Association Of Genetic Variation Of LncRNA HOTAIR Signaling Pathway With The Susceptibility Of Primary Liver Cancer

ObjectivesPrimary liver cancer?PLC?,referred to as liver cancer,is one of the most common malignant tumors in the world.In recent years,the number of new PLC in China accounts for about half of the total number of new PLC in the world,and PLC patients have poor prognosis and survival rate,resulting in mortality and morbidity that are almost the same.The latest research shows that PLC ranks 4th and 2nd in the new incidence and death spectrum of malignant tumors in Guangdong Province,respectively.Its increasing morbidity and mortality have brought a heavy burden of disease to Guangdong Province,which is a serious threat to residents'health and Malignant tumor of life.PLC is considered to be the result of a combination of environmental exposure and genetic factors.HOX transcript antisense RNA?HOTAIR?,as one of the most researched and most popular LncRNAs,is up-regulated in human malignant tumors and is positively correlated with the occurrence and metastasis of cancer.HOTAIR has complex biological functions.Genetic variation can affect its own structural changes,affect transcription and expression,and thus interfere with the function of downstream target genes.Together with genetic variations of downstream target genes,it can promote the development of PLC.Therefore,this study combines molecular epidemiology,molecular biology,and bioinformatics methods to explore the individual effects of genetic variation in the LncRNA HOTAIR regulatory pathway,the effects of genetic variation-environment,genetic variation interactions and mediation effects on the occurrence of PLC,and further to investigate the effect of genetic variation alone and genetic variation-environment interaction on the prognosis of PLC in HOTAIR regulatory pathways.MethodsA frequency-matched case-control study was used to systematically analyze the genetic variation of the LncRNA HOTAIR regulatory pathway?HOTAIR-EZH2-TP53-PTEN?on susceptibility to PLC in Guangdong population.Comprehensive functional prediction database and bioinformatics tools,combined with domestic and foreign references and tag single nucleotide polymorphism?tag Single Nucleotide Polymorphism,tagSNP?method)to screen genetic variants with potential functions on the HOTAIR regulatory pathway?HOTAIR:rs12427129,rs3816153;EZH2:rs887569,rs12670401,rs80052686,rs6950683;TP53:rs16427129;PTEN:rs2735343?,TaqMan real-time quantitative PCR technology was used to genotype the target genetic variation;Chi-square test was used to compare the differences in environmental factors and genotype distribution between the case group and the control group;MAX method was used to screen the best genetic model for the genetic variation of each gene;single-factor and multi-factor Logistic regression models were used to analyze the influence of environmental factors and genetic variation on the development of PLC;further false discovery rate?False Discovery Rate,FDR?method was used for multiple corrections;haplotype and genetic risk scoring methods were used to analyze the cumulative effect of genetic genetic variation on susceptibility to PLC;interaction terms were introduced in traditional Logistic regression to calculate multiplicative interactions,and Logistic regression was used in combination with Delta formula Calculate additive interactions and explore the interactions between genetic variation,environmental factors,and genetic variation on the development of PLC.Apply multifactor dimensionality reduction?MDR?and classification regression tree?CART?to analyze HOTAIR regulatory pathways.The high-order interaction of mutation-environmental factors affects the susceptibility to PLC.The four-way decomposition method was used to explore the interaction and mediation effects of environmental factors in the association of genetic mutations of various genes with PLC and the contribution rate of each component.Results1.This study included 1453 cases of PLC and 1646 healthy controls.Multivariate Logistic regression analysis showed that chronic HBV infection is associated with the risk of PLC,and the associated OR ratio?OR?and its95%confidence interval?95%CI?were 14.46?95%CI=12.08?17.65?;smoking and drinking were also associated with the risk of PLC?OR=2.76,95%CI=2.22?3.43;OR=2.27,95%CI=1.82?2.82?.2.HOTAIR rs12427129,rs3816153 and PTEN rs2735343 are associated with the risk of PLC in a dominant model.After adjusting for potential confounding factors,individuals carrying the rs12427129T allele reduced the risk of PLC compared with the C allele?OR=0.64,95%CI=0.51?0.80,P<0.001?;compared with individuals carrying the GG genotype,Carrying the rs3816153 GT+TT genotype increases the risk of PLC?OR=1.22,95%CI=1.01?1.46,P=0.039?,and individuals carrying the GC+CC genotype reduce the risk of PLC?OR=0.76,95%CI=0.61?0.94,P=0.012?.Haplotype analysis results showed that HOTAIR TG haplotype carriers reduced the risk of PLC compared with CG haplotype?OR=0.24,95%CI=0.14?0.40,P<0.001?;compared with EZH2 GCCT haplotype,GTCC haplotype carriers reduced the risk of PLC?OR=0.75,95%CI=0.65?0.89,P=0.001?.The results of genetic risk scores showed that the risk of PLC increased with the increase of genetic risk score grades,among which 6-7 risk score grades had the highest risk,which increased