自噬通路基因多态性与晚期非小细胞肺癌含铂化疗疗效的相关性分析

对晚期非小细胞肺癌患者采用含铂化疗是肺癌临床治疗中非常重要的方法,然而不同患者对含铂化疗的敏感性却存在着明显的个体差异,这提示发现潜在的分子标志物对预测临床中含铂化疗疗效具有关键作用。本研究旨在探索自噬通路基因多态性与晚期非小细胞肺癌含铂化疗疗效之间的相关性,以期寻找可能影响含铂化疗药物敏感性的分子标记。本研究纳入了1004例接受含铂化疗的晚期非小细胞肺癌患者,分析了自噬通路中13个基因上的99个SNP位点与含铂化疗临床获益、无疾病进展时间及总生存时间之间的相关性。研究发现,位于ULK1基因的位点rs7953348(G>A) (P=0.017, OR:0.67, 95%CI:0.49~0.93)和rs12303764(A>C) (P=0.009, OR:0.63, 95%CI:0.45-0.89)及位于ATG14基因上的位点rs17742719(C>A) (P=0.002, OR:1.83, 95%CI:1.26~2.66)、rs8003279(A>G) (P=0.006, OR:1.65, 95%CI:1.16~2.35)和rs1009647(G>A) (P=0.002, OR:1.70, 95%CI:1.22~2.37)与临床获益存在显著关联,位于DRAM基因上的位点rs7955890(G>A) (P=0.004, HR:0.63; 95%CI:0.46~0.86)和rs17032060(G>A) (P=0.006, HR:0.65, 95%CI:0.48~0.88)及位于ATG3基因上的位点rs13082005(G>A) (P=0.012, HR:1.27,95%CI:1.05~1.53)与含铂化疗的无疾病进展时间显著相关,位于ULK1基因的位点rs7953348(G>A) (P=0.011, HR:0.74, 95%CI:0.58~0.93)和位于ATG10基因上的位点rs1864183(G>A) (P=0.016, HR:0.42, 95%CI:0.21~0.85)对含铂化疗的总生存时间有着显著影响。研究结果提示自噬通路在含铂化疗敏感性中发挥着重要作用,自噬通路基因多态性可能是预测含铂化疗疗效的潜在分子标志物,这可能为临床上肺癌的个体化医疗提供一定的理论基础。     

XPA A23G polymorphism is associated with the elevated response to platinum-based chemotherapy in advanced non-small cell lung cancer

DNA repair capacity （DRC） is correlated with sensitivity of cancer cells toward platinum-based chemotherapy. We hypothesize that genetic polymorphisms in DNA repair gene XPA （xeroderma pigmentosum group A） and XPG （xeroderma pigmentosum group G） （ERCC5, excision repair cross-complementation group 5）, which result in inter-individual differences in DNA repair efficiency, may predict clinical response to platinum agents in advanced non-small cell lung cancer （NSCLC） patients. In this study, we find that the A → G change of XPA A23G polymorphism significantly increased response to platinum-based chemotherapy. Polymorphism in XPG His46His was associated with a decreased treatment response, but was not statistically significant.     

Elevated exhalation of hydrogen peroxide in patients with non-small cell lung cancer is not affected by chemotherapy

Objectives: Reactive oxygen species, which are implicated in the process of carcinogenesis, are also responsible for cell death during chemotherapy (CHT). Therefore, the aim of the study was to evaluate exhaled H₂O₂ levels in non-small cell lung cancer (NSCLC) patients before and after CHT.

Methods: Thirty patients (age 61.3?±?9.3 years) with advanced NSCLC (stage IIIB–IV) and 15 age-matched healthy cigarette smokers were enrolled into the study. Patients received four cycles of cisplatin or carboplatin with vinorelbine every three weeks. Before and after the first, second, and fourth cycle, the concentration of H₂O₂ in exhaled breath condensate was measured with respect to treatment response.

Results: At the baseline, NSCLC patients exhaled 3.8 times more H₂O₂ than the control group (0.49?±?0.14 vs. 0.13?±?0.03?µmol/L, P?2O₂ levels independent of the treatment response (partial remission vs. progressive disease). Pre- and post-CHT cycles of H₂O₂ levels generally correlated positively.

Discussion: The study demonstrated the occurrence of oxidative stress in the airways of advanced NSCLC patients. Exhaled H₂O₂ level was not affected by CHT and independent of treatment results and changes in the number of circulating neutrophils.     

