Construction of a prognostic model for non-small-cell lung cancer based on ferroptosis-related genes

We wished to construct a prognostic model based on ferroptosis-related genes and to simultaneously evaluate the performance of the prognostic model and analyze differences between high-risk and low-risk groups at all levels. The gene-expression profiles and relevant clinical data of patients with non-small-cell lung cancer (NSCLC) were downloaded from public databases. Differentially expressed genes (DEGs) were obtained by analyzing differences between cancer tissues and paracancerous tissues, and common genes between DEGs and ferroptosis-related genes were identified as candidate ferroptosis-related genes. Next, a risk-score model was constructed using univariate Cox analysis and least absolute shrinkage and selection operator (Lasso) analysis. According to the median risk score, samples were divided into high-risk and low-risk groups, and a series of bioinformatics analyses were conducted to verify the predictive ability of the model. Single-sample gene set enrichment analysis (ssGSEA) was used to investigate differences in immune status between high-risk and low-risk groups, and differences in gene mutations between the two groups were investigated. A risk-score model was constructed based on 21 ferroptosis-related genes. A Kaplan–Meier curve and receiver operating characteristic curve showed that the model had good prediction ability. Univariate and multivariate Cox analyses revealed that ferroptosis-related genes associated with the prognosis may be used as independent prognostic factors for the overall survival time of NSCLC patients. The pathways enriched with DEGs in low-risk and high-risk groups were analyzed, and the enriched pathways were correlated significantly with immunosuppressive status.     

Effects of cryotherapy or chemotherapy on apoptosis in a non-small-cell lung cancer xenografted into SCID mice

Lung cancers are among the most frequent and the most lethal tumours. They are mainly treated by surgery or by chemotherapy, but in the most advanced stages a local cryotherapy can be proposed as a palliative option for bronchial clearance. This therapy, based on the cytotoxic effects of low temperatures, acts by mechanisms which are not yet totally understood. The aim of this work was to investigate in vivo the biological effects of cryotherapy in a model of human non-small-cell lung cancer. We used a xenograft system: cells from the A549 cell line (adenocarcinoma) were injected subcutaneously into SCID mice. Cryotherapy was performed (three cycles, nitrous oxide cryoprobe). Chemotherapy (intravenous injection of Vinorelbine (Navelbine), 4.8 mg/kg) was used as a control treatment. Tumour nodes were excised at variable time points and studied morphologically. The induction of apoptosis was analysed by immunohistochemical staining of cleaved caspase-3 and by TUNEL. Results showed that cryotherapy was an efficient technique to induce cell death either by necrosis or by apoptosis. Necrosis was found near the cryoprobe impact site and was maximal 2 h after treatment (65%); a second peak was observed after 4 days (77%). Around this central necrotic area, apoptotic cells were found. Apoptosis was maximal after 8 h (47%). Chemotherapy induced apoptosis in a fewer number of cells and this effect was not time-dependent. Taken together, these results demonstrate the differential effects of cryotherapy and chemotherapy in vivo, suggesting different modes of action and the potential benefit to combine them.     

MDR1 single nucleotide polymorphism C3435T in Turkish patients with non-small-cell lung cancer

We assessed whether single nucleotide polymorphisms (SNPs) in MDR1 gene C3435T predicted the outcome of platinum-based chemotherapies and survival in our non small cell lung cancer (NSCLC) patients. A total of 79 non-small cell lung cancer patients were enrolled to study. We determined the MDR1 C3435T single nucleotide gene polymorphisms. Median age was 60years: 91.7% male, 8.9% female. We found that CC, CT, TT genotype and T, C allele frequencies in lung cancer patients as 24.1%, 62%, 13.9% and 44.3%, 55.7%, respectively. Patients with CT genotype had a higher response rate (11.4%) than the other genotypes. However, this difference is not statistically significant (p=0.743). Cox regression analysis for overall survival showed that ECOG PS status 0 (HR PS 1 vs. 0, 5.68 p=0.002; HR of PS 2 vs. 0 is 21.579, p=0.001; HR of PS 3 vs. 0 is 35.35, p=0.001), stage ≤II (HR of stage III vs. I+II is 17.77; p=0.016, HR of stage IV vs. I+II is 26.97, p=0.006), and albumin level ≥3g/dl (HR of albumin <3g/dl vs. ≥3g/dl is 2.46, p=0.044) were the most important prognostic factors (also, time to progression was related to these factors). There was no significant association between the genotypes and clinicopathologic parameters; however, good performance status, early stage and ≥3g/dl albumin level were found to be the most important prognostic factors for overall survival and progression-free survival.     

