The Relationship Between Methylation Of FHIT, RUNX3and Lung Carcinoma

Objective：In this study we collected50NSCLC tissue samples and50samples of homologousnormal lung tissue which were5cm from the lung cancer tissue. we got the methylationstatus of the FHIT and RUNX3gene promoter, using MSP (methylation specific PCR)to amplify the target gene fragment，then extracted by3%agarose gel electrophoresis.According to the genes methylation status among these100samples, we analyze theclinical and pathological relationship between the methylation of FHIT、RUNX3withthe development of non-small cell lung cancer, in order to provide information relevantresearch evidence for early diagnosis of lung cancer.Method:1. We collected50non-small-cell lung cancer tissue samples and50homologousnormal lung tissue samples, the specimens were preserved with low-temperaturefreezer;2. Genomic DNAextraction kit were utilized to extract DNA of the100specimens.3. Using MSP (methylation specific PCR) to amplify the DNA.4. The DNAwere extracted by3%agarose gel electrophoresis after amplification.5. Using SPSS13.0software processed data.Results:1.The results of FHIT MSP:Using DNA extraction kit to extract DNA of50pairs of lung cancer tissue andnormal lung tissue, then go through MSP and electrophoresis. The positive rate of FHITmethylation was38%(19/50) in lung cancer, while only4%(2/50) in normaltissues, and the difference among them was statistically significant (p=0.000). Therewas no significant difference of FHIT methylation rate between the smoker andnon-smoker(p＞0.05), smoking was independent of lung cancer’s FHIT genemethylation. Gender, age, clinical stage, pathological type of lung cancer and thedifferent histological grade of cancer were independent of lung cancer’s FHIT genemethylation as well, the positive rate was no significant difference between those groups (P>0.05).2.The results of RUNX3MSP:The positive rate of RUNX3gene methylation were24%(12/50) in50cases oflung cancer, while only2%(1/50) in50cases of normal lung tissue which was≥5cmapart from the tumor, and the difference among them was statistically significant (p=0.000). There was no significant difference of RUNX3gene methylation rate betweenthe group of Smokers lung cancer and non-smokers lung cancer (P>0.05).The positiverate of lung cancer in the RUNX3gene methylation was more frequent inadenocarcinoma than that was in squamous cell carcinoma, the difference wasstatistically significant (p <0.05). The RUNX3gene methylation difference was notstatistically significant among gender, age, clinical stage and histological differentiation(P>0.05).3.The results of combining analysis of FHIT and RUNX3MSPThis study studied RUNX3and FHIT genes’ methylation status of patients withlung cancer, and found that there was no relevance of the methylation status in lungcancer specimens between the two genes. FHIT and RUNX3methylation joint test inlung cancer tissues was positive rate higher than the rate of only a single genetic testing.RUNX3and FHIT methylation combined positive rate of smokers was higher than therate of only a single genetic testing, the difference was statistically significant betweenthe smokers with non-smoking group (P <0.05).Conclusions:1, A positive rate of FHIT gene in lung cancer tissues is significantly higher thanthat of normal lung tissue, and there is statistically significant, indicating that the FHITgene methylation asociated with the development of lung cancer. The positive rate ofFHIT gene methylation differences are not statistically significant in terms of gender,age, clinical stage, pathological classification and pathological differentiation amonglung cancer patients.2, the positive rate of RUNX3gene in lung cancer reached24%, while only onecase of normal lung tissue showed methylation positive, the positive rate of the formeris significantly higher than the latter, indicating that RUNX3gene methylation relatedwith the cancer occurrence and development.RUNX3gene methylation inadenocarcinoma is higher than squamous cell carcinoma, but no significant differenceare found between the RUNX3methylation rate and the render, tumor grade andhistological tissues(p＞0.05).3, We found that RUNX3gene and FHIT gene methylation combined test had ahigher positive rate than either a single positive rate of two genes in lung cancer patients,suggesting that methylation detection of multiple genes combined together is more helpfor early lung cancer found.