Drug-induced cell cycle perturbation in chemotherapy of patients with non-small cell lung cancer

To observe in vivo cell cycle perturbation in the chemotherapy of lung cancer, tumour cell kinetics during the first course of chemotherapy were measured in seven patients with histologically-verified non-small cell lung cancer. The tumour cells were aspirated from six lymph nodes and one subcutaneous nodule both prior to treatment and twice weekly after the administration of chemotherapeutic agents. The nuclear DNA content of aspirated tumour cells was measured with a scanning microdensitometer at a wavelength of 550 nm after the modified Feulgen reaction. The cell population in cell cycle was estimated with a cumulated percentage scale. Marked cell cycle perturbation occurred within one week after initiation of chemotherapy. There was a decrease in the G1 cell population, from 70.6 +/- 9.1% to 26.1 +/- 11.4%, and a corresponding increase of cells in G2-M phase, from 21.4 +/- 8.7% to 63.7 +/- 10.0%. The proportion of cells in S phase was slightly increased from 8.0 +/- 1.5% to 10.1 +/- 3.2% during this period. The degree of cell cycle changes was unrelated to the clinical response to chemotherapy.     

MiRNA调控非小细胞肺癌转移的研究进展

微小RNA是内源性的非编码小RNA分子,通过与靶mRNA的结合在转录后水平调控基因的表达,从而参与众多生命活动的调控。NSCLC是严重威胁人类健康的恶性肿瘤,侵袭转移是其主要特征,也是其治疗失败和死亡的主要原因。miRNA可以通过促进上皮-间质转化、金属基质蛋白酶表达,以及血管生成来促进NSCLC转移,而且miRNA在调节肺癌干细胞特性中也发挥着重要作用。转移相关的miRNA已成为肺癌靶向治疗的新靶点。     

Finding biomarkers for non-small cell lung cancer diagnosis and prognosis

Despite of several decades of efforts,lung cancer remains one of most deadly diseases,with a 5-year survival rate approximately 15％ worldwide.In China,the situation is even worse.Although there is no o...     

非小细胞肺癌靶向治疗的临床前研究进展

针对表皮生长因子受体（EGFR）和血管生成（angiogenesis）信号通路的靶向治疗已经在晚期非小细胞肺癌的治疗上取得成功,但由于抗药性的存在,大多数晚期患者的生存时间仍然提高有限。继发性的EGFR T790M突变和原癌基因肝细胞生长因子受体（MET）的扩增被鉴定为两种主要的抗药机制。最近转化生长因子-β（TGF-β）/白介素-6信号通路被报道能介导选择性和适应性地对erlotinib的抗药。另一方面,Kras突变所致肺癌的靶向治疗方面也取得了一些进展。双重抑制磷脂酰肌醇3-激酶（PI3K）和促分裂素原活化蛋白激酶激酶（MEK）信号通路可导致Kras突变肿瘤的显著消退,联合抑制SRC、PI3K和MEK可使丝氨酸/苏氨酸蛋白激酶11（Lkb1）缺失,Kras突变的肺癌小鼠的肿瘤明显消退,抑制核因子-κB（NF-κB）信号通路导致p53缺失,Kras突变的肿瘤发展显著减慢。这些发现都为发展非小细胞肺癌患者的靶向治疗提供了有力的支持。     

Peroxiredoxin-I is an autoimmunogenic tumor antigen in non-small cell lung cancer

In eukaryotic cells, peroxiredoxins are both antioxidants and regulators of H(2)O(2)-mediated signaling. We previously found that peroxiredoxin-I (Prx-I) was overexpressed in non-small cell lung cancer (NSCLC) tissue. Since overexpressed protein can induce a humoral immune response, we examined whether serum from NSCLC patients exhibited immunoreactivity against Prx-I using Western blotting. We found that 25 (47%) of 53 NSCLC patients tested had autoantibodies against Prx-I in their sera, whereas such activity was detected in 4 (8%) sera from 50 healthy subjects. Prx-I itself was detected in the sera from 18 (34%) of 53 NSCLC patients but in only 1 (2%) serum from 50 controls. Moreover, 17% of NSCLC sera were positive to both Prx-I antibody and antigen but none in control sera. The data indicate both Prx-I autoantibody and circulating antigen are potential biomarkers for use in serological diagnosis of NSCLC. Interestingly enough, we found that Prx-I was secreted by lung adenocarcinoma cells (A549) but not by non-cancer lung cells (BEAS 2B) or breast cancer cells (MCF7). This cell culture study suggests the possibility of Prx-I secretion from NSCLC tumor tissues.     

Serum interleukin-17 as a diagnostic and prognostic marker for non-small cell lung cancer

Abstract

Objective: The aim of this study was to explore the clinical role of serum interleukin (IL)-17 in patients with non-small-cell lung cancer (NSCLC).