Phase I trial of chimeric (human-mouse) monoclonal antibody L6 in patients with non-small-cell lung,colon, and breast cancer

We report a single institution phase I trial of chimeric (mouse-human) monoclonal antibody (chL6) directed against a tumor-associated cell surface antigen expressed in non-small cell lung, colon, and breast cancer. The results of the study were contrasted with a previous trial of murine L6. ChL6 was administered intravenously to 18 patients with advanced cancer as a single, 4–16 infusion in doses ranging from 350 mg/m² to 700 mg/m². One patient received four weekly doses of 350 mg/m². Patients were followed for side effects, localization of antibody to tumor cells, pharmacokinetics and the development of antibodies against chL6. Side effects associated with treatment were chills, fever, and nausea, which lasted 24–48 hours. Platelet count and absolute leukocyte count fell immediately after treatment, but returned to pretreatment levels by day 7. Localization of chL6 to tumor cells in vivo was seen at 350 mg/m² and saturation at 700 mg/m² and 350 mg/m² per week×4. The pharmacokinetics of this antibody appeared similar to its murine analogue. Human antibodies against chL6 were detected in only 4 of 18 patients. These antibodies were directed against murine variable regent and their titers were lower than those occurring in most patients who received murine L6 in an earlier trial. No tumor reductions were seen. Chimeric L6 appears to be a suitable antibody for delivering anti-tumor agents because of its low immunogenicity and favorable in vivo tumor binding characteristics.     

Effect of polymorphisms in XPD on clinical outcomes of platinum-based chemotherapy for Chinese non-small cell lung cancer patients

Purpose

Xeroderma pigmentosum group D (XPD) codes for a DNA helicase involved in nucleotide excision repair that removes platinum-induced DNA damage. Genetic polymorphisms of XPD may affect DNA repair capacity and lead to individual differences in the outcome of patients after chemotherapy. This study aims to identify whether XPD polymorphisms affect clinical efficacy among advanced non-small cell lung cancer (NSCLC) patients treated with platinum-based chemotherapy.

Experimental Design

353 stage III-IV NSCLC patients receiving platinum-based chemotherapy as the first-line treatment were enrolled in this study. Four potentially functional XPD polymorphisms (Arg¹⁵⁶Arg, Asp³¹²Asn, Asp⁷¹¹Asp and Lys⁷⁵¹Gln) were genotyped by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry or PCR-based sequencing.

Results

Variant genotypes of XPD Asp³¹²Asn, Asp⁷¹¹Asp and Lys⁷⁵¹Gln were significantly associated with poorer NSCLC survival (P = 0.006, 0.006, 0.014, respectively, by log-rank test). The most common haplotype GCA (in order of Asp³¹²Asn, Asp⁷¹¹Asp and Lys⁷⁵¹Gln) also exhibited significant risk effect on NSCLC survival (log-rank P = 0.001). This effect was more predominant for patients with stage IIIB disease (P = 2.21×10⁻⁴, log-rank test). Increased risks for variant haplotypes of XPD were also observed among patients with performance status of 0–1 and patients with adenocarcinoma. However, no significant associations were found between these polymorphisms, chemotherapy response and PFS.

Conclusions

Our study provides evidence for the predictive role of XPD Asp³¹²Asn, Asp⁷¹¹Asp and Lys⁷⁵¹Gln polymorphisms/haplotype on NSCLC prognosis in inoperable advanced NSCLC patients treated with platinum-based chemotherapy.     

Application of proteomics in non-small-cell lung cancer

Non-small-cell lung cancer (NSCLC) is a heterogeneous disease with diverse pathological features. Clinical proteomics allows the discovery of molecular markers and new therapeutic targets for this most prevalent type of lung cancer. Some of them may be used to detect early lung cancer, while others may serve as predictive markers of resistance to different therapies. Therapeutic targets and prognostic markers in NSCLC have also been discovered. These proteomics biomarkers may help to pair the individual NSCLC patient with the best treatment option. Despite the fact that implementation of these biomarkers in the clinic appears to be scarce, the recently launched Precision Medicine Initiative may encourage their translation into clinical practice.     

Effect of Smac and Taxol on non-small-cell lung cancer

A series of structurally unique second mitochondria-derived activator of caspases （Smacs） that act as antagonists of the inhibitor of apoptosis proteins （IAPs） directly have been discovered. They play crucial roles in mitochondrial apoptosis pathways and promote chemotherapy-induced apoptosis. In this study, we constructed a eukaryotic expression vector pcDNA3.1/Smac and transfected it into A549 human lung cancer cells. Then we analyzed the cell invasive and cloning ability, as well as cell apoptosis induced by Taxol. The results showed that over-expressed Smac significantly inhibited A549 cell invasive and cloning ability and promoted apoptosis following Taxol treatment. This finding provides a potential approach for the biological therapy of lung cancer.     

