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.     

Inhibiting Pathways Predicted From a Steroid Hormone Gene Signature Yields Synergistic Antitumor Effects in NSCLC

IntroductionMounting evidence supports a role for estrogen signaling in NSCLC progression. We previously reported a seven-gene signature that predicts prognosis in estrogen receptor β positive (ERβ+) NSCLC. The signature defines a network comprised of ER and human EGFR-2/3 (HER2/HER3) signaling.MethodsWe tested the efficacy of combining the pan-HER inhibitor, dacomitinib, with the estrogen antagonist, fulvestrant, in ERβ+ NSCLC models with differing genotypes. We assessed the potency of this combination on xenograft growth and survival of host mice, and the ability to reverse the gene signature associated with poor outcome.ResultsSynergy was observed between dacomitinib and fulvestrant in three human ERβ+ NSCLC models: 201T (wild-type EGFR), A549 (KRAS mutant), and HCC827 (EGFR 19 deletion) with combination indices of 0.1-0.6. The combination, but not single agents, completely reversed the gene signature associated with poor prognosis in a mechanism that is largely mediated by activator protein 1 downregulation. In vivo, the combination also induced tumor regression and reversed the gene signature. In HCC827 xenografts treated with the combination, survival of mice was prolonged after therapy discontinuation, tumors that recurred were less aggressive, and two mechanisms of HER inhibitor resistance involving c-Met activation and PTEN loss were blocked.ConclusionsThe combination of an ER blocker and a pan-HER inhibitor provides synergistic efficacy in different models of ERβ+ NSCLC. Our data support the use of this combination clinically, considering its ability to induce potent antitumor effects and produce a gene signature that predicts better clinical outcomes.     

Occludin在非小细胞肺癌组织中的表达及其临床意义

目的:检测非小细胞肺癌（NSCLC）组织中闭合蛋白（Occludin）的表达水平及其临床意义。方法:应用免疫组织化学方法检测73例NSCLC组织及其配对正常肺组织中Occludin蛋白表达水平,分析Occludin蛋白的表达差异及其与临床病理参数的相关性。结果:Occludin蛋白在肿瘤组织中的阳性表达率显著低于配对正常肺组织（P＜0.05）。Occludin蛋白的表达水平与患者性别、肿瘤大小、淋巴结转移、组织学类型、Ki-67表达水平显著相关。结论:Occludin蛋白表达下调与NSCLC细胞的增殖、转移能力相关,在肿瘤进展中发挥重要作用。     

miR-338通过靶向GPX4抑制非小细胞肺癌细胞增殖

目的:探讨miRNA-338（miR-338）通过靶向调控谷胱甘肽过氧化物酶4（GPX4）表达在非小细胞肺癌(non-small cell lung cancer,NSCLC)增殖中的作用及机制研究。方法:通过实时定量聚合酶链式反应（qRT-PCR）检测非小细胞肺癌组织、癌旁、细胞系及对照细胞系中miR-338及GPX4 mRNA的表达水平;通过Western Blot检测非小细胞肺癌组织、癌旁、细胞系及对照细胞系中GPX4蛋白水平;通过荧光素酶报告基因时间验证miR-338直接靶向调节GPX4的表达;通过CCK-8探索miR-338是否通过调节铁死亡影响肿瘤细胞增殖;通过试剂盒检测细胞脂质氧化和活性氧水平。结果:NSCLC组织和细胞系中miR-338表达水平低于癌旁组织和对照细胞系;NSCLC组织和细胞系中GPX4 mRNA及蛋白表达水平高于癌旁组织和对照细胞系;Starbase软件分析发现GPX4 mRNA序列中含有miR-338特异作用位点,荧光素酶报告基因实验结果证实miR-338直接靶向调节GPX4表达;过表达miR-338提高肿瘤细胞中脂质氧化及活性氧水平,并抑制肿瘤细胞增殖;而铁死亡抑制剂预处理可以逆转miR-338的抑癌作用。结论:miR-338通过负向调控GPX4表达进而促进肿瘤细胞铁死亡,最终抑制NSCLC细胞增殖。     

Efficacy and safety of anlotinib plus S-1 as thirdly-line or later-line treatment in advanced non-small cell lung cancer

