HUBUNGAN LESI PRA KANKER SERVIKS HIGH RISK-HUMAN PAPILLOMAVIRUS POSITIF DENGAN KANKER SERVIKS TERKAIT EKSPRESI mRNA TOLL-LIKE RECEPTOR 7 DAN mRNA TOLL-LIKE RECEPTOR 9

SRI HARTINI, DR., SP.PK., MARS., Prof.dr.Marsetyawan. HNE Soesatyo., M.Sc.,Ph.D.

2015 | Disertasi | S3 Ilmu Kedokteran

Latar belakang : Respon imun memegang peranan penting dalam perjalanan infeksi HR-HPV menjadi kanker serviks terutama respon imun bawaan. Respon imun bawaan dan sekresi sitokin sangat dipengaruhi oleh Toll-like Receptor (TLR). TLR 7 dan TLR 9 merupakan sensor kunci sistem imunitas bawaan melalui pengenalan komponen virus. Tujuan : Mengkaji hubungan mRNA TLR 7 dan mRNA TLR 9 lesi pra kanker serviks HR-HPV positif dengan kanker serviks dan mengetahui distribusi HR-HPV pada ke 2 lesi kanker tersebut. Metode : Penelitian cross sectional terhadap jaringan biopsi kanker serviks dan pra kanker Neoplasia Intraepitelial Serviks (NIS) 2 dan 3. Didapatkan 55 sampel dalam periode Juli 2013 sampai Agustus 2014. Ekspresi mRNA TLR 7, 9 dan GAPDH diperiksa dengan RT-PCR kuantitatif, primer didesain tersendiri serta HPV DNA genotyping diperiksa dengan metode hibridisasi Linnear Array. Kuantifikasi relatif ekspresi mRNA TLR dinyatakan sebagai ratio terhadap GAPDH. Normalitas data TLR diuji dengan uji Saphiro-Wilk. Hubungan mRNA TLR 7 dan mRNA TLR 9 dengan pra kanker dan kanker serviks serta positivitas HR-HPV dilakukan dengan uji independent t test untuk beda rerata 2 kategori, batas kemaknaan p <0.05, CI 95%. Beda rerata lebih dari 2 kategori dilakukan dengan uji Anova dan multiple comparisons. Hubungan HR-HPV dengan diagnosis diuji dengan uji Chi-Square. Hasil : Didapatkan diagnosis Squamous Ca (58,2%), NIS 2/3 (23,6%) dan Adeno Ca (18,2%). Rerata ekspresi mRNA TLR7 kanker dan pra kanker tidak berbeda bermakna (p= 0,109). Ekspresi TLR7 NIS 2/3 lebih tinggi bermakna dibanding Squamous Ca dan Adeno Ca (p=0,011). Ekspresi mRNA TLR7 HPV (+) tidak berbeda bermakna dengan HPV (-). Ekspresi mRNA TLR 7 tidak berkaitan baik dengan HPV 16 maupun HPV 18 namun ekspresinya pada HPV 18 lebih rendah bermakna dari HPV multi tipe (p=0,022). mRNA TLR9 pada Adeno Ca lebih tinggi bermakna dibandingkan pra kanker (p=0,044). Ekspresi mRNA TLR9 lebih rendah pada kelomok HPV (+) dibanding HPV (-) dengan p= 0,011 dan ekspresinya lebih rendah pada kelomok HPV16 (+) dibanding HPV 16 (-) dengan p=0,002. Ekspresi mRNA TLR 9 pada HPV 18 tidak berbeda bermakna dengan HPV multi tipe. Didapatkan distribusi frekuensi tipe HPV 18,16,45,52 dan HPV multi tipe. Positivitas HPV lebih tinggi pada kanker dibanding pra kanker (p=0,05). HPV 16 (+) lebih sering pada Squamous Ca, HPV 18 (+) lebih sering pada Adeno Ca. Kesimpulan : TLR7 menyiratkan lebih berperan sebagai pro inflamasi, sementara hasil ini mendukung dugaan TLR9 lebih berperan sebagai pro tumor atau meningkatkan pertumbuhan tumor. HPV 16 menekan transkripsi TLR 9 sebagai upaya menghindar dari sistem imun. Dominasi tipe HPV adalah tipe 16 dan 18. Selain itu didapatkan tipe HR- HPV 45 dan 52 serta HPV multi tipe yang cukup tinggi (27,3%)

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%)

Kata Kunci : TLR 7, TLR 9, HR-HPV, Kanker serviks