CYFRA21-1 can predict the sensitivity to chemoradiotherapy of non-small-cell lung carcinoma

Background: The increasing panel of systemic therapies enables the individual management of lung cancer patients, even in advanced stages. However, predictive tools indicating the efficacy of chemoradiotherapy (CRT) are badly needed.

Aims: To determine the tumour markers for predicting the therapeutic effect in non-small-cell lung carcinoma (NSCLC) patients treated with CRT.

Methods: The serum levels of cytokeratin 19 fragment antigen 21-1 (CYFRA21-1), neurone-specific enolase (NSE) and carcinoembryonic antigen (CEA) were measured before CRT by enzyme-linked immunosorbent assays, while the tumour responses were assessed according to the World Health Organization (WHO) response criteria. The relationships between pretreatment expression of CYFRA21-1, NSE, CEA and the effectiveness of CRT were analysed.

Results: The complete response (CR) rate of the primary tumours estimated by computed tomography in patients with high levels of CYFRA21-1 was 2.9% (2/68) while in cases with low CYFRA21-1 it was 20.3% (12/59) (p?=?0.005). The effective rates (CR+PR) in CYFRA21-1 high and low groups were 52.9% (36/68) and 72.9% (43/59), respectively (p?=?0.022).

Conclusions: CYFRA21-1 may be a reliable surrogate marker of CRT efficacy in patients with NSCLC.     

RhoC和Ki67在非小细胞肺癌中的表达及意义

目的探讨RhoC、Ki67在非小细胞肺癌中的表达状况及其与临床病理学参数间的关系。方法利用免疫组织化学(ElivisionTMplus法),检测肺癌组织芯片中151例非小细胞肺癌患者RhoC、Ki67的表达情况。结果肺癌组织的RhoC蛋白阳性表达率为59.60%(90/151),相应癌旁组织RhoC阳性表达率为32.7%(16/49),差别有显著意义(P<0.05)。RhoC在TNMIII/IV期的NSCLC组织中阳性表达率为68.9%(51/74),明显高于在Ⅰ/II期NSCLC组织中阳性表达率50.6%(39/77,P<0.05);RhoC在出现淋巴结转移的表达率为49.2%(32/65),显著高于没有淋巴结转移的阳性表达率67.4%(58/86,P<0.05);非小细胞肺癌中RhoC高表达同Ki-67高表达呈显著正相关关系(χ2=21.634,r=0.377,P<0.01)。结论RhoC的高表达与非小细胞肺癌的分期,淋巴结转移及增殖有关。     

小细胞肺癌患者外周血淋巴细胞亚群分析

目的 探究化疗对小细胞肺癌(small cell lung cancer,SCLC)患者免疫功能的影响。 方法 选择2013年1月到2018年12月我院收治的95例小细胞肺癌患者为研究对象。患者第一周期、第二周期化疗前采用流式细胞术检测患者外周血淋巴细胞亚群水平,分别按照不同疗效及不同化疗方案对患者外周血淋巴细胞亚群进行比较。 结果 (1)化疗后,95例患者CD3+、CD4+、CD8+细胞平均值增加,CD19+、γδT细胞平均值减少,差异均有统计学意义(均P+、CD8+细胞平均值增加,CD19+细胞减少,差异有统计学意义(均P0.05)。(3)依托泊苷联合顺铂(EP)方案组化疗后患者CD3+、CD8+细胞平均值增多,CD19+细胞减少,差异均有统计学意义(均P0.05)。 结论 化疗可以调节小细胞肺癌患者的免疫功能,增强细胞免疫,降低体液免疫,其中EC方案对患者细胞免疫的增强作用较为显著。     

WT1 peptide-based immunotherapy for patients with lung cancer: report of two cases

The Wilms' tumor gene WT1 is overexpressed in various types of solid tumors, including lung and breast cancer and WT1 protein is a tumor antigen for these malignancies. In phase I clinical trials of WT1 peptide-based cancer immunotherapy, two patients with advanced lung cancer were intradermally injected with 0.3 mg of an HLA-A*2402-restricted, 9-mer WT1 peptide emulsified with Montanide ISA51 adjuvant. Consecutive WT1 vaccination at 2-week intervals resulted in a reduction in tumor markers such as chorio-embryonic antigen (CEA) and sialyl Lewis (x) (SLX) and by a transient decrease in tumor size. No adverse effects except for local erythema at the injection sites of WT1 vaccine were observed. These results provided us with the first clinical evidence demonstrating that WT1 peptide-based immunotherapy should be a promising treatment for patients with lung cancer.     

