Expression and Prognostic Significance of Multidrug Resistance Associated Protein (MRP) Gene in Non-small Cell Lung Cancer by in Site Hybridization

Objective: To study on the effect of MRP gene overexpression on prognosis of patients with non-small lung cancer (NSCLC). Methods: Paraffin-embedded tissues from 47 cases of NSCLC who had undergone radical tumor resection were examined for expression of MRP gene mRNA by in situ hybridization using labelled digoxigenin probes combined with immunohistochemistry. All the patients were retrospectively followed-up. Results: All of the 47 lung cancer specimens were found to have overexpression of MRP gene mRNA. It was significantly correlated with patients' survival time, response to chemotherapy, recurrence or metastases after surgery, but was not correlated with histology, tumor size, node status, TNM stage, degree of differentiation, age and sex. Conclusion: Overexpression of MRP gene is a marker of prognostic significance in patients with NSCLC.

EXPRESSION AND REVERSION OF DRUG RESISTANCE-AND APOPTOSIS-RELATED GENES OF A DDP-RESISTANT LUNG ADENOCARCINOMA CELL LINE

Objective: To investigate the co-expression of drug resistance- and apoptosis-related genes of cisplatin (CDDP)-selected lung adenocarcinoma cell line A 549 DDP for compared to the parental cell line A549, and reverse of drug resistance by antisense s-oligodeoxynucleotides (S-ODNs) of differentially expressed genes. Methods: Sense and antisense S-ODN were transferred into A 549 DDP cells by lipofectin. The expression of drug resistance and apoptosis related genes was examined by RT-PCR, immunocytochemistry and flow cytometry, respectively. Apoptostic cells were identified by DNA electrophoresis and terminal deoxynucleotidyl transferase (TdT)-mediated biotin dUTP nick end-labeling(TUNEL). Drug resistance of tumor cells was detected by a cell viability (MTT) assay. Results: The expression of bcl-2 was positive and that of multidrug resistance-associated protein (MRP) at mRNA and protein level was increased in A 549 DDP compared to A549 cells. MDR1, c-myc and topoisomeras II (TOPO II) were similarly co-expressed in two cell lines. Both cell lines were negative for c-erbB-2 expression. In A 549 DDP cells, the expression of bcl-2 and MRP was significantly inhibited by their respective antisense S-ODNs. Antisense S-ODNs could also decrease significantly drug resistance of A 549 DDP cells to CDDP by promoting cell apoptosis. Conclusion: Both intrinsic and acquired drug resistance were involved in co-expression of multiple MDR-related genes in lung adenocarcinoma. Cooperation of bcl-2 and MRP genes appeared to play an important action to confer the resistance of A 549 DDP cells to CDDP. Their antisense S-ODNs are responsible for the decrease of drug resistance of this cell line by promoting apoptosis.

Reversal effect of recombinant human Endostatin on cisplatin resistance in A549/DDP human lung adenocarcinoma cells in vitro

Objective： Recombinant human Endostatin （rh-Endostatin, YH-16） can reverse cisplatin resistance in A549/DDP cells. However, the possible effect of rh-Endostatin in reversing DDP-resistance in A549/DDP cells and the mechanism are needed to be investigated. Methods： Lung adenocarcinoma cell line A549 and its DDP-resistant cell line A549/DDP were treated with DDP and/or recombinant human Endostatin. Difference in drug resistance was analyzed between different regi- mens and between different cell lines after a 72 h-treatment in vitro. And below the non-cytotoxic concentration of rh-End- ostatin, the possibility of rh-Endostatin in reversing DDP-resistance in A549/DDP was evaluated. The resistance protein which was detected in the study included P glycoprotein （P-gp） and topoisomerase II （Topo-II）. Results： Rh-Endostatin below 400 IJg/mL showed no cytotoxicity in either A549 or A549/DDP after 72 h-treatment with it. The inhibited concentration of 50% （IC50） observed for DDP was （0.79 _＋ 0.05） IJg/mL in A549 and （13.2 ＋ 1.1） in A549/DDP respectively. IC50 was reduced to 2.57 ＋ 0.05 #g/mL in A549/DDP treated by rh-Endostatin below the non-cytotoxic concentrations in combination with DDP, with a reversal fold （RF） of 5.14 and a relative reversal rate of 85.6%. Apoptotic rates were 2.01%, 13.47% and 29.26% re- spectively for cells treated with rh-Endostain, DDP, and the combination. The rate of the A549/DDP control group was 0.99%. The expression level of P-gp or Topo-II was higher in A549/DDP cells than in A549 cells. Rh-Endostatin may partially reverse DDP-resistance in A549/DDP cells in vitro, with a probable mechanism related to lowering expression of P-gp and Topo-II. Conclusien： Rh-Endostatin of non-cytotoxic dose partially reversed cisptatin resistance in cisplatin-resistant human lung adenocarcinoma cell line A549/DDP. Rh-Endostatin reversed the resistance of A549/DDP cells to DDP, which may be related to decreased protein expression of P-gp and Topo-II in A549/DDP cells.

