<i>cir-ITCH</i> involves in the regulation Wnt/b-catenin signaling pathway in vivo.

<p>(A) The linear correlations between the <i>cir-ITCH</i> expression levels and linear <i>ITCH</i> were tested. The relative expression value was normalized by <i>GAPDH</i> expression level. (B) A TCF luciferase reporter assay was performed. The luciferase activity was normalized to the Renilla luciferase activity. (C) The protein levels of Wnt3a and β-catenin was assessed in CRC cells (HCT116 cells and SW480 cells) by Western blot. (D) The mRNA level of <i>c-myc</i> and <i>cyclinD1</i>was detected by quantitative RT-PCR after transfected with <i>cir-ITCH</i> or Control cells in CRC cells. (the upper is <i>c-myc</i> and the lower is <i>cyclinD1</i>) Data are mean±SEM and representative of three independent experiments. (E) HCT116 and SW480 cells were seeded in 96-well plates after been transfected, and cell proliferation was performed daily for 3 days using the CCK-8 assay. Six replicates for each group and the experiment repeated three times. Data are mean±SEM. *<i>P</i><0.05 compared with controls.</p

The sequence of the predicted miRNA binding sites on the the 3'-UTR region of <i>ITCH</i> and <i>cir-ITCH</i>.

<p>The sequence of the predicted miRNA binding sites on the the 3'-UTR region of <i>ITCH</i> and <i>cir-ITCH</i>.</p

The sponges role of <i>cir-ITCH</i> in CRC cells.

<p>(A) Relative luciferase activity of the psiCHECK-2-ITCH constructs co-transfected with miR-20a, miR-7, miR-214 and inhibitor in HCT116 and SW480 cells. In <i>cir-ITCH</i> hyper-expression cells, there were no significant differences in luciferase activity when psiCHECK-2-<i>ITCH</i>-binding site with miRNAs were cotransfected into HCT116 and SW480 cells. Six replicates for each group and the experiment repeated at least three times. Data are mean±SEM. (B) HCT116 and SW480 cells after transfected with <i>cir-ITCH</i> and Control cells were respectively transfected with miR-20a, miR-7 and inhibitor for 24 h and were then further exposed to actinomycin D for 1, 2 and 3 h. Cells were harvested and the stability of <i>cir-ITCH</i> mRNA was analyzed by qRT-PCR relative to time 0 after blocking new RNA synthesis with actinomycin D; data are mean±SEM, normalized to <i>GAPDH</i>.</p

<i>cir-ITCH</i> is correlated with CRC.

<p>(A) Convergent primers can amplify circular RNAs and linear RNAs. Divergent primers amplify circular RNAs only in cDNA compared with genomic DNA (gDNA). GAPDH is linear control. (B) The <i>cir-ITCH</i> was expressed at a higher level in approximately 75.6% of the CRC adjacent tissues compared to match CRC tissues. The expression level of <i>cir-ITCH</i> was analyzed by qRT-PCR based on Taq-man and normalized to <i>GAPDH</i>. Data are represented as mean±SEM from three independent experiments. (C) Random primers and oligodT primers were used respectively in the reverse transcription experiments. The predicted <i>cir-ITCH</i> is absent in poly (A) enriched samples. (D) The predicted <i>cir-ITCH</i> is react against to RNase R treatment. 2-tailed student’s t-test were used in test the differences between groups *p < 0.05 compared to control.</p

Table_2_Exploring the association of addiction-related genetic factors with non-suicidal self-injury in adolescents.xlsx