An investigation of the relationship between SULT1A1 Arg213His polymorphism and lung cancer susceptibility in a Turkish population

Human sulfotransferase 1A1 (SULT1A1), the most expressed isoform of the phenol SULT1 subfamily, is an important member of sulfotransferase superfamily. A transition, G to A at position 638, in SULT1A1 gene, results in Arg²¹³His change. This single nucleotide polymorphism reduces the activity and thermostability of SULT1A1 enzyme. Thus, in the present study the relationship between SULT1A1 Arg²¹³His polymorphism and lung cancer was investigated. One hundred and six case and 271 control samples were studied using PCR‐RFLP. There was no significant difference in genotype and allele distribution between lung cancer and control populations (p = 0.07; p = 0.06, respectively). Compared with the SULT1A1*1/SULT1A1*1 genotype the variant SULT1A1 genotype (SULT1A1*1/SULT1A1*2 or SULT1A1*2/SULT1A1*2) was associated with a significantly increased lung cancer risk in cases (p = 0.027). In male populations, there was no significant difference between case and controls (p = 0.313). In female populations, however, this difference was found to be significant (p = 0.04). In smoker and non‐smoker populations, no significant relationship was evident between lung cancer and control population (p = 0.170, p = 0.065, respectively). Statistical analyses of histological types of lung cancer in comparison with the control individuals indicated a significant difference between SULT1A1 Arg²¹³His polymorphism and SCC (p = 0.027) and other types of cancer (p = 0.037), except SMCC (p = 0.854). Copyright © 2009 John Wiley & Sons, Ltd.     

Nanoparticle-biomolecular corona: A new approach for the early detection of non-small-cell lung cancer

Lung cancer (LC) is the most common type of cancer and the second cause of death worldwide in men and women after cardiovascular diseases. Non-small-cell lung cancer (NSCLC) is the most frequent type of LC occurring in 85% of cases. Developing new methods for early detection of NSCLC could substantially increase the chances of survival and, therefore, is an urgent task for current research. Nowadays, explosion in nanotechnology offers unprecedented opportunities for therapeutics and diagnosis applications. In this context, exploiting the bio-nano-interactions between nanoparticles (NPs) and biological fluids is an emerging field of research. Upon contact with biofluids, NPs are covered by a biomolecular coating referred to as “biomolecular corona” (BC). In this study, we exploited BC for discriminating between NSCLC patients and healthy volunteers. Blood samples from 10 NSCLC patients and 5 subjects without malignancy were allowed to interact with negatively charged lipid NPs, leading to the formation of a BC at the NP surface. After isolation, BCs were characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). We found that the BCs of NSCLC patients was significantly different from that of healthy individuals. Statistical analysis of SDS-PAGE results allowed discriminating between NSCLC cancer patients and healthy subjects with 80% specificity, 80% sensitivity and a total discriminate correctness rate of 80%. While the results of the present investigation cannot be conclusive due to the small size of the data set, we have shown that exploitation of the BC is a promising approach for the early diagnosis of NSCLC.     

ERCC1多态性与肺癌铂类化疗耐药的关系研究

目的：研究DNA切除修复交叉互补基因1（excision repair cross-complementing gene1,ERCC1）单核苷酸多态性与非小细胞肺癌铂类药物化疗敏感性的关系。方法：应用基因测序方法检测89例以铂类药物为主要化疗方案的非小细胞肺癌患者的ERCC1 Asn118Asn基因型,,比较不同基因型与化疗疗效的关系。结果：89例患者化疗总有效率为29.2%。携带ERCC1 CC基因型、含至少一个变异基因型（TC和TT基因型）患者的有效率分别为38.5%和61.5%（X2=2.151,p=0.142）,基因型在化疗有效组和无效组之间的分布无差异（p〉0.05）。结论：ERCC1Asn118Asn单核苷酸多态性可能与非小细胞肺癌对铂类药物化疗的敏感性无关。     

Epigenetic silencing of human T (brachyury homologue) gene in non-small-cell lung cancer

Early detection of lung cancer is challenging due to a lack of adequate biomarkers. To discover novel tumor suppressor genes (TSGs) silenced by aberrant promoter methylation, we analyzed the gene expression profiles of two lung adenocarcinoma cell lines using pharmacologic-unmasking and subsequent microarray-analysis. Among 617 genes upregulated, we selected 30 genes and investigated the methylation status of their promoters by bisulfite sequencing analysis. Aberrant methylation was detected in four genes (CRABP2, NOEY2, T, MAP2K3) in at least one lung adenocarcinoma cell lines. Furthermore, the T promoter was methylated in 60% of primary lung adenocarcinomas versus 13% of non-malignant lung tissues. Conversely, RT-PCR analysis revealed T expression was low in lung tumors, while high in normal tissues. In addition, no non-synonymous mutations related to gene silencing were found. While further analysis is warranted, our results suggest that T has the potential to be a novel candidate TSG in lung cancer.