Materials and method: Serum specimens from 128 patients with NSCLC and 60 healthy controls were collected. The concentrations of IL-17 were measured using enzyme-linked immunosorbent assay.

Results: Serum IL-17 levels were higher in the NSCLC group in comparison with the control group (p?<?0.01). With a cut-off value of 16?pg/ml, IL-17 showed a good diagnostic performance for NSCLC. Multivariate survival analysis indicated that IL-17 was an independent prognostic factor in NSCLC.

Conclusion: Measurement of IL-17 might be a useful diagnostic and prognostic value for patients with NSCLC.     

Target volume definition for staple line recurrences of non-small cell lung cancer

BackgroundStaple line (SL) recurrences of non-small cell lung cancer (NSCLC) are commonly treated with radiotherapy (RT), but the target volume definition — whole SL versus focused on recurrence — is unclear. The aim of the study was to determine the appropriate target volume for RT of SL recurrences.Materials and methodsTwenty-two consecutive patients (20 stage I, 2 stage II) treated with salvage RT for SL recurrences were retrospectively analyzed. Imaging features at the time of SL recurrence were evaluated to guide target volume definition.ResultsSurgeryAll patients had complete tumor resection (wedge resection in 10 (45%) and lobectomy in 12 (55%) patients). 14 (64%) patients had risk factors for recurrence, including surgical margins ≤ 2 cm, angiolymphatic and visceral pleural invasion.Salvage RTAfter a median 26 months (9–67), all 22 patients developed SL recurrence which was metabolically active on PET in all and biopsy-confirmed in 18/22 (82%) patients. All patients underwent RT targeting the location of the SL recurrence only. 13/22 (59%) patients had additional PE T-negative nodular or linear SL changes that were not included in the irradiated volume.Recurrence after RTAfter a median 17 months (9–34) 10/22 (45%) patients recurred either regionally 6/10 (60%), in the lungs 4/10 (40%) or distally 3/10 (30%). No patient recurred at the SL. Two-year overall and disease-free survival rates after RT were 71% and 65%, respectively.ConclusionRT to SL recurrences alone results in excellent local control. Additional treatment to reduce regional and distant recurrences should be considered.     

MiR-491在非小细胞肺癌发生发展过程中的作用机制研究

人类miR-491 (has-miR-491)是一段长度为84 bp的微小RNA,可以通过与靶mRNA的3'端非翻译区(3'-untranslated region,3'-UTR)结合,降解靶mRNA或抑制其翻译,在转录后水平对肿瘤细胞基因表达进行调控,影响肿瘤发展进程.非小细胞肺癌(non-small lang ca...     

厚朴酚联合吉非替尼对A549非小细胞肺癌细胞的干预作用*

目的: 探讨厚朴酚与吉非替尼协同影响非小细胞肺癌A549细胞的作用。方法: 以浓度为6.25～500 μmol/L厚朴酚、0.625～100 μmol/L吉非替尼分别处理A549细胞24 h,CCK-8实验检测细胞活力 (n=3),选24 h及100 μmol/L厚朴酚与5 μmol/L吉非替尼作后续处理(n=3);采用对照组、厚朴酚组、吉非替尼组和厚朴酚+吉非替尼组的析因分析设计;克隆形成检测细胞增殖;蛋白印迹测蛋白表达;流式细胞术检测细胞凋亡及分选CD44⁺和CD133⁺细胞。结果: 与对照组比,厚朴酚和吉非替尼组的克隆形成率显著降低(P＜0.05);凋亡率显著升高(P＜ 0.05);CD44⁺和CD133⁺细胞数量显著减少(P＜0.05);Ki67和PCNA及干细胞标记蛋白SOX2和OCT4表达显著下调(P＜0.05);Bax/Bcl-2表达比例显著上调(P＜0.05)。与厚朴酚组或吉非替尼组比较,厚朴酚+吉非替尼组进一步促进了上述改变(P＜0.05),且凋亡率、Bax/Bcl-2、SOX2和OCT4等指标都存在厚朴酚和吉非替尼的交互作用(P＜ 0.05)。结论: 厚朴酚与吉非替尼促进A549细胞凋亡和抑制其干细胞样特性,且联合用药效果优于单一给药。二者对A549细胞的抑癌作用有交互影响。     

Five microRNAs in plasma as novel biomarkers for screening of early-stage non-small cell lung cancer

Background

In order to find novel noninvasive biomarkers with high accuracy for the screening of early-stage non-small cell lung cancer (NSCLC), we investigate the predictive power of 5 microRNAs (miR-20a, miR-145, miR-21, miR223 and miR-221) as potential biomarkers in early-stage NSCLC.

