Single Nucleotide Polymorphisms Of Interleukin-8 And Integrin β₃: Study Of Their Effect Upon The Occurrence And Early Lymph Metastasis Of Cervical Cancer

Purpose: Investigating possible interaction between single nucleotide polymorphisms of IL-8-251 A/T and integrinβ3 Leu33Pro (176T/C) and their effect on the occurrence of cervical cancer and lymph metastasis on the early stage in Shanxi .Methods: A retrospective case-control study design was applied for the present study. The case group included 200 paraffin embedded the early stage of cervical cancer tissue samples (48 cases with lymph metastasis and 152 cases without metastasis) and 217 peripheral blood samples (39 cases with lymph metastasis and 178 cases without metastasis) from patients with the early stage of cervical cancer. The control group consisted of 185 normal paraffin embedded cervical tissue samples and 190 of peripheral blood samples from healthy women. Both groups of samples were collected from Shanxi province. Using genomic DNA as templates, we employed Tetra-primer amplification refractory mutation system polymerase chain reaction (Tetra-primer ARMS-PCR) to analyze the genotype distribution and allele frequency of IL-8-251 A/T and integrinβ3Leu33Pro(176T/C). Chi-square test ( 2 test) was used for statistical comparison of genotype distribution and allele frequency between the case group and the control group as well as between the group with metastasis and the group without metastasis. Odds ratio (OR) and 95% confidence interval (CI) indicated relative risk.The effect modified model was used to evaluate the gene-gene interaction of single nucleotide polymorphisms of IL-8-251 A/T and integrinβ3Leu33Pro(176T/C).We also used Enzyme-Linked ImmunoSorbent Assay (ELISA) to determine the IL-8 levels in the serum and evaluated its possible association with single nucleotide polymorphism of IL-8-251 A/T.Results: 1.IL-8 -251A/T single nucleotide polymorphisms include three genotypes: AA, AT and TT.①For IL-8 -251 AA, AT, TT distribution and A, T allele frequency in tissue samples, there is statistical significance (P<0.05) between the case group and the control group as well as between the group with lymph metastasis and the group without metastasis. The risk of genotype AT, TT and T allele for cervical cancer is 2.081, 2.259 and 1.476, respectively. The risk of genotype TT and T allele for lymph metastasis is 4.309 and 2.435, respectively.②For IL-8-251 genotype distribution and allele frequency in peripheral blood samples, there is also statistical significance (P<0.05) between the case group and the control group as well as between the group with lymph metastasis and the group without metastasis. Genotype AT and TT could be the risk factors for cervical cancer (OR values are 2.290 and 2.619, respectively). Compared to A allele, T allele increase the risk of cervical cancer for 1.6 folds,and genotype TT increase the risk of lymph node metastasis for 3 folds. 2.For integrinβ3 Leu33Pro(176T/C) single nucleotide polymorphisms including genotype distribution and allele frequency, there is no statistical significance (P>0.05) between the case group and the control group; However, there is statistical significance (P<0.05) between the group with metastasis and the group without metastasis. The results also indicate Pro/Pro homozygous could increase the risk of lymph node metastasis for 4 folds(OR=3.989,95%CI=1.492~10.662). 3. There is no interaction between single nucleotide polymorphisms of IL-8-251 A/T and intigrinβ3Leu33Pro(176T/C) in the process of lymph metastasis of cervical cancer on the early satge by using effect modified model. 4. The results from ELISA demonstrate that IL-8 levels in serum are significantly different (P<0.05) between the case group and the control group. Higher serum IL-8 level was detected in the case group with lymph metastasis. According to genotype AA,AT,TT,we devided the case group into three groups,three genotypes have different serum IL-8 level(P<0.05) . People with genotype TT have a higher serum IL-8 level.Conclusions: People with genotype TT and T allele at IL-8 -251A/T site may have a higher risk for the occurrence and early lymph metastasis of cervical cancer. Intigrinβ3 Pro/Pro(C/C) homozygous and Pro(C) allele may also increase the risk of early lymph metastasis of cervical cancer, but intigrinβ3Leu33Pro(176T/C) is not related to the occurrence of cervical cancer in Shanxi. Genotype TT may elevate the serum IL-8 level ,higher serum IL-8 level increases the occurrence of cervical cancer and lymph metastasis on the early stage. Monitoring the serum IL-8 level may contribute to clinical foreseeing of the occurrence and early lymph metastasis of cervical cancer.Taken together, the study points out single nucleotide polymorphisms of IL-8 -251A/T and intigrinβ3 Leu33Pro (176T/C) could be used for screening population with high risk of occurrence and early lymph metastasis of cervical cancer, which will provide clues for prediction, primary prevention of cervical cancer.