情况,分析CK5、CK8及BCRP的表达与乳腺病变组织分化程度之间的关系。结果:在正常乳腺、乳腺增生、导管上皮非典型增生、导管原位癌和浸润性导管癌组织中CK5和CK8的阳性表达率逐渐下降,BCRP的阳性表达率则逐渐上升。CK5和CK8在乳腺良性增生组织(乳腺增生和导管上皮非典型增生)中的阳性表达率明显高于乳腺癌(导管原位癌和浸润性导管癌)组织(P<0.01),BCRP在乳腺癌组织中的表达明显高于乳腺良性增生组织(P<0.01)。在乳腺浸润性导管癌中,CK5阳性表达主要集中在外层尚未受侵袭的基层细胞中,肿瘤细胞中没有表达或很少表达;CK8则主要在肿瘤细胞中表达,基层细胞中完全不表达;66.7%的浸润性导管癌患者呈CK5-/CK8+表型。结论:CK5、CK8和BCRP在不同乳腺病变组织中的表达不同,3者联合检测可能有助于乳腺病变的病理学诊断。

MDR1、BCRP和LRP基因在乳腺癌组织中的表达及其意义

目的探讨多药耐药基因(MDR1)、乳腺癌耐药蛋白(BCRP)及肺耐药蛋白(LRP)mRNA在乳腺癌组织中的表达及其相互关系,分析它们与乳腺癌临床病理特征的关系。方法采用RT-PCR方法检测2007年1月至2007年12月在宁夏医科大学附属医院手术切除的42例乳腺癌组织、42例癌旁组织及50例乳腺良性病变中MDR1、BCRP和LRP mRNA的表达。结果乳腺癌组织中MDR1、BCRP、LRP mRNA的阳性表达率分别为40.47%、38.09%及61.90%,与癌旁组织及乳腺良性病变组织相比阳性表达率均有统计学差异(P<0.05)。MDR1mRNA表达与绝经状况呈完全正相关(r=0.398,P<0.01),与腋窝淋巴结状况呈完全正相关(r=0.398,P<0.01);BCRP mRNA表达与腋窝淋巴结状况呈正相关(r=0.355,P<0.05);LRP mRNA表达与绝经状况、年龄、腋窝淋巴结状况及肿瘤大小之间均无相关性(P>0.05);MDR1mRNA和BCRP mRNA表达呈正相关(r=0.652,P<0.01),LRP mRNA与MDR1mRNA表达无相关性(r=0.147,P>0.05);LRP mRNA和BCRP mRNA表达无相关性(r=0.111,P>0.05)。2个耐药基因共表达率为47.61%,2种或3种耐药基因共表达率为61.89%。结论乳腺癌组织中存在耐药基因MDR1、BCRP和LRP的表达,单基因和多基因协同作用,以多基因共表达为主;检测MDR1和BCRP基因表达水平可辅助临床判断乳腺癌患者的预后。

耐药蛋白P-gp、BCRP及LRP在乳腺癌原发灶和腋淋巴结转移灶中表达的比较

目的研究P-糖蛋白(P-glycoprotein,P-gp)、乳腺癌耐药蛋白(breast cancer drug resistance protein,BCRP)和肺耐药蛋白(pulmonary resistance protein,LRP)在乳腺癌原发灶和腋淋巴结转移灶中表达的差异。方法采用免疫组织化学染色法检测126例乳腺癌患者的原发灶和66例腋淋巴结转移灶中P-gp、BCRP、LRP的表达。结果 (1)P-gp、BCRP及LRP在乳腺癌原发灶中的阳性表达率分别为41.27%(52/126)、38.89%(49/126)、65.87%(83/126),在腋淋巴结转移灶中阳性表达率分别为59.09%(39/66)、63.64%(42/66)、60.61%(40/66)。P-gp、BCRP在乳腺癌淋巴结转移灶中的表达高于原发灶(P<0.05),LRP在原发灶和淋巴结转移灶之间的表达差异无统计学意义。(2)P-gp、BCRP在乳腺癌原发灶和腋淋巴结转移灶中表达差异没有统计学意义(P<0.01),Kappa值分别为0.276、0.356;LRP在原发灶和淋巴结转移灶中的表达差异没有统计学意义(P>0.05)。(3)乳腺癌原发灶中2个耐药蛋白共表达率为35.71%(45/126),3个耐药蛋白同时表达的阳性率为15.08%(19/126),有2个或3个耐药蛋白共表达率为50.79%(64/126),高于单独阳性表达率33.33%(42/126,P<0.05)。腋淋巴结转移灶中2个耐药蛋白共表达率为53.03%(35/66),高于单独阳性表达率(27.27%[18/66],P<0.05);3个耐药蛋白同时表达的阳性率为16.67%(11/66),有2个或3个耐药蛋白共表达率为69.70%(46/66),高于单独阳性表达率(P<0.01)。腋淋巴结转移灶中2个耐药蛋白共表达率及2个或3个耐药蛋白共表达率均高于乳腺癌原发灶中的表达(P<0.05)。(4)Kaplan-Meier生存分析结果表明,乳腺癌腋淋巴结转移灶中P-gp、BCRP及LRP阳性表达者5年总生存期较原发灶耐药蛋白阳性者低(P<0.05)。结论在乳腺癌原发灶和腋淋巴结转移灶中的P-gp、BCRP表达差异有统计学意义,LRP表达则没有明显差异;多个耐药蛋白共表达协同作用为耐药的主要特征,腋淋巴结转移灶中可能具有更强的耐药性。