Methods

In training set, 25 early-stage NSCLC patients and 25 matched healthy controls are included to assess the miRNA expression profile between early-stage NSCLC patients and healthy controls by real-time RT-PCR. We found that five of these miRNAs (miR-20a, miR-223, miR-21, miR-221 and miR-145) levels in NSCLC patients were significantly dysregulated compared with the healthy groups and thus were selected to validation set. Therefore, a validation experiment was further performed to investigate the potential predictive power of these five miRNAs based on 126 early-stage NSCLC patients, 42 NCPD patients and 60 healthy controls. The receiver operating characteristic (ROC) curves were generated for the five miRNAs.

Results

ROC curve analyses suggested that these five plasma miRNAs could be promising biomarkers for NSCLC, with relatively high AUC values as follows: miR-20a, 0.89 with 95% CI of [0.85-0.93]; miR-223, 0.94 with 95% CI of [0.91-0.96]; miR-21, 0.77 with 95% CI of [0.71-0.83]; miR-155, 0.92 with 95% CI of [0.89-0.96]; miR-145, 0.77 with 95% CI of [0.71-0.83]. Stratified analyses indicated that plasma miR-20a, miR-223, miR-21 and miR-145 showed better predictive value in smokers than in non-smokers, while miR-155 might be more suitable for non-smokers. In addition, all of these five miRNAs could differentiate NSCLC from controls with a higher accuracy in advanced stage and squamous carcinoma subgroups.

Conclusions

In conclusion, our study suggested that five plasma miRNAs (miR-20a, miR-145, miR-21, miR-223 and miR-221) can be used as promising biomarkers in early screening of NSCLC. Nevertheless, further validation and optimizing improvement should be performed on larger sample to confirm our results.     

非小细胞肺癌患者肠道微生物特征分析

[目的]分析非小细胞肺癌患者与肺部良性病变患者肠道微生物的构成差异,探究特异肠道菌群对非小细胞肺癌发生发展的影响.[方法]收集63例患者粪便样本,非小细胞肺癌患者39例,其中肺腺癌(ADC) 32例,肺鳞癌(SCC)7例,肺部良性病变患者24例,进行16S rDNA测序.[结果]毛螺菌属(Lachnospira)、瘤胃...     

Serum superoxide dismutase,a potential predictor for radiation pneumonitis following chemoradiotherapy in non-small cell lung cancer patients

Purpose: To explore serum superoxide dismutase (SOD) for predicting radiation pneumonitis (RP) in non-small cell lung cancer patients following chemoradiotherapy.

Methods: Serum levels for SOD were measured by enzyme-linked immunosorbent assays prior to radiation therapy (Pre-RT) and post 40 Gy/4 weeks during the treatment (Pos-RT).

Results: SOD concentrations after delivery of 40 Gy/4 weeks was associated with the development of RP. The best predictive ability of SOD was observed for a cut-off value of 56 unit/ml, with a sensitivity of 0.80 (95% CI 0.28–0.99), and a specificity of 0.67 (95% CI 0.43–0.65) (p = 0.040).

Conclusion: Serum SOD may be a potential predictor for RP, which need to be further verified.     

Ring finger protein 19A is overexpressed in non-small cell lung cancer and mediates p53 ubiquitin-degradation to promote cancer growth

The expression pattern, biological functions and the related mechanisms of the ring finger protein 19A (RNF19A) in non-small cell lung cancer (NSCLC) remain poorly understood. This study aimed to explore the role of RNF19A, as well as the underlying potential mechanism, in the development of NSCLC. Here, we found that RNF19A was overexpressed in NSCLC tissues, and RNF19A expression in NSCLC tissue samples was associated with NSCLC carcinogenesis and poor outcome. RNF19A promoted the proliferation of NSCLC cells and inhibited apoptosis. RNF19A reduced p53, p21 and BAX expression and induced Cyclin D1, CDK4, CDK6 and BCL2 expression. The inhibitory effect of RNF19A knockdown on proliferation was partially rescued by p53 silencing. RNF19A interacted with p53, shortened p53 half-life and mediated p53 ubiquitin-degradation. Collectively, we suggest that RNF19A plays a critical oncogenic role in lung carcinogenesis by disrupting the function of p53. RNF19A may serve as a new biomarker and/or target for NSCLC management.     

