Interleukin-10 Haplotype May Predict Survival and Relapse in Resected Non-Small Cell Lung Cancer

<div><p>IL-10 is associated with tumor malignancy via immune escape. We hypothesized that IL-10 haplotypes categorized by IL-10 promoter polymorphisms at –1082A>G, –819C>T, and –592C>A might influence IL-10 expression and give rise to non-small cell lung cancer (NSCLC) patients with poor outcomes and relapse. We collected adjacent normal tissues from 385 NSCLC patients to determine IL-10 haplotypes by direct sequencing and polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). Of the 385 tumors, 241 were available to evaluate IL-10 mRNA expression levels by real-time RT-PCR. The influence of IL-10 haplotypes on overall survival (OS) and relapse free survival (RFS) were determined by Kaplan-Meier and multivariate Cox regression analysis. The results showed that IL-10 mRNA levels were significantly higher in tumors with the non-ATA haplotype than with the ATA haplotype (P = 0.004). Patients with the non-ATA haplotype had shorter OS and RFS periods than did patients with the ATA haplotype. This may be associated with the observation that the number of tumor-infiltrating lymphocytes was decreased in the tumors with higher levels of IL-10. Consistently, T cells from the peripheral blood of the patients with non-ATA haplotype were more susceptible to apoptosis and less cytotoxic to tumor cells, compared to those from the patients with ATA haplotype. The results suggest that IL-10 can promote tumor malignancy via promoting T cell apoptosis and tumor cell survival, and IL-10 haplotype evaluated by PCR-RFLP or direct sequencing may be used to predict survival and relapse in resected NSCLC, helping clinicians to make appropriate decisions on treatment of the patients.</p> </div

Kaplan-Meier actuarial analysis of RFS and OS.

<p>According to IL-10 haplotype (A), IL-10 mRNA (B), and the combination of IL-10 haplotype and mRNA (C).</p

Unadjusted and adjusted overall incidence rates and age-specific adjusted incidence of multiple myeloma for the period of 1997–2009.

<p>Unadjusted and adjusted overall incidence rates and age-specific adjusted incidence of multiple myeloma for the period of 1997–2009.</p

Correlations between tumor infiltrating lymphocyte counts and IL-10 haplotype in 87 non-small cell lung cancer patients.

1<p>Two sided Chi-square test.</p

IL-10 mRNA expression (mean ± standard error) in tumor tissue by promoter haplotypes.

<p>IL-10 mRNA expression (mean ± standard error) in tumor tissue by promoter haplotypes.</p

Histological examination of TC-1 tumors in lung following i.v. injection of 1×10⁵ TC-1 cells with injection of IgG antibody or IL-10 neutralize antibody

<p>(<b>20μg per 3 days</b>)<b>.</b> Mice were sacrificed in the 14^th days. Lung metastasis was found in mice injected with IgG antibody (A) and was not found in those with IL-10 neutralize antibody (B).</p

Correlations between IL-10 haplotypes and clinical characteristics in 385 non-small cell lung cancer patients.

<p>SQ, squamous cell carcinomas; AD, adenocarcinomas.</p>1<p>Two sided Chi-square test.</p

Multivariate analysis of the influence of IL-10 haplotype determined from IL10 mRNA on overall survival and relapse free survival in non-small cell lung cancer patients.

<p>Adjusted for age, gender, smoking, tumor type, and stage.</p

Representative immunostaining of CD3⁺ TILs in lung tumors.

<p>(A) Representative of CD3 immunostaining of TILs in lung tumors with low TIL density (<25 CD3⁺/HPF) (left: 100x; right: 400x); (B) TILs presented in lung tumors show high TIL density (≥25 CD3⁺/HPF) (left: 100x; right: 400x).</p

Demographic characteristics of cohorts with and without multiple myeloma.

#<p>p value was evaluated by Student’s <i>t</i>-test.</p