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

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

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

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

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

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

Differential Impact of IL-10 Expression on Survival and Relapse between HPV16-Positive and -Negative Oral Squamous Cell Carcinomas

<div><p>Human papillomavirus (HPV) is a risk factor in a subset of oropharyngeal cancer; however, the contribution of HPV in the malignancy of oral squamous cell carcinomas (OSCC) is not fully understood in Taiwanese. Herein, 61 patients with no risk factors and 117 patients with one or more risk factors were enrolled in this study. HPV16/18 infection rate in non-smokers, non-drinkers and non-betel quid chewers was higher than their counterparts. The development of HPV-infected cancer has been shown to be associated with interleukin-10 (IL-10) expression. To this end, IL-10 mRNA expression in OSCC tumors was evaluated by real-time RT-PCR. Data showed that HPV-positive patients had higher IL-10 mRNA levels than in HPV-negative patients. Kaplan-Meier and Cox-regression analysis indicated that the prognostic significance of IL-10 mRNA on overall survival and relapse free survival was only observed in HPV-positive OSCC, but not in HPV-negative OSCC. Mechanistically, the elevation of IL-10 by E6 was responsible for increased colony formation and migration capability in OSCC cells. Therefore, we suggest that IL-10 induced by E6 promotes cell growth and migration capability and consequent poor survival and relapse in HPV-positive OSCC.</p> </div

Relationships between HPV16, HPV18 and HPV16/18 infection and clinical parameters in OSCC patients.

1<p>Other tumor sites includes plate (n = 1), gingival (n = 8), and lip (n = 9).</p>*<p>p<0.05;</p>#<p>p = 0.05.</p