M.J. Hong

A genetic variation in microRNA target site of KRT81 gene is associated with survival in early stage non-small cell lung cancer

BACKGROUND MicroRNAs (miRNAs) have a key role in carcinogenesis through negative regulation of their target genes. Therefore, genetic variations in miRNAs or their target sites may affect miRNA-mRNA interactions, thereby result in altered expression of target genes. This study was conducted to investigate the associations between single-nucleotide polymorphisms (SNP) located in the miRNA target sites (poly-miRTSs) and survival of patients with early-stage non-small-cell lung cancer (NSCLC). METHODS Using public SNP database and miRNA target sites prediction program, 354 poly-miRTSs were selected for genotyping. Among these, 154 SNPs applicable to Sequenoms MassARRAY platform were investigated in 357 patients. A replication study was carried out on an independent patient population (n = 479). Renilla luciferase assay and reverse transcription-polymerase chain reaction were conducted to examine functional relevance of potentially functional poly-miRTSs. RESULTS Of the 154 SNPs analyzed in a discovery set, 14 SNPs were significantly associated with survival outcomes. Among these, KRT81 rs3660G>C was found to be associated with survival outcomes in the validation cohort. In the combined analysis, patients with the rs3660 GC + CC genotype had a significantly better overall survival compared with those with GG genotype [adjusted hazard ratio (aHR) for OS, 0.65; 95% confidence interval (CI) 0.50-0.85; P = 0.001]. An increased expression of the reporter gene for the C allele of rs3660 compared with the G allele was observed by luciferase assay. Consistently, the C allele was associated with higher relative expression level of KRT81 in tumor tissues. CONCLUSION The rs3660G>C affects KRT81 expression and thus influences survival in early-stage NSCLC. The analysis of the rs3660G>C polymorphism may be useful to identify patients at high risk of a poor disease outcome.BACKGROUND MicroRNAs (miRNAs) have a key role in carcinogenesis through negative regulation of their target genes. Therefore, genetic variations in miRNAs or their target sites may affect miRNA-mRNA interactions, thereby result in altered expression of target genes. This study was conducted to investigate the associations between single-nucleotide polymorphisms (SNP) located in the miRNA target sites (poly-miRTSs) and survival of patients with early-stage non-small-cell lung cancer (NSCLC). METHODS Using public SNP database and miRNA target sites prediction program, 354 poly-miRTSs were selected for genotyping. Among these, 154 SNPs applicable to Sequenoms MassARRAY platform were investigated in 357 patients. A replication study was carried out on an independent patient population (n = 479). Renilla luciferase assay and reverse transcription-polymerase chain reaction were conducted to examine functional relevance of potentially functional poly-miRTSs. RESULTS Of the 154 SNPs analyzed in a discovery set, 14 SNPs were significantly associated with survival outcomes. Among these, KRT81 rs3660G>C was found to be associated with survival outcomes in the validation cohort. In the combined analysis, patients with the rs3660 GC + CC genotype had a significantly better overall survival compared with those with GG genotype [adjusted hazard ratio (aHR) for OS, 0.65; 95% confidence interval (CI) 0.50-0.85; P = 0.001]. An increased expression of the reporter gene for the C allele of rs3660 compared with the G allele was observed by luciferase assay. Consistently, the C allele was associated with higher relative expression level of KRT81 in tumor tissues. CONCLUSION The rs3660G>C affects KRT81 expression and thus influences survival in early-stage NSCLC. The analysis of the rs3660G>C polymorphism may be useful to identify patients at high risk of a poor disease outcome.

155P: PD-L1 polymorphism can predict clinical outcomes of non-small cell lung cancer patients treated with first-line paclitaxel–cisplatin chemotherapy

QuantStudio® 3D Digital PCR System (ABI, USA). For realtime PCR was used a thermal cycler Rotor-Gene 6000 (Qiagen, Germany), PCR was performed in a final volume of 20ml, reaction mixture contained 70mM Tris-HCl (pH 8.8), 16.6mM ammonium sulphate, 0.01% Tween-20, 2mM magnesium chloride, 200 nM of each dNTP, 500 nM of primers, 250 nM of fluorescent probe, 1000 nM of blocking probe, 1.5 units Taq-DNA polymerase. Probes used in fluorescent dyes – FAM and VIC, quenchers – BHQ-1 and BHQ-2. Sets of oligonucleotides were designed and produced by “Testgen Ltd” (Russia). Results: By comparing of the EGFR mutations status in plasma and tumor samples concordance was 88.7% [95% CI 85%, 92%], sensitivity – 83.3% [95% CI 76%, 90%], specificity – 100%, PPV – 100%. The analytical sensitivity of dPCR was 0.1% Analytical specificity – 99.5%. Analytical sensitivity for real-time PCR was 0.2%, the analytical specificity – 99.7%. Conclusions: Digital PCR has demonstrated the best analytical sensitivity, wherein concordance with real-time PCR was 100%. Legal entity responsible for the study: Kazan Clinical Oncology Center, Kazan, RU Funding: Kazan Clinical Oncology Center, Kazan, RU Disclosure: All authors have declared no conflicts of interest.

