| Research Background and PurposeHepatocellular carcinoma?HCC?is one of the most common malignancies worldwide.To further explore the relationship between new gene function changes and the development and malignant features of liver cancer.It is of great significance to reveal the precise molecular mechanism of its occurrence and development,to design rational therapeutic drugs and to judge its prognosis,and to further improve the treatment level of liver cancer in China.Micro RNAs RNAs?mi RNAs?are non-coding single stranded RNAs.It plays an important role in many physiological processes,such as development,cell proliferation,apoptosis and hormone secretion.By binding to oncogenes or tumor suppressor genes,it regulates a variety of tumorigenesis processes,including cell proliferation,angiogenesis,differentiation,and apoptosis,and drug resistance.This study constructed,validated and evaluated signature mi RNAs and biological pathway analysis based on large sample mature mi RNAs downloaded from the TCGA database.There are two main parts.Part I:?1?Screening differentially expressed genes of mi RNAs and m RNAs in HCC and paracancer tissues;?2?Basing on statistical analysis,build Risk scores related to iconic mi RNAs and conduct random grouping verification;?4?Analyzing the regulatory mechanism network of iconic mi RNAs.Part II:?1?The expression of marker mi RNAs was verified in HCC and paracancer tissues;?2?The signature mi RNAs were verified at the cellular level to investigate their effect on the apoptosis of Hep G2 cells.Methods?1?The data analyzed in the study were downloaded from TCGA.Screening differential genes of mi RNAs and m RNAs.Samples with complete survival information and differential mi RNA expression profiles were randomly divided into two groups?Train group and Test group?.Signal mi RNAs were obtained by single factor and multi-factor Cox regression analysis base on mi RNAs of Train group.According to the product of each iconic mi RNA and the sum of its coefficients,a prognostic Risk Score containing multiple mi RNA was established.Risk Score=??1×expression level of mi RNA1?+??2×expression level of mi RNA2?+??3×expression level of mi RNA3?+...+??n×expression level of mi RNAn?.Risk Score was verified in the whole group,Train group and Test group.The top 10 core genes were identified according to Cytoscape 3.6.1 and its plug-ins.?2?The expression levels of Signal mi RNAs were verified in HCC tissues and adjacent matched normal tissues by real-time fluorescent quantitative PCR.The effect of different expression of mi RNAs on apoptosis of Hep G2 cells was observed by flow cytometry.Results?1?In the Train group,multivariate Cox risk regression results showed that hsa-mir-101-3p?HR:0.747,95%CI:0.567-0.985?,hsa-mir-139-5p?HR:0.727,95%CI:0.591-0.884?and hsa-mir-516a-5p?HR:1.123,95%CI:1.028-1.227?were independent prognos tic factors for HCC patients?P<0.05?.?2?The overall survival rate with high expression of hsa-mir-101-3p and high expression of hsa-mir-139-5p was higher than low expression of mi RNAs in HCC patients?P<0.05?.The overall survival rate with low expression of hsa-mir-516a-5p was higher than that high expression of hsa-mir-516a-5p in HCC patients?P<0.05?.?3?The overall survival rate of Hgih risk group in the whole group,Train group and Test group was lower than that of the corresponding Low risk group?P<0.05?.According to ROC curve analysis,the areas of the three ROC curves were>and 0.7,and the differences were statistically significant?P<0.05?.The correlation model between Risk Score and survival and death of HCC patients indicated that the model had clinical significance?P<0.05?.?4?Mir-101-3p and mir-139-5p were highly expressed in paracancer tissues and poorly expressed in cancer tissues,but mir-516a-5p was poorly expressed in paracancer tissues and highly expressed in cancer tissues.?5?Overexpression of mir-101-3p and mir-139-5p promoted apoptosis of Hep G2cells.Low expression of mir-516a-5p promoted apoptosis of Hep G2 cells.Conclusion?1?Mir-101-3p and mir-139-5p were positively correlated with the prognosis of HCC patients.Mir-516a-5p was negatively correlated with the prognosis of HCC patients.?2?The construction of relevant Risk score based on mir-101-3p,mir-139-5p,and mir-516a-5p can distinguish patients with poor prognosis from those with HCC without being affected by clinical variables.?3?mir-101-3p and mir-139-5p were tumor suppressor genes in HCC patients.mir-516a-5p is a oncogenic gene in HCC patients. |
PDF Full Text Request