human zr 75 1  (ATCC)

Journal: Journal of Cell Communication and Signaling

Article Title: Epigenetic regulation of breast ductal carcinoma in situ by miR‐217 through DNMT1 and Hedgehog‐GLI pathway

doi: 10.1002/ccs3.70030

Figure Lengend Snippet: Screening of DCIS‐related genes and prediction of upstream regulatory factors. (A) Venn diagram of differentially methylated genes in promoter regions predicted by the R packages methylKit (left) and eDMR (right); overlapping region indicates shared genes; (B) volcano plot of the GSE21422 microarray for DCIS‐related expression (black: nonsignificant; green: significantly downregulated; red: significantly upregulated); (C) prediction of hypermethylated and low‐expressed genes in DCIS promoter regions (left: top 520 downregulated genes in GSE21422 ; right: top 100 hypermethylated genes predicted by both methylKit and eDMR); (D) Expression of TSHZ2 in BC samples recorded in the GEPIA database (red: tumor; gray: normal); (E) prediction of CpG islands in the TSHZ2 promoter region by MethPrimer (blue regions represent CpG islands); (F) expression of the DNMT1 gene in the GSE21422 microarray for DCIS‐related expression; (G) prediction of upstream regulatory miRNAs of DNMT1 (blue: miRNAs predicted by mirDIP; red: significantly downregulated miRNAs in BC from GSE45666 ; overlapping area: common candidates). BC, breast cancer; DCIS, ductal carcinoma in situ; DNMT1, DNA methyltransferase 1; TSHZ2, teashirt zinc finger homeobox 2.

Article Snippet: The human DCIS cell line ZR‐75‐1 (ATCC® CRL‐1500TM) was purchased from the American Type Culture Collection.

Techniques: Methylation, Microarray, Expressing, In Situ

Journal: Journal of Cell Communication and Signaling

Article Title: Epigenetic regulation of breast ductal carcinoma in situ by miR‐217 through DNMT1 and Hedgehog‐GLI pathway

doi: 10.1002/ccs3.70030

Figure Lengend Snippet: Validation of miR‐217 targeting DNMT1 in ZR‐75‐1 cells. (A) Target binding and mutation sites of miR‐217 and DNMT1. (B) Dual‐luciferase reporter assay validating the targeting relationship between miR‐217 and DNMT1; (C, D) RT‐qPCR analysis of miR‐217 and DNMT1 expression in clinical samples. (E, F) RT‐qPCR analysis of miR‐217 and DNMT1 expression in different cell groups. (G) Correlation analysis of miR‐217 and DNMT1 expression in DCIS samples. Data are presented as mean ± standard deviation, and cell experiments were repeated three times. DCIS, ductal carcinoma in situ; DNMT1, DNA methyltransferase 1; NC, negative control. * indicates p < 0.05 compared to the mimic‐NC or Controlgroup; # indicates p < 0.05 compared to the inhibitor‐NC group.

Article Snippet: The human DCIS cell line ZR‐75‐1 (ATCC® CRL‐1500TM) was purchased from the American Type Culture Collection.

Techniques: Biomarker Discovery, Binding Assay, Mutagenesis, Luciferase, Reporter Assay, Quantitative RT-PCR, Expressing, Standard Deviation, In Situ, Negative Control

Journal: Journal of Cell Communication and Signaling

Article Title: Epigenetic regulation of breast ductal carcinoma in situ by miR‐217 through DNMT1 and Hedgehog‐GLI pathway

doi: 10.1002/ccs3.70030

Figure Lengend Snippet: Effect of miR‐217 targeting DNMT1 on DCIS cell proliferation, migration, and invasion. (A) RT‐qPCR analysis of miR‐217 expression in different groups of ZR‐75‐1 cells. (B) Western blot analysis of DNMT1 protein expression in different groups of ZR‐75‐1 cells. (C) CCK‐8 assay for cell proliferation. (D) Monoclonal formation assay for cell clonogenic capacity. (E, F) Transwell assays for cell migration and invasion. Data are presented as mean ± standard deviation, and cell experiments were repeated three times. DCIS, ductal carcinoma in situ; DNMT1, DNA methyltransferase 1; NC, negative control. * indicates p < 0.05 compared to themimic‐NC + oe‐NC group; # indicates p < 0.05 compared to the miR‐217‐mimic + oe‐NC group.

Article Snippet: The human DCIS cell line ZR‐75‐1 (ATCC® CRL‐1500TM) was purchased from the American Type Culture Collection.

Techniques: Migration, Quantitative RT-PCR, Expressing, Western Blot, CCK-8 Assay, Tube Formation Assay, Standard Deviation, In Situ, Negative Control