Bioinformatics Identification of ZNFs/LINC00520/miR-181d/BCL2 Axis as a Novel Network in Cisplatin-Resistant Lung Adenocarcinoma Cells

Background: Resistance to cisplatin (DDP) leads to poor prognosis in patients with Lung Adenocarcinoma (LUAD) and limits its clinical application. It has been confirmed that autophagy promotes chemoresistance and, therefore, novel strategies to reverse chemoresistance by regulating autophagy are desperately needed. Methods: The differentially expressed lncRNAs (DElncRNAs), miRNAs (DEmiRNAs), and mRNAs (DEmRNAs) between A549 and A549/DDP cell lines were identified using the limma package in R, after gene expression profiles were obtained from Gene Expression Omnibus (GEO) database. By combining Autophagy-Related Genes (ARGs) from Human Autophagy Database (HADb), the interactions lncRNA-miRNAs and the interactions miRNAs-mRNAs respectively predicted by miRcode and miRDB/Targetscan database, the autophagy-related ceRNA network was constructed. Then, extraction of ceRNA subnetwork and Cox regression analyses were performed. A prognosis-related ceRNA subnetwork was constructed, and the upstream Transcription Factors (TFs) regulating lncRNAs were predicted by the JASPAR database. Finally, the expression patterns of candidate genes were further verified by quantitative real-time polymerase chain reaction (qRT-PCR) experiments. Results: A total of 3179 DEmRNAs, 180 DEmiRNAs, and 160 DElncRNAs were identified, and 35 DEmRNAs were contained in the HADb. Based on the ceRNA hypothesis, we established a ceRNA network, including 10 autophagy-related DEmRNAs, 9 DEmiRNAs, and 14 DElncRNAs. Then, LINC00520, miR-181d, and BCL2 were identified to construct a risk score model, which was confirmed to be a well-predicting prognostic factor. Furthermore, 5 TF ZNF family members were predicted to regulate LINC00520, whereas the RT-PCR results showed that the 5 ZNFs were consistent with the bioinformatics analysis. Finally, a ZNF regulatory LINC00520/miR-181d/BCL2 ceRNA subnetwork was constructed. Conclusions: An ZNFs/LINC00520/miR-181d/BCL2 axis as a novel network in DDP-resistant LUAD has been constructed successfully, which may provide potential therapeutic targets for LUAD.