Objective Anlotinib,an oral vascular endothelial growth factor receptor 2(VEGFR2)inhibitor,has confirmed antitumor activity in lung cancer in both in vitro and in vivo assays,and has been recommended as third-line treatment agent in non-oncogene driven non-small cell lung cancer(NSCLC).This prospective study aimed to investigate the efficacy and safety of anlotinib plus S-1 for third-or later-line treatment in patients with advanced NSCLC.Methods Patients with histologically or cytologically confirmed NSCLC,and documented disease progression following second-line chemotherapy,and/or epidermal growth factor receptor-tyrosine kinase inhibitor(EGFR-TKI)treatment were enrolled in this study.The patients were treated anlotinib(8 mg daily d 1–14)and S-1(60 mg/m^2 d 1–14)and the treatment was repeated every 3 weeks.Treatment was continued until disease progression or unacceptable toxicity occurred.The objective response rate(ORR),disease control rate(DCR),progression-free survival(PFS),and adverse events(AEs)were reviewed and evaluated.Results Forty-one patients were enrolled in the study between June 2018 and December 2018.The total ORR and DCR were 26.8%and 80.5%,respectively.The median PFS was 5.2 months[95%confidence interval(CI),3.9 to 6.6 months].In the univariate analysis,there was a significant difference in the median PFS between patients with brain metastases and those without brain metastases(4.8 months vs 5.9 months,respectively;P=0.039).The Eastern Cooperative Oncology Group(ECOG)performance status(P=0.002),lines of therapy(P=0.015),and therapeutic evaluation(P=0.014)were independent factors that influenced PFS.The most common AEs were hypertension,proteinuria,myelosuppression,gastrointestinal reactions,fatigue,and mucositis.Conclusion Anlotinib plus S-1 is an effective and safe regimen for advanced NSCLC as third-or later-line therapy.     

Expression and prognostic significance of m6A‐related genes in TP53‐mutant non‐small‐cell lung cancer

BackgroundTP53 is an important tumor suppressor gene on human 17th chromosome with its mutations more than 60% in tumor cells. Lung cancer is the highest incidence malignancy in men around the world. N‐6 methylase (m6A) is an enzyme that plays an important role in mRNA splicing, translation, and stabilization. However, its role in TP53‐mutant non‐small‐cell lung cancer (NSCLC) remains unknown.MethodFirst, we investigated 17 common m6A regulators'' prognostic values in NSCLC. Then, after the establishment of risk signature, we explored the diagnostic value of m6A in TP53‐mutant NSCLC. Finally, gene set enrichment analysis (GSEA), gene ontology (GO) enrichment analysis, and differential expression analysis were used to reveal the possible mechanism of m6A regulators affecting TP53‐mutant NSCLC patients.ResultsStudy showed that nine m6A regulators (YTHDC2, METTL14, FTO, METTL16, YTHDF1, HNRNPA2B1, RBM15, KIAA1429, and WTAP) were expressed differently between TP53‐mutant and wild‐type NSCLC (p < 0.05); and ALKBH5 and HNRNPA2B1 were associated with the prognostic of TP53‐mutant patients. After construction of the risk signature combined ALKBH5 and HNRNPA2B1, we divided patients with TP53 mutations into high‐ and low‐risk groups, and there was a significant survival difference between two groups. Finally, 338 differentially expression genes (DEGs) were found between high‐ and low‐risk groups. GO enrichment analysis, PPI network, and GSEA enrichment analysis showed that m6A may affect the immune environment in extracellular and change the stability of mRNA.ConclusionIn conclusion, m6A regulators can be used as prognostic predictors in TP53‐mutant patients.     

