CERVICAL CANCER LESIONS RELATED HIGH RISK PRE-Human Papillomavirus Cervical Cancer ASSOCIATED WITH POSITIVE mRNA Expression of toll-like receptor 7 AND mRNA toll-like receptor 9
ABSTRACT: Background: The immune response, especially the innate immune response plays an important role in the course of HR-HPV infection into cervical cancer. The innate immune response and cytokine secretion was strongly influenced by Toll-like Receptor (TLR). TLR7 and TLR9 is a key role of innate immune system through the introduction of viral components. Objective: To assess the relationship of mRNA TLR7 and mRNA TLR9 in precancerous cervical lesions HR-HPV (+) and cervical cancer subjects and determine the distribution of HR-HPV in those lesions. Methods: This study is a cross sectional study on cervical cancer biopsy tissue and cervical intraepithelial neoplasia (CIN) 2 and 3. Fifty five (55) samples were obtained from July 2013 to August 2014. The mRNA expression of TLR7, 9 and GAPDH were analyzed by quantitative Real Time PCR, primer was designed separately and genotyping of HPV DNA was analysed by Linnear Array hybridization. Relative quantification of mRNA TLRs were expressed as a ratio of its expression to GAPDH. Normality of TLR data was tested with Shapiro-Wilk test. Correlation between mRNA TLR7 and TLR9 in pre-cancer and cervical cancer as well as positivity of HR-HPV test performed by independent t test, the limit of significance p <0.05, confidence interval / CI 95%. Mean difference of more than 2 categories performed with ANOVA and multiple comparisons test. Correlation between HR-HPV and the diagnosis was analysed by Chi-Square. Results: Distribution of Squamous Ca is 58.2%, CIN 2/3 is 23.6% and Adeno Ca is 18.2%. Mean of mRNA TLR7 expression between cancer and pre-cancerous did not differ significantly with p = 0.109. TLR7 expression in CIN 2/3 was significantly higher than Adeno Ca and Squamous Ca with p = 0.011. expression of mRNA TLR 7 in HPV (+) did not differ significantly with HPV (-). Expression of mRNA TLR7 was not associated either with HPV 16 and HPV 18, but its expression in significantly lower in HPV 18 than the multi HPV types (p = 0.022). mRNA TLR9 expression in Adeno Ca was significantly higher than in pre-cancers (p = 0.044). mRNA TLR9 expression was significantly lower in the group of HPV (+) compared to HPV (-) with p = 0.011 and its expression was lower in HPV16 (+) group compared to HPV 16 (-) group with p = 0.002. mRNA TL9 expression in HPV 18 group was not significantly different with multi type HPV group. This study obtained the distribution of HPV types 18, 16, 45, 52 and HPV multi type. HPV positivity was higher in cancer than pre-cancer with p = 0.05. HPV 16 (+) is frequently found in squamous Ca but HPV 18 (+) is frequently found in Adeno Ca. Conclusions : TLR7 is suggested play an important role in pro-inflammatory activity, while TLR9 plays a role in pro tumors activity or promote tumor growth. HPV 16 can suppress TLR9 transcription in an attempt to escape from the immune system. The major type of HPV are type 16 and 18 followed by HR- HPV types 45 and 52 as well as multi-type HPV (27.3%)