JNJ-26481585对人肺腺癌培美曲塞耐药裸鼠移植瘤的抑制作用及其机制探讨

目的:探讨JNJ-26481585对人肺腺癌培美曲塞耐药裸鼠移植瘤的抑制作用及其抗癌机制。方法:构建人肺腺癌A549/培美曲塞耐药细胞(A549/PEM)的移植瘤裸鼠模型,随机分为模型组、培美曲塞组、JNJ组、联合组(培美曲塞+JNJ),第28天治疗观察结束时处死裸鼠,剥离肿瘤组织标本待测。测定4组裸鼠肿瘤体积与体质量,计算抑瘤率,观察各治疗组药物对荷瘤裸鼠的毒副反应,TUNEL方法检测4组移植瘤细胞凋亡情况的差异,qRT-PCR检测4组移植瘤细胞FAM96A、Bax、Bcl-2基因mRNA表达水平差异,Western blot检测4组移植瘤细胞FAM96A、Bax、Bcl-2蛋白表达的差异。结果:联合组的肿瘤体积、体质量低于JNJ组,JNJ组的肿瘤体积、体质量低于培美曲塞组,联合组的抑瘤率高于JNJ组,JNJ组抑瘤率高于培美曲塞组(均P<0.05)。联合组的细胞凋亡率高于JNJ组,JNJ组细胞凋亡率高于培美曲塞组(均P<0.05)。联合组肿瘤细胞的FAM96A、Bax表达水平高于JNJ组,JNJ组FAM96A、Bax表达水平高于培美曲塞组,联合组Bcl-2表达水平低于JNJ组,JNJ组Bcl-2表达水平低于培美曲塞组(均P<0.05)。联合组与单药组比较,血细胞计数均明显下降,肝功能指标均明显升高(P<0.05);培美曲塞组与JNJ组比较,血细胞及肝功能指标无统计学差异(P>0.05)。结论:JNJ-26481585可通过激活抑癌基因FAM96A的表达、上调Bax表达及下调Bcl-2的表达来促进癌细胞凋亡,逆转肺腺癌对培美曲塞的耐药性,并与培美曲塞协同发挥抗癌作用。

DNA甲基化在肺癌顺铂耐药中的研究进展

肺癌作为最常见的恶性肿瘤之一,严重威胁人类的生命健康。铂类药物如顺铂(cisplatin,DDP)是肺癌的一线治疗药物,但其获得性耐药是肺癌治疗的主要难题,也是预后不良的关键原因。顺铂耐药的发生机制较为复杂,确切机制尚未明确。大量研究表明DNA甲基化在肺癌顺铂耐药过程中发挥关键作用,DNA异常高甲基化使染色质构象发生变化,导致多种耐药基因和抑癌基因失活。阐明其复杂机制有望为寻找预测疗效的生物标志物和治疗新靶点的确定提供依据。因此,本文就DNA甲基化在肺癌顺铂耐药过程中的作用和机制做一综述,并总结近年来的去甲基化策略,为逆转肺癌顺铂耐药提供新思路。

MET基因改变的非小细胞肺癌靶向治疗耐药机制及治疗策略

间质表皮转化因子(mesenchymal to epithelial transition factor,MET)基因改变参与了非小细胞肺癌的增殖、侵袭和转移。MET-酪氨酸激酶抑制剂(tyrosine kinase inhibitors,TKIs)已获批用于MET基因改变的非小细胞肺癌,而这些药物的耐药不可避免。MET-TKIs的分子耐药机制错综复杂,尚不完全清楚。本文主要对这些MET-TKIs的潜在耐药机制进行综述,以期为MET基因改变的患者提供合理的治疗思路。

Co-transfection of MRP and bcl-2 antisense S-oligodeoxynucleotides reduces drug resistance in cisplatin-resistant lung cancer cells

Obejctive To detect the influence of antisense s oligodeoxynucleotides (S ODNs) of bd 2 and multidrug resistamce associated protein (MRP) genes multidrug resistance associated protein gene and bcl 2 antisense S oligodeoxynucleotides on cisplatin resistant lung adenocarcinoma cell line A 549 DDP which overexpresses both bcl 2 and MRP Methods A 549 DDP cells were treated with sense and antisense S ODN mediated by lipofection Expression of MRP and bcl 2 mRNA and protein in the treated cells was measured by RT PCR and flow cytometry (FCM), respectively Apoptosis was identified by DNA electrophoresis and terminal deoxynucleotidyl transferase (TdT) mediated biotin dUTP nick end labeling(TUNEL) The degree of drug resistance of the treated cells was detected by a cell viability 3' [4,5 dimethylthiazol 2 yl] 2,5 diphenyl tefrazolium bromide thiazolylblue (MTT) assay Results Expression of bcl 2 and MRP significantly decreased in the cells treated with bcl 2 or/and MRP antisense S ODN for 48h as compared to the cells untreated and sense treated ( P <0 05) Resistance to cisplatin in the cells treated with bcl 2 or/and MRP antisense S ODN decreased by 60 6% (6 5 times), 56 4% (7 2 times) and 71 0% (4 8 times), respectively, which paralleled the decrease of bcl 2 and MRP expression Similarly, the resistance to etoposide and epirubicin in antisense treated cells also reduced in parallel to decreases of the two gene expressions The drug resistance in sense treated cells was similar to that in untreated cells Statistically significant dose and concentration dependent increases of apoptotic cells were observed in the groups exposed to 100?μmol/L cisplatin for 48?h after treatment by bcl 2 or/and MRP antisense Conclusion Bcl 2 and MRP were at least additive and possibly synergistic in conferring drug resistance in a cisplatin resistant lung adenocarcinoma cell line Antisense S ODN could attenuate drug resistance by promoting cells apoptosis, which might lead to a new treatment for patients with non small cell lung cancers (NSCLCs) who are refractory to conventional chemotherapy