the risk of PLC by 3.12 times?OR=3.12,95%CI=1.95?5.00,P<0.001?.3.The association of genetic variation and genetic variation-environmental interaction with PLC showed that HOTAIR rs12427129 had a statistically significant multiplicative interaction with drinking and a multiplicative and additive interaction with chronic HBV infection;HOTAIR rs3816153 has an additive interaction with chronic HBV infection;PTEN rs2735343 has an additive family history with cancer interaction,there is an additive interaction with chronic HBV infection?additional interaction P values are less than 0.05?;there is a multiplicative and additive interaction between HOTAIR rs3816153 and EZH2 rs80052686(P_mul=0.033,P_add=0.045),and EZH2 rs12670401 is a critical statistically significant multiplication and addition interaction(P_mul=0.072,P_add=0.074).4.The MDR method was used to analyze the effects of genetic variation and environmental factors,and high-order interactions between genetic variations on the occurrence of PLC.The MDR results of genetic variation and environmental factors showed the interaction between chronic HBV infection,HOTAIR rs12427129,and rs3816153.The model training samples were accurate.The degree is 0.7939,the accuracy of the verification sample is0.7922,the cross-check consistency is 10/10,and the replacement test is P<0.001;the optimal model of MDR results between genetic variations is EZH2 rs12670401,rs80052686,and PTEN rs2735343,and the corresponding indexes are 0.5568 and 0.5434,respectively,10/10 and P<0.001,suggesting an interaction between the three genetic variations.5.The CART method was used to analyze the effects of genetic variation and environmental factors,and high-order interactions between genetic variations on the occurrence of PLC.The CART analysis of genetic variations and environmental factors showed the interaction of chronic HBV infection,smoking,and PTEN rs2735343;the CART analysis of genetic variations the results showed an interaction between HOTAIR rs12427129,EZH2rs12670401,and PTEN rs2735343.6.Analysis of the mediating effects between genetic variation,environmental factors,and genetic variation using a four-way decomposition method.The results of adjusting mixed factors showed that the effects of HOTAIR rs12427129,rs3816153,and PTEN rs2735343 and chronic HBV infection on PLC were mainly attributed With reference to the interaction effects,the excess relative risks were 0.91?95%CI=0.38?1.45,P=0.001?,-0.30?95%CI=-0.48?-0.11,P=0.002?,and0.52?95%CI=-0.48?-0.11,P=0.037?;the effect of rs2735343 and smoking on liver cancer is mainly due to reference interaction?contribution rate=76.58%,P=0.008?;the effect of rs2735343 and drinking pure mediator on liver cancer has a critical statistical significance?Excess relative risk=0.04,95%CI=-0.00?0.07,P=0.065?.7.EZH2 rs12670401 recessive model?CC vs TT+TC?will increase the risk of portal vein tumor thrombus formation in patients with liver cancer?OR=1.90,95CI=1.42?2.56,P<0.001?.Rs12670401 is associated with smoking and drinking for portal vein tumor thrombus.The formation of multiplication and addition interactions?both P values are less than 0.001?;there is a statistically significant multiplication interaction and critical statistically significant addition interactions between rs12670401 and the family history of cancer for portal vein tumor thrombosis(P_mul=0.005;P_add=0.051).There is a statistically significant multiplication and additive interaction between rs12670401 and smoking for the development of advanced TNM in patients with PLC?both are less than 0.05?;there is a statistically significant multiplication and criticality between rs12670401 and drinking for the development of patients with advanced TNM statistically additive interactions(P_mul=0.034;P_add=0.050).ConclusionsThis study found that genetic variation of HOTAIR regulatory pathways rs12427129,rs3816153,and rs2735343 were closely related to the occurrence of PLC through genetic association analysis.The pairwise interaction between chronic HBV infection and HOTAIR rs12427129,rs3816153 and PTEN rs2735343,drinking and rs12427129,the family history of cancer and rs2735343,can affect the occurrence of PLC.When analyzing the interaction of genetic factors that affect the occurrence of PLC,in addition to discovering that HOTAIR rs3816153 interacts with EZH2 rs12670401 and rs80052686,this study also found high-order interactions between EZH2 rs12670401 and rs80052686 and PTEN rs2735343,HOTAIR rs12427129,and EZH2rs12670401.The complex interactions affect the susceptibility to PLC.Further research found that the genetic variation of EZH2 rs12670401 and its interaction with environmental factors?smoking,drinking,and family history of cancer?can affect the prognosis of PLC?TNM stage and portal vein thrombosis?.The results of this study can provide new ideas and theoretical foundations for the further pathogenesis of PLC,and provide a reference direction for finding potential therapeutic targets for PLC.However,this study found that the association between genetic variation and PLC is limited to the statistical level,and further predicts the function through bioinformatics HOTAIR-EZH2-TP53-PTEN affects the potential biological mechanisms of hepatocarcinogenesis and development.The results still need to be verified by further molecular biological function experiments.