BCRP过表达对乳腺癌细胞增殖和凋亡的影响

目的探讨乳腺癌耐药蛋白(BCRP)基因过表达对人乳腺癌细胞MDA-MB-435S增殖和凋亡的影响。方法取对数生长期的MDA-MB-435S细胞,将其随机分为三组,阳性转染组将真核表达质粒pc DNA3.1/BCRP经双酶切和测序鉴定后转染MDA-MB-435S细胞,阴性对照组同步转染2μg pc DNA3.1的空质粒,空白对照组用2 m L原培养液正常培养。采用RT-PCR法和Western blotting法检测BCRP mRNA和蛋白表达。转染后48 h,每组再分设7个丝裂霉素(MMC)浓度(0.01、0.10、2.50、5.00、10.00、20.00、40.00μmol/L)组,每个浓度组3个复孔,加入相应浓度的MMC 10μL,采用CCK-8法检测细胞增殖情况;流式细胞术分析细胞凋亡情况。结果重组真核表达质粒pc DNA3.1/BCRP经双酶切和测序证实该质粒构建正确;阳性转染组转染24、48、72 h后,MDA-MB-435S细胞中BCRP mRNA和蛋白表达均明显增加(P均<0.05);与阴性对照组比较,阳性转染组转染pc DNA3.1/BCRP 48 h后,细胞凋亡率无统计学差异(P>0.05);CCK-8结果显示,化疗药物MMC作用24 h后,与阴性对照组比较,阳性转染组转染pc DNA3.1/BCRP 48h后细胞存活率显著增加(P<0.05)。结论 BCRP过表达可促进人乳腺癌细胞MDA-MB-435S增殖,但对细胞凋亡无明显影响。

MCF-7/BCRP细胞系的建立及其生物学特征

目的:建立表达乳腺癌耐药蛋白(BCRP)的耐药细胞系MCF-7/BCRP。方法:提取MCF-7细胞的总RNA,设计BCRP基因上下游引物,用RT-PCR法克隆出BCRP基因全长并连接到真核表达载体pcDNA3.1穴-雪上,转染MCF-7细胞后以G418筛选细胞。采用米托蒽醌外排实验、细胞毒性实验、细胞群体倍增时间、免疫荧光法鉴定构建的耐药细胞系。结果:MCF-7/BCRP对米托蒽醌耐药指数增加至14.72;外排米托蒽醌的作用增强;细胞群体倍增时间较亲代细胞延长;MCF-7/BCRP细胞能表达一定量的BCRP。结论:MCF-7/BCRP细胞能够表达BCRP并具有BCRP的耐药表型。