AKT激活与非小细胞肺腺癌吉非替尼耐药性的关系探究

目的 探讨丝氨酸-苏氨酸蛋白激酶B(AKT)激活与非小细胞肺腺癌吉非替尼耐药性的相关性。方法 选取2020年5月—2022年5月接受吉非替尼治疗的100例非小细胞肺腺癌为研究对象,依据是否发生吉非替尼耐药分为耐药组42例、非耐药组58例。比较2组癌组织中AKT蛋白表达情况。多因素Logistic回归分析吉非替尼耐药性的影响因素。比较耐药组不同临床病理特征患者AKT蛋白表达情况,分析其与临床病理特征相关性。受试者工作特征曲线评价不同影响因素对吉非替尼耐药性的预测效能。结果 耐药组癌组织中AKT蛋白表达水平高于非耐药组(P<0.05)。吸烟史、临床分期、淋巴结转移、AKT蛋白表达水平为非小细胞肺腺癌患者吉非替尼耐药的独立危险因素(P<0.01)。不同临床分期及淋巴结转移患者AKT蛋白表达水平比较差异有统计学意义(P<0.01)。耐药组AKT蛋白表达与临床分期、淋巴结转移呈正相关(P<0.01);AKT蛋白表达预测非小细胞肺腺癌患者吉非替尼耐药的曲线下面积(0.788)分别大于吸烟史(0.697)、临床分期(0.654)、淋巴结转移(0.640)预测(P<0.05,P<0.01)。结论 非小细胞肺腺癌吉非替尼耐药患者中AKT处于活化状态,且与临床分期、淋巴结转移密切相关,其对吉非替尼耐药性具有一定预测价值。

端粒酶催化亚基脱氧核酶抑制A549/DDP耐药细胞生长的实验研究

背景与目的端粒酶在多种肿瘤中均有表达,并且可能参与肿瘤耐药。本研究的目的是观察端粒酶催化亚基脱氧核酶对A549/DDP耐药细胞生长的抑制作用,探讨端粒酶作为耐药肿瘤细胞治疗新靶标的可能性。方法合成针对端粒酶催化亚基mRNA的脱氧核酶,脱氧核酶体外切割端粒酶催化亚基mRNA片段。采用TRAP法检测转染脱氧核酶的A549/DDP细胞端粒酶活性,采用MTT法分别检测脂质体转染脱氧核酶、顺铂及其混合物对A549/DDP细胞生长的作用。结果端粒酶脱氧核酶在体外可以切割端粒酶催化亚基RNA,并具有剂量依赖关系;转染脱氧核酶可抑制A549/DDP细胞端粒酶活性;端粒酶脱氧核酶0.25μmol/L作用72h,A549/DDP耐药细胞生长抑制率可达32.9%,同3mg/L顺铂联合使用时抑制率可达60.5%,两者相互作用指数CDI=0.9。结论端粒酶催化亚基脱氧核酶可明显降低端粒酶活性,显著抑制人肺腺癌A549/DDP耐药细胞的生长,并且与化疗药物顺铂有协同作用,提示端粒酶有可能成为耐药肿瘤细胞新的治疗靶标。