BackgroundNon-suicidal self-injury (NSSI) is self-injurious behavior without suicidal intent commonly seen in the adolescent population and poses a serious threat to the life safety of adolescents. Related researches suggest a possible correlation between addiction and the occurrence of NSSI. This study aimed to explore the correlation between addiction and NSSI from a molecular biological perspective by analyzing the differential expression of addiction-related genes in NSSI patients.Methods(1) The association between addiction and non-suicidal self-injury in a Chinese adolescent population was verified with the help of questionnaires on substance and non-substance addictions and non-suicidal self-injury among 1,329 adolescents in China, (2) Screening for key genes associated with addiction by bioinformatics analysis, and (3) RT-qPCR experiment was performed to validate key genes and Receiver Operating Characteristic curves were plotted for target genes.Results(1) Substance and non-substance addictions were all significantly correlated with non-suicidal self-injury, (2) Four target genes: SERPINA3, SLC14A1, RPS6 and RPS3A were screened by bioinformatics technique, and (3) Relative quantitative analysis by RT-qPCR revealed that the expression levels of SLC14A1 (p Conclusion(1) The significant association between addiction and NSSI exists in the Chinese adolescent population and (2) Addiction-related genes SLC14A1, RPS6, and RPS3A are differentially expressed in adolescents with NSSI. The genes have the potential to become biological markers for the diagnosis of NSSI.</p

Data_Sheet_1_Exploring the association of addiction-related genetic factors with non-suicidal self-injury in adolescents.docx

BackgroundNon-suicidal self-injury (NSSI) is self-injurious behavior without suicidal intent commonly seen in the adolescent population and poses a serious threat to the life safety of adolescents. Related researches suggest a possible correlation between addiction and the occurrence of NSSI. This study aimed to explore the correlation between addiction and NSSI from a molecular biological perspective by analyzing the differential expression of addiction-related genes in NSSI patients.Methods(1) The association between addiction and non-suicidal self-injury in a Chinese adolescent population was verified with the help of questionnaires on substance and non-substance addictions and non-suicidal self-injury among 1,329 adolescents in China, (2) Screening for key genes associated with addiction by bioinformatics analysis, and (3) RT-qPCR experiment was performed to validate key genes and Receiver Operating Characteristic curves were plotted for target genes.Results(1) Substance and non-substance addictions were all significantly correlated with non-suicidal self-injury, (2) Four target genes: SERPINA3, SLC14A1, RPS6 and RPS3A were screened by bioinformatics technique, and (3) Relative quantitative analysis by RT-qPCR revealed that the expression levels of SLC14A1 (p Conclusion(1) The significant association between addiction and NSSI exists in the Chinese adolescent population and (2) Addiction-related genes SLC14A1, RPS6, and RPS3A are differentially expressed in adolescents with NSSI. The genes have the potential to become biological markers for the diagnosis of NSSI.</p

Image_1_Exploring the association of addiction-related genetic factors with non-suicidal self-injury in adolescents.JPEG

BackgroundNon-suicidal self-injury (NSSI) is self-injurious behavior without suicidal intent commonly seen in the adolescent population and poses a serious threat to the life safety of adolescents. Related researches suggest a possible correlation between addiction and the occurrence of NSSI. This study aimed to explore the correlation between addiction and NSSI from a molecular biological perspective by analyzing the differential expression of addiction-related genes in NSSI patients.Methods(1) The association between addiction and non-suicidal self-injury in a Chinese adolescent population was verified with the help of questionnaires on substance and non-substance addictions and non-suicidal self-injury among 1,329 adolescents in China, (2) Screening for key genes associated with addiction by bioinformatics analysis, and (3) RT-qPCR experiment was performed to validate key genes and Receiver Operating Characteristic curves were plotted for target genes.Results(1) Substance and non-substance addictions were all significantly correlated with non-suicidal self-injury, (2) Four target genes: SERPINA3, SLC14A1, RPS6 and RPS3A were screened by bioinformatics technique, and (3) Relative quantitative analysis by RT-qPCR revealed that the expression levels of SLC14A1 (p Conclusion(1) The significant association between addiction and NSSI exists in the Chinese adolescent population and (2) Addiction-related genes SLC14A1, RPS6, and RPS3A are differentially expressed in adolescents with NSSI. The genes have the potential to become biological markers for the diagnosis of NSSI.</p

Table_1_Exploring the association of addiction-related genetic factors with non-suicidal self-injury in adolescents.xlsx