Orally active microtubule-targeting agent,MPT0B271, for the treatment of human non-small cell lung cancer,alone and in combination with erlotinib

Microtubule-binding agents, such as taxanes and vinca alkaloids, are used in the treatment of cancer. The limitations of these treatments, such as resistance to therapy and the need for intravenous administration, have encouraged the development of new agents. MPT0B271 (N-[1-(4-Methoxy-benzenesulfonyl)-2,3-dihydro-1H-indol-7-yl]-1-oxy-isonicotinamide), an orally active microtubule-targeting agent, is a completely synthetic compound that possesses potent anticancer effects in vitro and in vivo. Tubulin polymerization assay and immunofluorescence experiment showed that MPT0B271 caused depolymerization of tubulin at both molecular and cellular levels. MPT0B271 reduced cell growth and viability at nanomolar concentrations in numerous cancer cell lines, including a multidrug-resistant cancer cell line NCI/ADR-RES. Further studies indicated that MPT0B271 is not a substrate of P-glycoprotein (P-gp), as determined by flow cytometric analysis of rhodamine-123 (Rh-123) dye efflux and the calcein acetoxymethyl ester (calcein AM) assay. MPT0B271 also caused G2/M cell-cycle arrest, accompanied by the up-regulation of cyclin B1, p-Thr161 Cdc2/p34, serine/threonine kinases polo-like kinase 1, aurora kinase A and B and the downregulation of Cdc25C and p-Tyr15 Cdc2/p34 protein levels. The appearance of MPM2 and the nuclear translocation of cyclin B1 denoted M phase arrest in MPT0B271-treated cells. Moreover, MPT0B271 induced cell apoptosis in a concentration-dependent manner; it also reduced the expression of Bcl-2, Bcl-xL, and Mcl-1 and increased the cleavage of caspase-3 and -7 and poly (ADP-ribose) polymerase (PARP). Finally, this study demonstrated that MPT0B271 in combination with erlotinib significantly inhibits the growth of the human non-small cell lung cancer A549 cells as compared with erlotinib treatment alone, both in vitro and in vivo. These findings identify MPT0B271 as a promising new tubulin-binding compound for the treatment of various cancers.     

LINC00184 plays an oncogenic role in non-small cell lung cancer via regulation of the miR-524-5p/HMGB2 axis

Non-small cell lung cancer (NSCLC) is the most common type of lung cancer. We aimed to investigate the role of LINC00184 in NSCLC. Migration, proliferation and invasion of NSCLC cells were analysed using the wound healing assay, cell counting kit-8 assay and transwell assay, respectively. Apoptosis and cell cycle were assessed using flow cytometry. Online bioinformatics tools were utilized to predict downstream microRNAs (miRNA) or genes related to LINC00184 expression. The RNA pull-down experiment and luciferase reporter assay were performed to verify the predictions thereof. LINC00184, miR-524-5p, and high mobility group 2 protein (HMGB2) expression levels in NSCLC tissues and cell lines were detected using quantitative real-time polymerase chain reaction. An NSCLC mouse model was constructed for in vivo experiments. LINC00184 overexpression was observed in NSCLC tissues and cell lines and was found to be correlated with poor prognosis. LINC00184 knockdown inhibited cell proliferation, migration and invasion, induced cell cycle arrest and accelerated apoptosis in NSCLC cell lines. LINC00184 suppressed tumour growth and proliferation in NSCLC mouse models and directly targeted the miR-524-5p/HMGB2 axis. Moreover, the expression levels of LINC00184 and HMGB2 were negatively correlated with miR-524-5p expression, whereas LINC00184 expression was positively correlated with HMGB2 expression. LINC00184 affected the cell cycle, proliferation, apoptosis, migration and invasion in NSCLC via regulation of the miR-524-5p/HMGB2 axis.     

小剂量环磷酰胺联合参一胶囊治疗进展期非小细胞肺癌

目的：应用小剂量环磷酰胺（CTX）联合参一胶囊治疗Ⅲ、Ⅳ期非小细胞肺癌,观察其临床疗效及毒副作用。方法：Ⅲ、Ⅳ期非小细胞肺癌患者经标准化、放疗后随机分为两组,治疗组（38例）：CFX 50mg每日一次、参一胶囊20mg日两次口服,三至六个月;对照组（34例）：化疗后不用任何药物治疗。结果：联合治疗组患者外周血血管内皮生长因子（VEGF）表达下降。T淋巴细胞亚群CD4＋及CD4＋／CD8＋升高,病人生存质量及生存期提高。结论：Ⅲ、Ⅳ期NSCLC患者化疗后联合应用小剂量CTX联合参一胶囊治疗是一种安全有效的治疗方法,可以提高患者的生存质量及生存期。