一条新的人肺腺癌耐药相关基因全长cDNA片段的克隆及序列分析

背景与目的:肺癌细胞的多药耐药(multi-drugresistance,MDR)是药物治疗失败的重要原因,但其机制尚未完全清楚。我们已经通过SSH发现了1条新的耐药相关基因片段,经检索GenBank,发现其与2001年4月公布的一条新的基因序列BC006151同源性高达99%。本研究旨在克隆该基因的全长,并检验该基因在几种肺癌细胞株中的表达,为进一步研究其功能和调控奠定基础。方法:根据BC006151基因序列设计引物,通过RT-PCR证实肺腺癌MDR细胞SPC-A-1/cDDP确实包含BC006151基因后,扩增该基因全长cDNA,通过测序对所得序列进行分析,并对其编码蛋白的氨基酸序列进行预测。应用半定量RT-PCR方法检验小细胞肺癌SH77、大细胞肺癌H460、肺腺癌SPC-A-1和SPC-A-1/cDDP等细胞株中该基因mRNA的表达。结果:成功克隆了BC006151基因的全长cDNA片段,长1298bp,它具有完整的阅读框和编码区,并与经SSH获得的片段有极高的同源性。该基因在MDR细胞株SPC-A-1/cDDP的表达明显高于其亲代细胞株;SPC-A-1表达高于H460,SH77未见表达。结论:BC006151基因是与cDDP致肺腺癌MDR密切相关的一条新基因。该基因全长cDNA片段的克隆将有助于阐明其在肺癌MDR发生中的作用。

人参皂苷Rg3对小鼠肺腺癌多耐药的逆转作用

目的观察人参皂苷Rg3对小鼠肺腺癌多耐药的逆转作用并探讨其机制。方法 将32只接种获得性多药耐药Lewis肺癌细胞的小鼠随机分成4组:荷瘤对照组、多柔比星(ADM)组、人参皂苷Rg3组和人参皂苷Rg3联合多柔比星(ADM)组。各组予以相应干预,观察肿瘤生长情况,肿瘤接种24天处死全部小鼠,剥取皮下移植肿瘤,称瘤重,计算抑瘤率,收集移植瘤标本行免疫组织化学检测P-糖蛋白(P-glycaprotein,p-Gp)和多药耐药相关蛋白1的表达,并行RT-PCR半定量检测多药耐药基因(multidrug resistance,MDR1)mRNA、多药耐药相关蛋白1(multidrug resistance-associated protein 1,MRP1)mRNA表达。结果 (1)各用药组肿瘤的生长受到不同程度抑制,瘤重低于对照组,其抑瘤率分别为10.5%、36.3%、56.2%,联合用药组抗瘤作用进一步增强。(2)免疫组织化学:p-Gp及MRPl蛋白质的表达具有一致性,对照组和ADM组表达较高,而人参皂苷Rg3组和联合组表达明显降低,其中对照组和ADM组比较,人参皂苷Rg3组和联合组比较,差异无统计学意义(P>0.05);而人参皂苷Rg3组和联合组与对照组和ADM组比较差异均有统计学意义(P<0.01)。(3)RT-PCR:与对照组和ADM组比较,人参皂苷Rg3组和联合组MDR1 mRNA荧光强度降低,人参皂苷Rg3组、联合组同对照组及ADM组比较MDR1 mRNA相对量减小,差异均有统计学意义(P<0.01),而对照组同ADM组比较,人参皂苷Rg3组同联合组比较差异未见统计学意义,MRP1 mRNA的表达与MDR1 mRNA类似。结论 人参皂苷Rg3对小鼠肺腺癌多药耐药有逆转作用,其作用机制可能是下调MDR1 mRNA、MRP1 mRNA及其蛋白的表达。