BackgroundNon-suicidal self-injury (NSSI) is self-injurious behavior without suicidal intent commonly seen in the adolescent population and poses a serious threat to the life safety of adolescents. Related researches suggest a possible correlation between addiction and the occurrence of NSSI. This study aimed to explore the correlation between addiction and NSSI from a molecular biological perspective by analyzing the differential expression of addiction-related genes in NSSI patients.Methods(1) The association between addiction and non-suicidal self-injury in a Chinese adolescent population was verified with the help of questionnaires on substance and non-substance addictions and non-suicidal self-injury among 1,329 adolescents in China, (2) Screening for key genes associated with addiction by bioinformatics analysis, and (3) RT-qPCR experiment was performed to validate key genes and Receiver Operating Characteristic curves were plotted for target genes.Results(1) Substance and non-substance addictions were all significantly correlated with non-suicidal self-injury, (2) Four target genes: SERPINA3, SLC14A1, RPS6 and RPS3A were screened by bioinformatics technique, and (3) Relative quantitative analysis by RT-qPCR revealed that the expression levels of SLC14A1 (p Conclusion(1) The significant association between addiction and NSSI exists in the Chinese adolescent population and (2) Addiction-related genes SLC14A1, RPS6, and RPS3A are differentially expressed in adolescents with NSSI. The genes have the potential to become biological markers for the diagnosis of NSSI.</p

Table_4_Exploring the association of addiction-related genetic factors with non-suicidal self-injury in adolescents.xlsx

BackgroundNon-suicidal self-injury (NSSI) is self-injurious behavior without suicidal intent commonly seen in the adolescent population and poses a serious threat to the life safety of adolescents. Related researches suggest a possible correlation between addiction and the occurrence of NSSI. This study aimed to explore the correlation between addiction and NSSI from a molecular biological perspective by analyzing the differential expression of addiction-related genes in NSSI patients.Methods(1) The association between addiction and non-suicidal self-injury in a Chinese adolescent population was verified with the help of questionnaires on substance and non-substance addictions and non-suicidal self-injury among 1,329 adolescents in China, (2) Screening for key genes associated with addiction by bioinformatics analysis, and (3) RT-qPCR experiment was performed to validate key genes and Receiver Operating Characteristic curves were plotted for target genes.Results(1) Substance and non-substance addictions were all significantly correlated with non-suicidal self-injury, (2) Four target genes: SERPINA3, SLC14A1, RPS6 and RPS3A were screened by bioinformatics technique, and (3) Relative quantitative analysis by RT-qPCR revealed that the expression levels of SLC14A1 (p Conclusion(1) The significant association between addiction and NSSI exists in the Chinese adolescent population and (2) Addiction-related genes SLC14A1, RPS6, and RPS3A are differentially expressed in adolescents with NSSI. The genes have the potential to become biological markers for the diagnosis of NSSI.</p

Table_3_Exploring the association of addiction-related genetic factors with non-suicidal self-injury in adolescents.xlsx

BackgroundNon-suicidal self-injury (NSSI) is self-injurious behavior without suicidal intent commonly seen in the adolescent population and poses a serious threat to the life safety of adolescents. Related researches suggest a possible correlation between addiction and the occurrence of NSSI. This study aimed to explore the correlation between addiction and NSSI from a molecular biological perspective by analyzing the differential expression of addiction-related genes in NSSI patients.Methods(1) The association between addiction and non-suicidal self-injury in a Chinese adolescent population was verified with the help of questionnaires on substance and non-substance addictions and non-suicidal self-injury among 1,329 adolescents in China, (2) Screening for key genes associated with addiction by bioinformatics analysis, and (3) RT-qPCR experiment was performed to validate key genes and Receiver Operating Characteristic curves were plotted for target genes.Results(1) Substance and non-substance addictions were all significantly correlated with non-suicidal self-injury, (2) Four target genes: SERPINA3, SLC14A1, RPS6 and RPS3A were screened by bioinformatics technique, and (3) Relative quantitative analysis by RT-qPCR revealed that the expression levels of SLC14A1 (p Conclusion(1) The significant association between addiction and NSSI exists in the Chinese adolescent population and (2) Addiction-related genes SLC14A1, RPS6, and RPS3A are differentially expressed in adolescents with NSSI. The genes have the potential to become biological markers for the diagnosis of NSSI.</p