Journal: Journal of Cell Communication and Signaling

Article Title: Epigenetic regulation of breast ductal carcinoma in situ by miR‐217 through DNMT1 and Hedgehog‐GLI pathway

doi: 10.1002/ccs3.70030

Figure Lengend Snippet: Effect of DNMT1 regulation on TSHZ2 expression and DCIS cell proliferation, migration, and invasion. (A) RT‐qPCR and Western blot analysis of TSHZ2 expression in clinical samples (N: adjacent normal tissues; T: tumor tissues). (B) RT‐qPCR and Western blot analysis of TSHZ2 expression in ZR‐75‐1 cells after DNMT1 expression modulation. (C) RT‐qPCR and Western blot analysis of TSHZ2 expression in ZR‐75‐1 cells after miR‐217 expression modulation; (D) RT‐qPCR and Western blot analysis of TSHZ2 expression in ZR‐75‐1 cells after combined modulation of miR‐217 and DNMT1 expression. (E) Prediction of CpG islands using MethPrimer and MSP analysis of TSHZ2 promoter region methylation. (F) ChIP assay assessing DNMT1 enrichment at the TSHZ2 promoter region. (G) CCK‐8 assay for cell proliferation; (H) Monoclonal formation assay for clonogenic capacity. (I, J) Transwell assay for cell migration and invasion. Data are presented as mean ± standard deviation, and cell experiments were repeated three times. ChIP, chromatin immunoprecipitation; DCIS, ductal carcinoma in situ; DNMT1, DNA methyltransferase 1; MSP, methylation‐specific PCR; NC, negative control; TSHZ2, teashirt zinc finger homeobox 2. * indicates p < 0.05 comparedto the Control group, oe‐NC group, mimic‐NC group, or oe‐NC + mimic‐NC group; # indicates p < 0.05 compared to the si‐NCgroup, inhibitor‐NC group, or inhibitor‐NC + oe‐NC group; & indicates p < 0.05 compared to the miR‐217‐inhibitor + oe‐NC group or oe‐DNMT1 group; @ indicates p < 0.05 compared to the si‐DNMT1group.

Article Snippet: The human DCIS cell line ZR‐75‐1 (ATCC® CRL‐1500TM) was purchased from the American Type Culture Collection.

Techniques: Expressing, Migration, Quantitative RT-PCR, Western Blot, Methylation, CCK-8 Assay, Tube Formation Assay, Transwell Assay, Standard Deviation, Chromatin Immunoprecipitation, In Situ, Negative Control, Control

Journal: Journal of Cell Communication and Signaling

Article Title: Epigenetic regulation of breast ductal carcinoma in situ by miR‐217 through DNMT1 and Hedgehog‐GLI pathway

doi: 10.1002/ccs3.70030

Figure Lengend Snippet: Effect of TSHZ2 regulation on the Hedgehog‐GLI signaling pathway and DCIS cell proliferation, migration, and invasion. (A) RT‐qPCR and Western blot analysis of TSHZ2 transfection efficiency in ZR‐75‐1 cells after TSHZ2 expression modulation. (B, C) RT‐qPCR and Western blot analysis of Hedgehog‐GLI pathway‐related factors (GLI1 and SHH) in different cell groups. (D) CCK‐8 assay for evaluating cell proliferation. (E) Monoclonal formation assay for assessing clonogenic capacity; (F, G) Transwell assay for measuring cell migration and invasion. Data are presented as mean ± standard deviation, and cell experiments were repeated three times. DCIS, ductal carcinoma in situ; NC, negative control; SHH, Sonic Hedgehog; TSHZ2, teashirt zinc finger homeobox 2. * indicates p < 0.05 compared to the oe‐NC group; # indicates p < 0.05 compared to the si‐NC group; & indicates p < 0.05 compared to the oe‐TSHZ2 + DMSO group; @ indicates p < 0.05compared to the si‐TSHZ2 + DMSO group.

Article Snippet: The human DCIS cell line ZR‐75‐1 (ATCC® CRL‐1500TM) was purchased from the American Type Culture Collection.

Techniques: Migration, Quantitative RT-PCR, Western Blot, Transfection, Expressing, CCK-8 Assay, Tube Formation Assay, Transwell Assay, Standard Deviation, In Situ, Negative Control

Journal: Journal of Cell Communication and Signaling

Article Title: Epigenetic regulation of breast ductal carcinoma in situ by miR‐217 through DNMT1 and Hedgehog‐GLI pathway

doi: 10.1002/ccs3.70030

Figure Lengend Snippet: Effect of miR‐217 on DNMT1/TSHZ2/Hedgehog‐GLI axis and tumor formation in DCIS cells in vivo. (A) Representative images of tumors from each group of nude mice. (B) Comparison of tumor volumes among different groups of nude mice. (C) Comparison of tumor masses among different groups of nude mice; (D, E) RT‐qPCR and Western blot analysis of miR‐217, DNMT1, TSHZ2, GLI1, and SHH expression in tumor tissues from different groups of nude mice. (F) MSP analysis of TSHZ2 promoter region methylation and ChIP analysis of DNMT1 enrichment at the TSHZ2 promoter region. (G) TUNEL staining for measuring apoptosis in tumor tissues from different groups of nude mice. Data are presented as mean ± standard deviation. n = 6. ChIP, chromatin immunoprecipitation; DCIS, ductal carcinoma in situ; DNMT1, DNA methyltransferase 1; MSP, methylation‐specific PCR; NC, negative control; SHH, Sonic Hedgehog; TSHZ2, teashirt zinc finger homeobox 2. * indicates p < 0.05 compared to theLv‐NC + oe‐NC group; # indicates p < 0.05 compared to the Lv‐miR‐217 + oe‐NC group.

Article Snippet: The human DCIS cell line ZR‐75‐1 (ATCC® CRL‐1500TM) was purchased from the American Type Culture Collection.

Techniques: In Vivo, Comparison, Quantitative RT-PCR, Western Blot, Expressing, Methylation, TUNEL Assay, Staining, Standard Deviation, Chromatin Immunoprecipitation, In Situ, Negative Control