吉非替尼耐药细胞株的筛选和基因谱表达研究

目的:从对吉非替尼(gefitinib)敏感的肺腺癌细胞株PC9中筛选并建立了6株gefitinib耐药细胞亚克隆并研究其生物学特性。方法:采用N-甲基-N’-硝基-N-亚硝基胍(MNNG)诱变筛选和有限稀释法进行耐药性亚克隆的单细胞培养,MTT法检测肿瘤细胞对gefitinib的敏感性和IC50,DNA微列阵检测耐药细胞株与野生型PC9细胞的基因表达差异,免疫组织化学法测定细胞外间质成分fibronectin和collagenⅣ的表达差异,FCM法进行细胞周期分析。结果:获得了对gefitinib具有较高耐药性的2个细胞株:PC9/G2和PC9/G4,尤其PC9/G2的IC50为7～10μmol/L,较野生型PC9细胞(IC50:0.03～0.05μmol/L)提高耐受性160～260倍。PC9的细胞倍增时间为(16.2±3.5)h,PC9/G2的倍增时间为(36.5±4.8)h。PC9/G2同PC9的基因表达谱存在明显差异,耐药细胞株PC9/G2的脂肪酸代谢和氧化磷酸化基因表达下降,糖酵解基因表达上升,核糖体各亚基蛋白表达下调,高尔基体和囊泡、笼形蛋白衣被小泡等相关基因表达上调,细胞蛋白分泌功能加强;泛素-蛋白酶体循环相关基因上调;DNA修复基因和解旋酶类基因表达上调;具有蛋白激酶活性的30个基因表达上调,11个基因表达下调;15个磷酸化蛋白磷酸酶活性的基因表达上调;具有GTP结合活性的基因21个表达上调,4个下调;涉及细胞骨架和细胞运动功能的8个基因上调;整合素β1亚基、类胰岛素受体、NFκB级联反应的正向调节因子表达上调。结论:成功建立了6株PC9细胞的gefitinib耐药细胞株,初步研究表明对gefitinib中度耐药可能与细胞外基质组分改变及其黏附信号通路有关,对gefitinib高度耐药则可能涉及替代性信号通道的激活和信号通道的下游激活。

MiR-503逆转肺癌耐药细胞株A549/DDP的耐药性及其机制研究

背景与目的临床上肺癌细胞往往出现对顺铂的耐药性,因此探讨肿瘤细胞的耐药机制,开发新的逆转耐药性的方法,对提高临床患者的受益有十分重要的意义。miRNA可通过其调控的目标基因,对多种与肿瘤细胞失控生长、抗凋亡、迁移和侵袭,甚至是肿瘤细胞对药物治疗的应答产生调控作用。本实验旨在探讨miR-503对肺癌顺铂耐药细胞株A549/DDP的耐药性逆转及其相关作用机制。方法应用MTS法检测miR-503对A549/DDP细胞顺铂耐受性的影响,流式细胞术检测肿瘤细胞凋亡率以及胞内罗丹明-23(Rhodamine-23,Rh-23)含量的变化,Western blot法和Real time PCR检测肿瘤细胞多药耐药蛋白MDR、MRP、Survivin和Bcl-2蛋白表达,以及Akt磷酸化的变化,应用双萤光报告基因技术检测细胞NF-κB和AP-转录活性。结果与对照细胞组相比较,miR-503转染A549/DDP细胞株后,可明显增加细胞对顺铂的敏感性,使耐药逆转倍数增加为2.48倍,Rh-23含量升高2.49倍,细胞凋亡率提高0.3倍;在转录水平检测发现,与对照组相比较,miR-503转染的细胞中MDR、MRP、ERCC、Survivin及Bcl-2等与肿瘤耐药相关基因的mRNA表达水平明显下调,而RhoE mRNA表达水平则明显升高(P<0.05);进一步在蛋白水平亦证实MDR、MRP、ERCC、Survivin、Bcl-2以及p-Akt的表达明显下降,RhoE的表达明显上升。结论 miR-503可逆转A549/DDP对顺铂的耐药性,这一作用可能与抑制药物外排,负调控肿瘤耐药相关蛋白的表达,促进细胞凋亡有关。