不同分子亚型乳腺癌组织中BCRP、MRP1和MDR1的表达及其临床意义

目的根据乳腺癌组织中ER、PR、Her-2和Ki-67的水平对乳腺癌进行分子分型,观察不同分子亚型中乳腺癌耐药蛋白(BCRP)、多药耐药相关蛋白1(MRP1)和多药耐药蛋白1(MDR1/P-gp)的表达,探讨这3种多药耐药蛋白在不同分子亚型乳腺癌中的表达及临床意义。方法用免疫组织化学方法检测79例乳腺癌组织中ER、PR、Her-2、Ki-67、BCRP、MRP1和MDR1分子的表达水平,分析3种多药耐药蛋白与乳腺癌分子亚型及ER、Her-2、Ki-67分子表达间的关系。结果乳腺癌组织中BCRP、MRP1和MDR1阳性表达率分别为77.2%、38.0%、7.6%。MRP1在luminal A型高表达(68.4%)(P<0.05);BCRP和MDR1在luminal A型、luminal B型、Her-2过表达型和三阴性型乳腺癌的阳性表达率比较,差异均无统计学意义(P>0.05)。BCRP和MRP1的表达呈正相关(r=0.301,P<0.05),BCRP和MDR1、MRP1和MDR1的表达无相关性。MRP1与ER的表达呈正相关(r=0.290,P<0.05),与Her-2、Ki-67的表达无相关性;BCRP和MDR1分别与ER、Her-2、Ki-67的表达均无相关性。结论BCRP、MRP1和MDR1在乳腺癌组织中具有不同程度的表达,MRP1可作为预测乳腺癌尤其是luminal A型乳腺癌内源性耐药的指标。

乳腺癌组织中BCRP的表达与新辅助化疗疗效的关系

目的探讨原发性乳腺癌组织中乳腺癌耐药蛋白(breast cancer resistance protein,BCRP)表达与新辅助化疗疗效的相关性。方法采用免疫组织化学MaxVisionTM一步法检测84例原发性乳腺癌组织中BCRP的表达,结合新辅助化疗后临床疗效及术后病理组织学Miller&Payne(MP)分级,分析BCRP的表达与新辅助化疗疗效及相关病理指标的相互关系。结果 (1)原发性乳腺癌组织中BCRP的阳性表达率为71.43%。(2)BCRP的表达水平与新辅助化疗临床疗效有关,新辅助化疗后获得cCR组患者BCRP表达水平明显低于SD+PD组和cPR组,三组间BCRP表达水平的差异有统计学意义(χ2=9.779,P=0.008)。(3)BCRP的表达水平与新辅助化疗后组织学反应有关,组织学显著反应组(病理反应4~5级)BCRP的表达水平明显低于组织学非显著反应组(病理反应1~3级),差异有统计学意义(χ2=8.649,P=0.003)。(4)BCRP的表达水平与新辅助化疗后残余阳性腋窝淋巴结的个数正相关(r=0.518,P=0.000)。结论 BCRP在原发性乳腺癌组织中有一定程度的表达,BCRP的表达水平可以预测新辅助化疗的临床疗效及组织学反应,BCRP的表达水平与新辅助化疗后残余阳性腋窝淋巴结的个数正相关,BCRP可以作为判断患者化疗效果及预后指标之一。

大鼠脑缺血后海马CA2区FLVCR和BCRP的表达及益气活血法干预

目的研究脑缺血后血红素铁转运蛋白人类猫白血病C亚类病毒受体(feline leukemia virus subgroup C receptor,FLVCR)及胸腺癌抵抗蛋白(breast cancer resistance protein,BCRP)随时间表达及益气活血法脑泰方(黄芪、川芎、地龙、僵蚕)提取物干预对FLVCR及BCRP表达的影响。方法实验分两步进行,第一部分实验:随机将SD大鼠分为2 h、6 h、12 h、24 h、72 h组,采用大脑中动脉栓塞法(middle cerebral artery occlusion,MCAO)行大鼠局灶性脑缺血模型制备,分别在术后2 h、6 h、12 h、24 h、72 h各时间点取材,通过免疫组织化学及RT-PCR检测FLVCR和BCRP及FLVCR mRNA和BCRP mRNA随时间表达的变化。第二部分实验,随机将SD大鼠分为假手术组、模型组、脑泰方低、中、高剂量组(3、9、27 g/kg)。各组大鼠预处理藻胃给药连续3天,MCAO模型制备术后连续藻胃给药3天,每日1次。术后3天取材,电镜检测海马神经元内铁样颗粒分布,免疫组织化学及RT-PCR检测FLVCR和BCRP及FLVCR mRNA和BCRP mRNA表达。结果 72 h组FLVCR表达明显降低(P<0.05),BCRP的表达2 h、6h较高,12 h、24 h、72 h表达明显降低(P<0.05);电镜下脑泰方各剂量组神经元内铁样颗粒聚集较模型组减少。脑泰方高剂量组FLVCR及FLVCR mRNA表达明显增加(P<0.05),BCRP表达无明显改变。结论大鼠脑缺血后第72 h FLVCR表达下降,益气活血中药脑泰方提取物干预可增加脑缺血后FLVCR的表达,促进血红素铁的外排,可能是治疗脑缺血损伤的新机制。

