MicroRNA-145增强非小细胞肺癌细胞对吉非替尼敏感性及其机制探讨

目的:探讨微小RNA-145（microRNA-145，miR-145）对非小细胞肺癌细胞系吉非替尼耐药的作用及其可能的潜在机制。方法:实时荧光定量PCR（Q-PCR）方法检测正常细胞及非小细胞肺癌细胞中miR-145的表达差异；不同时间5 μmol/L吉非替尼干预肺癌细胞SPC-A-1和A549后，Q-PCR检测miR-145表达变化；miR-145 mimics和miR-145 inhibitors分别转染SPC-A-1和A549肺癌细胞后，CCK8检测细胞活力变化;双荧光素酶报告系统检测miR-145与ADAM19的靶向结合；miR-145 mimics转染SPC-A-1和A549肺癌细胞，Western blot检测ADAM19蛋白表达；Western blot检测5 μmol/L吉非替尼干预后，SPC-A-1和A549肺癌细胞中ADAM19蛋白的表达；miR-145 mimics转染SPC-A-1和A549肺癌细胞，再用5 μmol/L吉非替尼干预，Western blot检测ADAM19蛋白的表达；采用siRNA抑制ADAM19表达，再用5 μmol/L吉非替尼干预，CCK8检测细胞活力；采用BALB/C雌性裸鼠皮下接种肿瘤细胞的方法，观察上述效应。结果:Q-PCR结果显示，与正常肺上皮细胞系BEAS-2B相比较，肺癌细胞SPC-A-1和A549中miR-145表达明显降低；5 μmol/L吉非替尼干预SPC-A-1和A549细胞后，miR-145表达显著上调；转染miR-145 mimics后,CCK8结果显示两种细胞细胞活力下降；双荧光素酶报告系统结果显示，miR-145靶向结合ADAM19基因的3'-UTR区；Western blot结果显示，5 μmol/L吉非替尼诱导SPC-A-1和 A549细胞后，两种细胞中ADAM19蛋白表达均降低；miR-145 mimics转染SPC-A-1和A549细胞，之后给予5 μmol/L吉非替尼治疗，ADAM19蛋白表达下调；siRNA抑制SPC-A-1和A549细胞中ADAM19表达，再给予5 μmol/L吉非替尼治疗，CCK8结果显示，两种细胞的细胞活力显著降低；过表达BALB/C雌性裸鼠的miR-145后，显著抑制皮下肿瘤生长，并且增强吉非替尼的抗肿瘤效果。结论:miR-145显著增强肺癌细胞SPC-A-1和A549对吉非替尼的敏感性，其机制可能是通过靶向结合AMDM19基因的3'-UTR区域，从而抑制其表达。miR-145有可能成为治疗非小细胞肺癌吉非替尼耐药的有效靶点。     

吉非替尼联合培美曲塞和顺铂治疗晚期非小细胞肺癌的安全性和有效性

目的:比较单用培美曲塞/顺铂化疗或吉非替尼联合培美曲塞/顺铂化疗治疗晚期非小细胞肺癌（non-small cell lung cancer，NSCLC）的临床疗效及安全性，为临床应用提供参考。方法:选择标准一线化疗后复发的晚期NSCLC患者112例，其中联合治疗组52例接受培美曲塞/顺铂及吉非替尼治疗，化疗组60例仅采用培美曲塞和顺铂化疗，评价两组患者的临床疗效及不良反应。结果:联合组客观有效率（objective response rate，ORR）为36.5%，高于化疗组的18.3%（P=0.030）；两组疾病控制率（disease control rate，DCR）分别为71.2%和55.0%（P=0.035）；两组患者的中位无进展生存期（progression free survival，PFS）分别为8.7个月和6.7个月，差别有统计学意义（P=0.047），但两组患者的中位总生存期（overall survival，OS）差别无统计学意义（P=0.265）。与治疗前相比，两组患者的肿瘤标志物均明显下降，但联合组的CEA和CYFRA21-1水平比化疗组更低（P<0.05）。联合组皮疹和腹泻的发生率高于化疗组（P<0.05），两组之间其它不良反应的发生率无明显差别（P>0.05）。结论:晚期NSCLC患者一线化疗失败后，采用培美曲塞/顺铂化疗联合吉非替尼靶向治疗较单用化疗显示出更高的ORR和中位PFS，且不良反应可以耐受，值得临床推广运用。     