淫羊藿苷逆转耐甲氨蝶呤肺癌A549细胞转移表型

目的:研究中药淫羊藿苷(icariin,ICA)作用甲氨蝶呤(methotrexate,MTX)耐药肺癌A549细胞后对细胞转移表型的影响,初步探讨ICA逆转A549/MTX耐药细胞转移表型的作用机制及对肺癌的治疗价值。方法:采用MTT法检测ICA对A549/MTX耐药细胞的半数抑制浓度(half inhibition concentration,IC50)。采用双层软琼脂克隆形成实验检测A549/MTX组和A549/MTX+ICA组细胞的克隆形成率,并观察其集落形态。细胞划痕实验检测A549/MTX组和A549/MTX+ICA组细胞的迁移能力。Transwell小室实验检测细胞侵袭能力的变化。结果:MTT结果显示,无毒剂量的ICA与MTX联合应用后A549/MTX细胞的IC50值为35.50±1.85μmol/L,比单独应用MTX(同等剂量)后A549/MTX细胞的IC50值(52.17±2.25μmol/L)有了一定程度的下降。软琼脂实验发现,A549/MTX+ICA组细胞克隆形成率为0.94±0.09,小于A549/MTX组细胞的1.56±1.07(P<0.05)。划痕实验显示,72 h后A549/MTX组细胞的迁移能力大于A549/MTX+ICA组细胞(P<0.05)。Transwell实验显示,A549/MTX组细胞的穿膜细胞数明显多于A549/MTX+ICA组细胞(P<0.05),说明A549/MTX+ICA组细胞的侵袭浸润能力小于A549/MTX组细胞。结论:中药ICA具有逆转A549/MTX耐药细胞转移表型的作用。

EGFR和LRP表达与卵巢癌化疗耐药及预后的关系

背景与目的:表皮生长因子受体(epidermal growth factor receptor,EGFR)的异常表达和活化能引起肿瘤细胞化疗耐药的产生,与卵巢癌化疗后复发及预后密切相关。肺耐药蛋白(lung resistance protein,LRP)是一种主要介导铂类等化疗药物耐药的多药耐药蛋白。研究表明,LRP是预测卵巢癌化疗敏感的独立预后因素。本研究探讨EGFR和LRP表达与卵巢癌化疗耐药及预后的关系。方法:采用免疫组化PV-6000二步法,检测76例恶性卵巢肿瘤、9例卵巢交界性肿瘤、17例卵巢良性肿瘤和15例卵巢正常组织中EGFR和LRP的表达,分析EGFR和LRP表达与卵巢癌化疗疗效及患者术后生存时间的关系。结果:卵巢癌组织中EGFR和LRP的阳性率分别为73.68%和71.79%,均显著高于正常卵巢组织和良性肿瘤组织(P<0.01);EGFR高表达于Ⅲ～Ⅳ期、低分化和有腹水的卵巢癌组织中(P<0.05)。EGFR和LRP阳性者近期化疗有效率分别为57.14%和53.70%,低于阴性者(P<0.05);化疗耐药型卵巢癌患者EGFR和LRP阳性率分别为92.86%和85.71%,高于化疗敏感型(P<0.05)。生存分析表明,卵巢癌患者3年生存率为53.00%。EGFR、LRP阳性和近期化疗疗效无效者术后生存时间短(P<0.01)。结论:EGFR和LRP可作为预测卵巢癌化疗耐药及预后的指标。

晚期非小细胞肺癌ERCC1和BRCA1的表达及与顺铂耐药性的临床研究

目的:探讨核苷酸切除修复交叉互补组1(excision repair cross complementation group 1,ERCC1)和乳腺癌易感基因(breast cancer susceptibility gene 1,BRCA1)在非小细胞肺癌(non-small cell lung cancer,NSCLC)中的表达及与顺铂耐药的关系。方法:应用免疫组织化学法检测81例NSCLC患者标本ERCC1和BRCA1蛋白的表达,并与患者年龄、性别、组织病理学类型、临床分期、吸烟指数及含铂化疗方案疗效的关系进行分析。结果:ERCC1蛋白表达阳性率为35.80%,与患者的性别、年龄、临床分期及吸烟指数无关,ERCC1蛋白在腺癌中的表达高于鳞癌,差异有统计学意义(P<0.05);ERCC1表达阳性组的化疗有效率(24.14%)低于阴性组(63.46%),差异有统计学意义(P<0.05);ERCC1表达阳性组的中位生存期(median sur-vival time,MST)(12.1个月)低于阴性组(20.6个月),差异有统计学意义(P<0.05)。BRCA1蛋白表达阳性率为51.85%,与患者的性别、年龄、临床分期、组织学类型及吸烟指数无关;BRCA1表达阳性组化疗有效率(38.10%)低于阴性组(61.54%),差异有统计学意义(P<0.05);BRCA1表达阳性组的MST(15.2个月)与阴性组(16.7个月)比较无明显差异。结论:ERCC1和BRCA1蛋白表达检测可预测NSCLC患者对铂类化疗药物的敏感性,有助于临床上NSCLC个体化化疗方案的制定。

