Screening And Analysis Of Differential Expressed LncRNAs In Lung Cancer In Yunnan XuanWei,China

ObjectiveIn our country, Yunnan Xuanwei is the high incidence of lung cancer, give priority to the non-small cell lung cancer. Have data shows:in Yunnan Xuanwei, the average mortality rate is 98.10/10 million of male lung cancer, and 83.28/10 million of female lung cancer.The occurrence of lung cancer is usually higher than rural city, while in Yunnan Xuanwei, non-smoking female lung cancer mortality rate is also high, can reached 120/10 million, and the highest can reach 400/10 million, is 20 times, other parts of China leads the world. The main reason with Xuanwei high lung cancer mortality is the late stage of cancer patients whlie diagnosed and tumor cell invasion and metastasis have occurred, the lack of effective treatment measures. Therefore, the study of xuanwei lung cancer development mechanism is particularly important.This topic first analyze the structure, function and mechanism of LncRNAs, then apply LncRNAs chip of Yunnan Xuanwei lung cancer tissue filtered differentially expressed LncRNAs analysis, then the application of real time PCR technology to verify the results of gene chip screening, the final target genes of projections for screening of LncRNAs and validation, understand its role in the yunnan xuanwei lung cancer development, in order to further understand Xuanwei the pathogenesis of lung cancer, and provide a scientific basis for early diagnosis and treatment.Methods1. Using LncRNAs microarrays （AGI_LncRNAs+mRNA₄×180k） to screen the differentially expressed LncRNAs of 5 fresh specimens Yunnan Xuanwei lung cancer tissue and paracancerous tissue. Fold change≥2 are acceptable, then the differentially expressed LncRNAs were obtained.2. The expression of the key LncRNAs between cancer tissue and paracancerous tissue were valitaed using RT-PCR; And explore the relationship of its expression level between cancer tissue and paracancerous tissue;3. Statistical analysis was performed on the clinical data of patients, gender, age, smoking, lymph node metastasis, clinical stage and histological type of the relationship between the expression of LncRNAs in patients with different level and the key molecular study screened.Result1. Using the microarray results of Yunnan Xuanwei Lung cancer as the basis, in Yunnan Xuanwei Lung cancer we obtained in 1544 abnormal expression LncRNAs. According to the fold difference is greater than or equal to 2 and the original signal value is greater than or equal to 50, we obtained in 367 abnormal expression genes, 133 of the LncRNAs is up-regulated,234 of the LncRNAs down-regulated;2. Combined with the literature analysis, we find out the 6 key LncRNAs has the research value. The up regulation of expression of the 3, down regulated 3, they are: SGOL1-AS1、SNHG4、PVT1、FENDRR、H19. PWRN1;3. Using real-time fluorescence quantitative PCR （real-time quantitative PCR） technology to detect the expression of 6 LncRNAs screened in Yunnan Xuanwei lung cancer tissue and paracancerous tissue, The results found that SGOL1-AS1、SNHG4 and PVTlwere up-regulated, FENDRR、H19 and PWRN1 were down-regulated in Xuanwei Lung cancer tissue;4. There is no significant correlation between the SGOL1-AS1、SNHG4、PVT1、 FENDRR、H19, but PWRN1 and tumor size has a certain correlation.Conclusions1. Using LncRNAs microarray found a large number of up-regulated or down regulated expression of LncRNAs exists in Yunnan Xuanwei Lung cancer tissues;2. Use real-time PCR technology to verify the six LncRNAs, the results and the microarray results with the same direction. For the further study of osteosarcoma Yunnan Xuanwei Lung cancer provides clues.3. Statistical analysis of clinical data, results show that the six LncRNAs expression level has no correlation with the patient’s gender, age, smoking, lymph node metastasis and histological type.While the expression of PWRN1 difference level related to the patients with tumor size.