Hsa_circRNA_0017620 regulated cell progression of non‐small‐cell lung cancer via miR‐520a‐5p/KRT5 axis

BackgroundCircRNA is a very important functional RNA that plays an important role in the development and metabolism of cancer. However, the study of circRNA in NSCLC has not been fully elucidated.MethodsThe expression of hsa_circ_0017620, SFMBT2, miR‐520a‐5p, and KRT5 was determined using qRT‐PCR. KRT5, Twist1, E‐cadherin, and Ki67 protein expression were measured with western blot. The positive expression rates of Ki67 and Vimentin were determined by immunohistochemistry assay. 5‐Ethynyl‐2’‐deoxyuridine (EdU), colony formation, and MTT assays were used to assess cell proliferation. Transwell migration and invasion assay were applied to determine cell migration and invasion. Dual‐luciferase reporter and RNA immunoprecipitation assays were used to verify the relationship among hsa_circ_0017620, miR‐520a‐5p, and KRT5. The animal experiment was used to ensure the effects of hsa_circ_0017620 on tumor growth in vivo.ResultsHsa_circ_0017620 was upregulated in NSCLC cells and tissues. MiR‐520a‐5p had been verified to be a target miRNA of hsa_circ_0017620 and KRT5 had been verified to be a target mRNA of miR‐520a‐5p in NSCLC cells. Knockdown of hsa_circ_0017620 inhibited cell proliferation, migration, and invasion in NSCLC cells, which was reversed by downregulating miR‐520a‐5p or upregulating KRT5 in NSCLC. Overexpression of hsa_circ_0017620 had opposite effects in NSCLC. Moreover, hsa_circ_0017620 silencing inhibited tumor growth in vivo of NSCLC.ConclusionIn this study, we found that hsa_circ_0017620 played an important role in NSCLC progression. Hsa_circ_0017620 regulated cell proliferation, invasion, and migration through targeting miR‐520a‐5p/KRT5 axis in NSCLC, providing a potential new target for the treatment and diagnosis of NSCLC.     

非小细胞肺癌中人表皮生长因子受体-2 蛋白的表达及临床意义

目的 检测非小细胞肺癌(NSCLC)中HER-2/neu蛋白的表达及其临床病理意义,探讨应用HER-2/neu单抗治疗NSCLC的可能性.方法 应用DAKO HercepTest试剂盒,对105例外科切除、病理诊断为NSCLC标本的HER-2/neu蛋白的表达情况进行检测,并对其与临床病理特征的关系进行统计学分析.结果 105例NSCLC中,有80例(76%)出现HER-2/neu蛋白表达,其中41例(39%)大于10%的区域有阳性表达,胞膜显色不连续;24例(23%)大于10%的区域有阳性表达,胞膜显色连续,强度中等;15例(14%)大于10%的区域有阳性表达,胞膜呈连续强阳性表达;组织HER-2过度表达率为37%(39/105).HER-2蛋白过度表达与NSCLC的病理类型无密切关系;过度表达的肺癌淋巴结转移率显著高于非过度表达者(P＜0.05);不同的肺癌TNM分期具有不同的HER-2过度表达率,后者随TNM分期的增加而显著增加(P＜0.05);肺癌HER-2过度表达的术后2年生存率显著低于非过度表达者(P＜0.05).结论 HER-2/neu蛋白是NSCLC的一个重要预后指标,HER-2/neu阳性病例有可能应用抗HER-2/neu单克隆抗体Herceptin进行靶向治疗.     

洛铂联合吉西他滨治疗晚期肺癌安全性的临床观察

目的观察评价新一代铂类抗癌药物洛铂（labaplatin,LBP）联合吉西他滨（gemicitabine,GEM）组成的GP方案治疗晚期肺腺癌和晚期小细胞肺癌（SCLC）的安全性。方法开放性、单试验组、Ⅰ期临床研究,共入组2例,均为常规放化疗效果差的患者,男性1例,女性1例,年龄分别为79岁、53岁。病理类型分别为小细胞肺癌（广泛期）、肺腺癌（Ⅳ期）。应用GP方案,即LBP30mg／m^2,静滴,d1;GEM1000mg／m^2,静滴,d1.8,21d为1个周期。其中男性SCLC患者接受1个周期化疗、女性NSCLC患者接受1．5个周期化疗,按照WHO和NCI标准评价客观疗效和毒副反应,定期随访。结果2例毒副反应主要表现为可逆性的骨髓抑制、胃肠道反应、一过性肝损害、脱发,未见明显心肾毒性。结论LBP联合GEM组成GP方案治疗晚期NSCLC和SCLC的毒副反应可以耐受,可进一步研究观察。