A Randomized Phase 2 Study of Gefitinib With or Without Pemetrexed as First-line Treatment in Nonsquamous NSCLC With EGFR Mutation: Final Overall Survival and Biomarker Analysis

IntroductionClinical studies have shown that a combination of a tyrosine kinase inhibitor (TKI) and pemetrexed overcame acquired resistance to epidermal growth factor receptor (EGFR) TKI in NSCLC. Previously, pemetrexed+gefintib (P+G) had improved progression-free survival (PFS) compared with gefitinib. We present OS, updated PFS, biomarker analysis, and safety of P+G versus gefitinib.MethodsThis was a phase 2, multicenter, randomized study conducted in East Asian patients with advanced nonsquamous NSCLC with EGFR mutations. Patients were randomized (2:1) to receive P+G (500 mg/m² intravenously 3-weekly + 250 mg/day orally) or gefitinib.ResultsIn total, 191 patients (P+G, n=126; gefitinib, n=65) comprised the intent-to-treat and safety populations. Median OS was 43.4 months in P+G versus 36.8 months in gefitinib arm; adjusted HR 0.77 (95% CI, 0.5-1.2); one-sided P=0.105. Median PFS was significantly longer in the P+G (16.2 months) versus gefitinib arm (11.1 months); adjusted HR 0.67 (95% CI, 0.5-0.9); one-sided P=0.009. In the P+G and gefitinib arms, median PFS was 22.6 and 11.0 months, respectively, in patients with low thymidylate synthase (TS) expression, and 12.6 and 9.9 months, respectively, in patients with high TS expression. Common second-line post-discontinuation systemic therapies were EGFR-TKIs and chemotherapy. Most patients experienced at least one adverse event.ConclusionsAddition of pemetrexed to EGFR TKI gefitinib resulted in significantly improved PFS and numerically longer OS compared with gefitinib in treatment-naïve patients with EGFR-mutated advanced nonsquamous NSCLC. Low TS expression appeared to be a good predictor for treatment outcomes.     

肺癌靶向治疗研究:以文献计量学和可视化分析描述的热点与趋势

目的:通过文献计量学分析方法，阐述并分析肺癌靶向治疗的研究热点与趋势。方法:文章检索了万方数据库、Web of Science、SooPAT(中国专利)数据库近10年来国内外肺癌靶向治疗的相关文献，以文献计量学方法，分析并归类了肺癌靶向治疗的研究热点与时效变迁的趋势。结果:在万方数据库3 744篇肺癌靶向治疗的中文文章中，发现作用于表皮生长因子受体基因突变位点的分子靶向药物-吉非替尼治疗非小细胞肺癌的研究是中国学者近10年的主要研究热点；在近10年Web of Science数据库中检索到的810篇英文文章及参考文献显示，西妥昔单抗、贝伐单抗、纳武单抗、吉非替尼、厄洛替尼和环唑替尼为这10年肺癌治疗领域靶向治疗的热点药物，而多种靶向药物的联合治疗、晚期非小细胞肺癌的靶向治疗效果和肺癌靶向治疗后的脑转移也成为近5年该领域的国际关注热点；中国肺癌靶向治疗领域的254项有权专利的分析显示，申报了专利的主要药物包括酸敏感的吉非替尼-氟硼二吡咯衍生物、顺铂抗肺癌主动靶向隐形类脂质体等。结论:文章采用文献计量学的量化分析技术，呈现出近10年肺癌靶向治疗已成为该领域专业研究者持续关注的热点，而如何选择正确的靶向药物或药物组合方案来解决肺癌治疗中的耐药性问题，同时最大程度地减少患者的不良反应，以延长患者的生存率，应是未来集中研究的方向。     

Interactions of crizotinib and gefitinib with organic anion-transporting polypeptides (OATP)1B1, OATP1B3 and OATP2B1: gefitinib shows contradictory interaction with OATP1B3

1.?The drug–drug interaction (DDI) mediated by organic anion-transporting polypeptide (OATP)1B1, OATP1B3 and OATP2B1 has a major impact on the hepatic clearance of drugs. The effects of tyrosine kinase inhibitors (TKIs) on OATPs have not been well studied. In the present study, we evaluated the contribution of OATPs to the hepatic uptake of crizotinib and gefitinib and the interaction of those TKIs with OATPs to estimate DDIs.