薏苡仁注射液对肺腺癌细胞多药耐药性逆转作用的机制研究

目的观察薏苡仁注射液(康莱特)对人肺腺癌细胞A549多药耐药性(MDR)逆转作用,并对其机制进行探讨。方法体外培养人肺腺癌细胞系A549并以不同剂量康莱特进行处理,磺酰罗丹明(sulphorhodamine B,SRB)染色法检测药物对各组细胞的抑制率;反转录-聚合酶链反应法(RT-PCR)检测多药耐药因子多药耐药基因1(MDR1)、生存素(Survivin)、B细胞淋巴瘤/白血病-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)mRNA在各组中表达情况;蛋白质印迹(Western blot)法检测P-糖蛋白(P-gp)、Survivin、Bcl-2、Bax蛋白水平表达,将各组结果进行比较。结果 4组肺腺癌细胞A549的抑制率比较,差异有统计学意义(P<0.05)。其中,与空白对照组比较,康莱特低、中、高剂组抑制率降低,差异均有统计学意义(P<0.05)。康莱特中、高剂组抑制率比较,差异亦有统计学意义(P<0.05)。康莱特低、中、高剂组MDR1、P-gp、Survivin、Bcl-2表达降低,Bax表达升高,差异均有统计学意义(P<0.05)。结论康莱特可抑制肺腺癌细胞生长,该药具有明显的逆转肺腺癌多药耐药性的作用,该作用是通过调节多药耐药因子的表达而实现的。

肺癌组织中突变型p53蛋白与肿瘤耐药蛋白表达对治疗及预后影响的研究

目的:探讨突变型p53蛋白与肿瘤耐药蛋白在肺癌组织的表达特点及其临床意义。方法:免疫组化法检测66例肺癌组织中突变型p53蛋白与各种肿瘤耐药蛋白的表达,并分析治疗及其效果和生存情况。结果:不同TNM分期(χ2=6.22,P=0.016)、淋巴结转移(χ2=3.88,P=0.045)、化疗效果(χ2=4.78,P=0.035)及3年生存组(χ2=5.10,P=0.024)的突变型p53蛋白表达均差异有统计学意义;而P-糖蛋白(P-gp)、多药耐药相关蛋白(MRP)、肺耐药相关蛋白(LRP)和谷胱甘肽-S-转移酶-π(GST-π)的表达仅对近期化疗效果产生影响,P值分别为0.035、0.018、0.041和0.021。仅突变型p53蛋白阳性组和阴性组的3年生存率、中位生存期差异有统计学意义,χ2=5.137,P=0.023;不同病理类型肺癌患者3年生存率、生存时间的差异也有统计学意义,χ2=19.5,P=0.001。结论:突变型p53蛋白的检测能在一定程度上反映肺癌的TNM分期、淋巴结转移、化疗效果和预后;而P-gp、MRP、GST-π和LRP的检测只能提示肺癌的近期化疗效果;不同病理类型的肺癌患者预后不同。

贝伐单抗在肺癌中的研究进展

肺癌是目前最常见的恶性肿瘤之一,其发病率有逐年增高的趋势,且患者的预后及生存期很差。贝伐单抗于2004-02-26获得美国食品药品监督管理局(FDA)批准,是美国第一个批准上市的抑制肿瘤血管生成的药物,在肺癌的药物治疗中占据重要地位。目前贝伐单抗在肺癌中的研究多从1或2个角度来论述,难以让读者清楚贝伐单抗与肺癌的关系。本文主要从血管内皮生长因子在肿瘤发病机制中的作用、贝伐单抗治疗肺癌的机制、所检测的生物标志物、分期研究、耐药机制及毒副作用等角度较为全面地阐述贝伐单抗在肺癌中的研究进展。