乳腺癌组织中ALDH1及BCRP的表达状况与乳腺癌病理特征的相关性分析

目的探讨乳腺癌患者癌组织及腋窝淋巴结中乙醛脱氢酶1(ALDH1)以及乳腺癌耐药蛋白(BCRP)的表达状况与乳腺癌病理特征的关系。方法选取2013年1月~2016年3月在四川大学华西医院治疗的乳腺癌患者共84例,同时选取距肿瘤边缘大于1 cm正常乳腺组织(癌旁正常组织)作为对照,采用免疫组化法检测ALDH1和BCRP蛋白的表达。结果乳腺癌组织中ALDH1和BCRP蛋白表达率明显高于癌旁正常组织(P<0.01);腋窝淋巴结有转移患者癌组织中ALDH1和BCRP蛋白表达阳性率明显高于无腋窝淋巴结转移患者(P<0.05);雌激素受体(ER)阳性患者乳腺癌组织中BCRP蛋白表达阳性率明显高于ER阴性患者(P<0.05);而乳腺癌组织中ALDH1和BCRP蛋白表达与患者年龄、绝经情况、肿瘤大小、组织分级、TNM分期和孕激素受体(PR)均无相关性(P>0.05);乳腺癌患者ALDH1和BCRP蛋白的表达也无相关性(rs=-0.092,P>0.05)。结论 ALDH1和BCRP蛋白在乳腺癌发生发展中有一定作用,可能参与了肿瘤的侵袭、转移等过程。

乳腺癌中医辨证分型与BCRP表达的相关性分析

目的探讨乳腺癌患者的中医辨证分型与乳腺癌耐药蛋白(BCRP)表达的相关性。方法对80例乳腺癌患者进行术前中医辨证分型,其中血瘀证23例、肝郁证30例、脾虚证17例、肾阴虚证10例,对术后标本进行BCRP检测,并进行分析。结果乳腺癌患者术前辨证为血瘀证者BCRP表达的阳性率高于其他证型,差异有统计学意义(P<0.05)。结论乳腺癌术前中医辨证与肿瘤BCRP表达有相关性,血瘀证者BCRP表达的阳性率最高,预后差。

绵萆薢水提物及其主要成分薯蓣皂苷对肾小管上皮细胞膜乳腺癌耐药蛋白Bcrp的调控

目的探讨绵萆薢水提物及其主要成分薯蓣皂苷对分布于大鼠肾小管上皮细胞膜的乳腺癌耐药蛋白(Bcrp/Abcg2)的调控。方法雄性Wistar大鼠随机分为正常组、模型对照组、模型组、别嘌醇组(2.5 mg/kg)、绵萆薢水提物和薯蓣皂苷高、中、低剂量(638.43、319.22、159.61 mg/(kg·d))组(n=8)。用氧嗪酸钾联合乙胺丁醇制备高尿酸血症大鼠模型,从造模第1天起连续灌胃给予生理盐水和不同剂量的绵萆薢水提物或薯蓣皂苷,1次/天,至给药第10天,采用HPLC测定血尿酸及尿酸排泄量,并通过Western Blot检测Bcrp蛋白表达水平,采用离体肾灌流实验,考察Bcrp抑制剂新生霉素对大鼠累积尿酸排泄量的影响。结果模型组大鼠血尿酸水平显著升高,尿尿酸排泄减少,其肾脏组织中Bcrp的蛋白表达水平均显著降低,高剂量绵萆薢水提物及薯蓣皂苷可显著降低高尿酸血症大鼠的血清尿酸水平,增加尿尿酸排泄量,并增加肾脏组织中Bcrp蛋白表达水平。此外,抑制Bcrp可显著降低尿尿酸排泄量。结论绵萆薢水提物及高剂量薯蓣皂苷可增加Bcrp的表达,这可能是其抗高尿酸血症的一个重要机制。