2.?To clarify whether crizotinib and gefitinib were substrates for OATPs, we performed uptake studies. We examined the effects of the TKIs on uptake of typical substrates and fluvastatin via OATPs. IC₅₀ and EC₅₀ values of the TKIs were calculated.

3.?OATP1B3- and OATP2B1-mediated crizotinib uptake and OATP2B1-mediated gefitinib uptake were observed. Gefitinib accelerated OATP1B3-mediated [³H]TCA uptake and inhibited OATP2B1-mediated [³H]E₃S uptake. On the other hand, gefitinib inhibited OATP1B1- and OATP2B1-mediated fluvastatin uptake.

4.?We provided basic information to estimate the DDI on OATPs caused by TKIs. The DDI on OATPs caused by gefitinib could occur in a normal clinical situation. And the uptake of crizotinib into the intrahepatocellular environment via OATPs may induce DDI and liver damage. We therefore emphasize the necessity of careful use of TKIs.     

抗瘤增效方联合吉非替尼治疗老年非小细胞肺癌患者及对炎症因子,T细胞亚群水平及血清肿瘤标志物的影响

目的:探讨抗瘤增效方联合吉非替尼治疗老年非小细胞肺癌及对患者炎症因子,T细胞亚群水平及血清肿瘤标志物影响。方法:选取郑州大学附属肿瘤医院肿瘤内科非小细胞肺癌Ⅳ期老年患者84例,患者或家属签字同意,积极配合此次研究,按随机数字表法分组,对照组患者(42例)予以单纯吉非替尼治疗,研究组患者(42例)予以吉非替尼联合抗瘤增效方治疗,观察并记录所有患者治疗前后生存质量、炎症因子及T细胞亚群水平,同时对比临床疗效及不良反应状况。结果:对照组有效率低于研究组(P0.05);与治疗前比较,两组患者治疗后生存质量、炎症因子及T细胞亚群水平均发生变化,研究组治疗后躯体功能,角色功能,社会功能,情绪功能评分高于对照组,研究组治疗后白细胞介素-2(interleukin-2,IL-2),白细胞介素-12(interleukin-12,IL-12)和γ-干扰素(interferon-γ,IFN-γ)水平高于对照组,研究组治疗后CD8+水平低于对照组,CD4+及CD4+/CD8+水平高于对照组(P0.05);治疗后,对照组血管内皮生长因子(vascular endothelial growth factor,VEGF)升高,治疗组VEGF下降(P0.05);与治疗前比较,两组患者癌胚抗原(carcinoembryonic antigen,CEA),糖抗原(carbohydrate antigen-125,CA125)及细胞角蛋白19片段21-1(cytokeratin 19 fragments,CYFRA21-1)差异无统计学意义,组间比较差异无统计学意义;两组患者不良反应较轻,无药物不良反应。结论:抗瘤增效方联合吉非替尼治疗老年非小细胞肺癌疗效确切,能提高生活质量及免疫功能。     

Profile of the therascreen® EGFR RGQ PCR kit as a companion diagnostic for gefitinib in non-small cell lung cancer

Introduction: Gefitinib is recently approved by the US Food and Drug Administration as a first-line treatment for non-small cell lung cancer (NSCLC) patients harboring EGFR mutations. The therascreen® EGFR RGQ PCR Kit is approved as a companion diagnostic to select patients with EGFR exon 19 deletions and L858R mutation for treatment with gefitinib.

Areas covered: This article reviews the methods for detecting EGFR mutations, the technology and indication of the therascreen® kit, and the clinical utility of the assay in phase 3 and phase 4 clinical trials. Studies that compared the performance of the therascreen® kit with other assays and assessed the kit’s application in non-tissue samples are also discussed.

Expert commentary: The therascreen® kit is a highly sensitive real-time polymerase chain reaction test that provides standardised testing and automated interpretation of EGFR mutation status in formalin-fixed, paraffin-embedded (FFPE) tissue samples. Although not indicated for these applications, the test has also shown utility in detecting uncommon sensitizing EGFR mutations as well as mutations in non-tissue samples.