108P: Genetic polymorphisms in glycolytic pathway are associated with the prognosis of patients with early stage non-small cell lung cancer

J.E. Choi1, S.K. Do2, M.J. Hong1, J.H. Lee2, J.Y. Park3. 1 Cell and Matrix Research Institute, Kyungpook National University School of Medicine, Daegu, Republic of Korea, 2 Departments of Biochemistry and Cell Biology, Kyungpook National University School of Medicine, Daegu, Republic of Korea, 3 Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu, Republic of Korea

107P: A functional polymorphism located in intron of EGFR affects survival outcome of early-stage non-small cell lung cancer

Background: Radiation pneumonitis (RP) is the most frequent acute pulmonary toxicity following stereotactic body radiation therapy (SBRT) for lung cancer. Here we investigate clinical and dosimetric factors associated with symptomatic RP in stage I non-small cell lung cancer (NSCLC) patients treated with SBRT. Methods: A total of 67 patients with stage I NSCLC who received SBRT at our institution were enrolled and their clinicopathological parameters and dosimetric parameters were recorded and analyzed. Results: The median follow-up period was 26.4 months (range, 7–48 months). In univariable analysis, tumor size (P = 0.041), mean lung dose (MLD; P = 0.028), V2.5 (P = 0.024), V5 (P = 0.014), V10 (P = 0.004), V20 (P = 0.024), V30 (P = 0.020), V40 (P = 0.040), V50 (P = 0.040) were associated with symptomatic RP. In multivariable logistic regression analysis, V10 (P = 0.049) was significantly associated with symptomatic RP. Conclusions: In conclusion, this study found that tumor size, MLD and V2.5–50 were risk factors markedly associated with symptomatic RP.V10 was the most significant factor and planning constraints should be optimized to minimize the lung dose V10.For both central and peripheral stage I lung cancer, rates of RP grade 2 was low after SBRT with appropriate fraction dose and it has no correlation with the location of radiation field. Clinical trial identification: ChiCTR-OPN-15006864 (Chinese Clinical Trial Registry) Legal entity responsible for the study: N/A Funding: N/A Disclosure: All authors have declared no conflicts of interest.

185PASSOCIATION OF POLYMORPHISMS IN THE MICRORNA TARGET SITES AND SURVIVAL OF PATIENTS IN EARLY-STAGE NON-SMALL-CELL LUNG CANCER

ABSTRACT Aim: MicroRNAs (miRNAs) have a key role in carcinogenesis through the negatve regulation of their target genes. Therefore, genetic variations in miRNAs or miRNA target sites may affect miRNA-mRNA interactions, thereby result in altered expression of target genes. This study was conducted to investigate the associations between single nucleotide polymorphisms (SNPs) located in the miRNA target sites (poly-miRTSs) and survival of patients with early-stage non-small cell lung cancer (NSCLC). Methods: Using public SNP database and miRNA target sites prediction program, 354 poly-miRTSs were selected for genotyping. Among these, only 154 SNPs could be applied to Sequenoms MassARRAY platform and investigated in 357 patients. A replication study was performed on an independent patient population (n = 479). Renilla luciferase assay and reverse transcription-PCR were conducted to examine functional relevance of potentially functional poly-miRTSs. Results: Of the 154 SNPs analyzed in the discovery set, 14 SNPs were significantly associated with survival outcomes. Among these, KRT81 rs3660C > G was found to be associated with survival outcomes in the validation cohort. In combined analysis, patients with the rs3660 GC + CC genotype had a significantly better overall survival (OS) compared with those with GG genotype (adjusted hazard ratio for OS, 0.65; 95% confidence interval 0.50-0.85; p = 0.001). An increased expression of the reporter gene for the C allele of rs3660 compared with the G allele was observed by luciferase assay. Relative expression level of KRT81 in tumor tissues was significantly higher in CC genotype compared with GG or GC genotypes. Conclusions: The rs3660G > C affects KRT81 expression and thus influences survival in early-stage NSCLC. The analysis of the rs3660G > C polymorphism may be useful to identify patients at high risk of a poor disease outcome. Disclosure: All authors have declared no conflicts of interest.