| 1.Mechanism of Interleukin-33 Regulating Tumor Stem Cell Activity in Hepatocellular CarcinomaObjective Interleukin-33(interleukin-33,IL-33),as a pro-inflammatory factor,is involved in inflammation,allergic diseases,tissue repair,and tumors by binding its receptor carcinogenicity inhibitory gene 2(Suppression of Tumorigenicity 2,ST2)And other physiological processes,but their role in liver cancer is not yet clear.This study intends to use The Cancer Genome Atlas(TCGA)database to analyze the differential expression of IL-33 mRNA in hepatocellular carcinoma and its adjacent tissues,the correlation of clinicopathological parameters and its potential regulatory genes;The effect of cellular tumor stemness.This will provide a new medical basis and theoretical basis for the diagnosis and treatment of liver cancer.Methods 1.Expression and prognosis analysis of IL-33 in hepatocellular carcinoma Download the expression profiling sequencing data and clinical data of hepatocellular carcinoma from the TCGA database(https://cancergenome.nih.gov/)in the Gene Expression Omnibus(GEO)database,and use R software and TCGA database for a comprehensive integration analysis,The difference of IL-33 m RNA expression between normal cancer tissues and liver cancer tissues was analyzed.The correlation between IL-33 and the survival time of liver cancer was analyzed by Kaplan-Meier curve and single factor Cox.At the same time,gene expression enrichment analysis(GSEA)method was used to analyze the Kyoto Encyclopedia of Genes and Genomes(KEGG)database to predict IL-33 regulatory genes.2.Study on the effect of interleukin-33 on the activity of hepatocellular carcinoma tumor stem cells In vitro ball formation experiments are used to determine whether a single liver cancer cell has the ability to renew itself in a suitable conditioned medium.Real-time quantitative PCR method was used to detect the expression of tumor stem cell genes of liver cancer cells treated with IL-33 and other treatments,to explore the effect of IL-33 on genes related to liver cancer stem cell activity.Using 5-fluorouracil(5-Fu)chemotherapeutic drugs and CCK8 experiment to evaluate the effect of IL-33 on drug resistance of liver cancer cells.Results 1.Expression and prognosis analysis of IL-33 in epatocellular carcinoma We found that the expression of IL-33 m RNA in liver cancer tissues was lower than that in normal tissues(p<0.05),and the survival time of liver cancer patients with high expression of IL-33 was longer than that of low expression groups(p <0.05).Single factor Cox analysis found 33 is a protective factor for liver cancer.The higher the expression level,the longer the survival time of liver cancer(HR=0.796);but multi-factor Cox analysis found that IL-33 is not an independent prognostic factor for liver cancer.IL-33 may participate in the occurrence and development of liver cancer through TGF-β signaling pathway,JAK/STAT signaling pathway and the corresponding receptor interaction signaling pathway.2.Mechanism of interleukin-33 regulating liver cancer stem cell activity(1)IL-33 inhibits the ability of hepatoma cells to form balls in vitro The human hepatoma cells Hep G2,HUH-7,SMMC-7721 were stimulated with different concentrations of human recombinant IL-33.Under the stimulation of 5 ng / m L human recombinant IL-33,the number of balls formed was significantly reduced compared to the control group,and as the concentration increased,the number of hepatoma cells formed into spheres decreased(p <0.05).(2)IL-33 inhibits the expression of tumor stem genes in liver cancer cells Under the stimulation of IL-33,the expression of the dry genes NANOG,NOTCH3,SOX2,OCT-3,NOTCH1,and NOTCH2 of human hepatoma cells Hep G2,HUH-7,SMMC-7721 was down-regulated(p <0.05).(3)IL-33 inhibits the resistance of hepatocellular carcinomacells to 5-Fu Hepatocellular carcinoma(Hep G2,HUH-7,SMMC-7721)stimulated with IL-33(25ng / ml),relative to the control group,the experimental group at 5-Fu concentration of 50μg / ml the drug resistance decreased significantly(p<0.05).Conclusions 1.IL-33 m RNA was under-expressed in hepatocellular carcinoma,IL-33 was a protective factor for patients with hepatocellular carcinoma,but not an independent prognostic factor for patients with hepatocellular carcinoma;the expression of IL-33 has a role in the progression of hepatocellular carcinoma Inhibition.2.IL-33 may inhibit the activity of tumor stem cells of hepatocellular carcinomaand thus inhibit the occurrence and development of hepatocellular carcinoma.2.Single nucleotide polymorphisms of CD44,IL-33 and ST2 and susceptibility to hepatocellular carcinomaObjective To explore the relationship between the single nucleotide polymorphisms(SNPs)of IL-33/ST2 and tumor stem cell marker CD44 gene and the risk factors and clinical related indicators of susceptibility to hepatocellular carcinoma(HCC).Methods Using case-control research methods,565 patients with liver cancer and 561 normal medical examiners in our hospital were included.The SNa Pshot method is used for genotyping selected genes.The population genetics Hardy-Weinberg(HWE)law was used to test whether the included case and control populations were from the same Mendelian population.Unconditional Logistic regression was used to analyze confounding factors such as gender,age and calculate relative risk factors(odds ratio,OR)and its 95% confidence interval(CI)to analyze alleles of the selected gene locus And genotype frequency and the risk of HCC.Chi-square test was used to analyze whether the distribution of selected gene polymorphisms and single nucleotide polymorphisms in the sub-groups of clinicopathological characteristics were different.Results Through analysis,it was found that the polymorphisms of the CD44 rs187115 and ST2 rs3821204 GG sites were associated with an increased risk of developing liver cancer(p <0.05),and there was no correlation between the polymorphism of the rs1929992 site of the IL-33 gene and the risk of HCC.Conclusions CD44 gene rs187115 and ST2 gene rs3821204 may increase the risk of hepatocellular carcinoma;... |
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