The Association Study Between Environmental Factors,Long Non-coding RNAs,and The Risk Of Hepatocellular Carcinoma In Male Population

[Objective]To explore risk factors and protective factors of hepatocellular carcinoma(HCC)among male population of Xianen.and to investigate the IncRNA expression profile between cancer tissues and adjacent tissues collected from male HBV-associated HCC cases.in addition to that,we conducted a meta-analysis to evaluate the diagnostic performance of IncRNA in HCC.[Materials and Methods]1.We employed a 1:1 matched case-control study and conduct a unified questionnaire interview along 264 male HCC cases and same number of control subjects.Univariate and multivariate conditional regressions were applied to calculate the odds ratio and 95%confident interval of-all included factors.Random forest was used to identify the major factors which related with the HCC onset on the male population of Xiamen.Moreover,stratified analysis was also conducted by different HBsAs status.2.The microarray chip was used to identify and screen the differentially expressed RNAs between cancer tissue and adjacent tissue collected from 10 male HBV-associated HCC cases,and qRT-PCR validation.After the normalization of signal intensity.the fold change of each RNA was calculated.We applied following criteria to identify differentially expressed RNAs which are absolute fold change greater 2,a P value less than 0.05 and have more than 3 biological replicas.The clustering analysis was used to investigate the distance on expression profile between samples,and Cytoscape software was used to establish lncRNA-1 RNA co-expression network.We also conducted GO and pathxvay enrichment analysis in differentially expressed mRNAs.3.We conducted a literature search on the studies about lncRNAs and their diagnostic performance in HCC by following meta-analysis protocol.After literature inclusion and evaluation,data extraction was performed.After that,heterogeneity estimation,effect size synthesis,evaluation of publication bias and clinical application were conducted.[Results]1.The multivariate regression showed that HBsAg positive.consumption of smoked food,s1oking and passive smoking,history of liver diseases are the risk factors of HCC,while the consumption of fruit and tea have protective effect against HCC risk.The random forest suggested that HBsAg positive and dietary habit have major impact on the onset of HCC among Xiamen male population.2.The statistical analysis identified that in total 1517 lncRNAs and 2826 1RNAs were differentially expressed between cancer tissues and adjacent tissues.The GO and pathway enrichment analysis conducted in differentially expressed mRNAs showed that they are closely associated vvith the absorption.metabolism and response of metallic ions.3.In total,12 eligible studies were included in the meta-analysis,the effect size synthesis showed that the sensitivity was 0.86.specificity was 0.86.positive likelihood ratio was 6.4,negative likelihood ratio was 0.16,and the diagnostic odds ratio was 39.02.The SROC showed that the area under curve was 0.93.The result of linear regression indicated that there was no sign of obvious publication bias.Overall,the meta-analvsis suggested that lncRNA has high diagnostic performance in HCC.[Conclusion]1.HBsAg positive,smoking and passive smoking,consumption of smoked food were identified as the risk factors of HCC,and random forest suggest that the HBsAg positive and dietary habit have major impact on the onset of HCC.However.the data suggested that the proportion of adopting unhealthy lifestyle among HBsAg positive subjects was relatively high.Therefore,it requires intervention and the promotion of regular medical examination.2.The microarray analysis identified several differentially expressed lncRNAs,and after validation on both independent samples and large population,they may have the potential of serving as the biomarkers of diagcnosis and prognosis.The enrichment analysis showed that differentially expressed nRNAs are closely related with the metallic ion,indicating that the exposure of heavy metal mayrelated with the HCC onset among Xiamen male population.3.The meta-analysis demonstrated that the IncRNA has high diagnostic performance in HCC.and even better than conventional serum biomarkers.However,it still requires further studies conducted in large population to validate.