The Analysis Of Breast Cancer Serum MicroRNAs Expression Spectrum Variance And Functional Verification

Objective:we choose breast cancer patients and normal physical examination personnel as the research objects to observe the difference of the expression profile of miRNAs in serum, and establish the differential expression profile of miRNA. To speed up the application of serum miRNA as a biomarker to clinical transformation and Provide new potential biomarkers for the differential diagnosis.At the same time, the preliminary study on the function of miRNA in breast cancer has laid a foundation for the study of the mechanism and function of miRNA in the development of breast cancer.MethodsWe selected 10 cases of breast cancer patients and 10 normal persons and MiRNA sequencing of 10 breast cancer tissues, adjacent tissues, serum and normal human serum was performed by high throughput sequencing. And the specific expression of miRNAs in breast cancer was screened out by using Clip-seq, TargetScan and other databases. Using real-time fluorescence quantitative RT-PCR to screen the results of large samples of serum verification, screening for the differential expression of miRNA,.The cell viability of inhibitor miRNA was detected by CCK-8, the target gene was predicted, which laid the foundation for the functional study.ResultsFirstly, miRNA high-throughput sequencing result of pathological tissue and breast cancer tissues and normal human serum, Through the comparison of different samples of data Clean, the analysis of the common Reads and the specific Reads, Fold (Change log2)|>=1, P-value<0.01 for the differential expression of the selected standard, Differential Expressions of miRNAs were selected in 986 different expressions,.Secondly, using realtime RT-PCR miRNA to verify the results of sequencing screening. CT RT-PCR value With the differential expression of more than 2 times and less than 30 as the standard.4 miRNA were screened out. That is, hsa-miR-374a-5p、hsa-miR-223-3p、hsa-miR-423-5p and Validation analysis of serological expression of hsa-miR-320;Then, in 224 cases of breast cancer patients and 132 healthy control group, the expression of miRNA in serum was quantified by quantitative analysis. The results showed that the relative expression of hsa-miR-374a-5p, hsa-miR-223-3p, hsa-miR-423-5p and in the control group and breast cancer group were up-regulated in miRNA,, and hsa-miR-320 in serum, The difference was statistically significant (P<0.05). The expression of miRNA in serum of 4 patients with breast cancer was consistent with the expression of the early stage breast cancer.Finally, we selected the sequencing results and verified the miRNA, using CCK8 to detect the cells after transfection. MiR-223-3p and miR-423-5p have the function to regulate the activity of breast cancer cells through the target gene prediction and functional classification, lay the foundation for the next research on loss of functionConclusion:In this topic, we used a new generation of high throughput sequencing to screen miRNA in breast cancer and normal serum. The differential expression profile of miRNA in breast cancer was initially established. Serological validation of the four miRNA large samples, which were up-regulated in the sequencing results, was considered the expression trends of hsa-miR-374a-5p, hsa-miR-223-3p, hsa-miR-423-5p, hsa-miR-320 in serum and the expression of the breast cancer tissues in the breast cancer tissues were correlated.It is the Potential biomarkers for the diagnosis of breast cancer, Hsa-miR-423-5p, hsa-miR-423-5p in breast cancer cells have a functional expression, can provide direction for its functional research.