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Fig. 4 | Journal of Hematology & Oncology

Fig. 4

From: The long noncoding RNA H19 promotes tamoxifen resistance in breast cancer via autophagy

Fig. 4

Beclin1 is regulated by H19 lncRNA via the H19/SAHH/DNMT3B axis. a The effects of H19 knockdown on the mRNA expression levels of autophagy-related genes in MCF7/TAMR cells were analyzed by qRT-PCR. The data are presented as the mean ± SD of three independent experiments. Student’s t test was used for statistical analysis. b The mRNA expression of Beclin1 in MCF7/TAMR cells overexpressing H19 or the control vector was evaluated by qRT-PCR. The data are presented as the mean ± SD of three independent experiments. Student’s t test was used for statistical analysis. c The expression of Beclin1 in MCF7/TAMR cells with knockdown or overexpression of H19 was analyzed by Western blotting. The value next to each blot is the quantification of the relative expression of the indicated band normalized to GAPDH expression. d DNA methylation in the promoter region of Beclin1 in MCF7/TAMR cells that stably expressed shControl (shCtrl) or shH19 was determined by real-time quantitative methylation-specific polymerase chain reaction (QMSP). The data are presented as the mean ± SD of three independent experiments. Student’s t test was used for statistical analysis. e The expression of SAHH, Beclin1, and LC3 in MCF7/TAMR cells with single H19 knockdown or double knockdown of H19/SAHH was analyzed by Western blotting. The value next to each blot is the quantification of the relative expression of the indicated band normalized to GAPDH expression. f The expression of DNMT3B, Beclin1, and LC3 in MCF7/TAMR cells with single H19 knockdown or double knockdown of H19/DNMT3B was analyzed by Western blotting. The value next to each blot is the quantification of the relative expression of the indicated band normalized to GAPDH expression. g DNA methylation in the promoter region of Beclin1 in MCF7/TAMR cells that stably expressed shCtrl, shH19, shH19+shSAHH, or shH19+shDNMT3B was determined by QMSP. The data are presented as the mean ± SD of three independent experiments. Student’s t test was used for statistical analysis. h The binding of DNMT3B to the promoter region of the Beclin1 gene was determined by a ChIP assay. The input was used as a positive control, and normal rabbit IgG was used as a negative control. The data are presented as the mean ± SD of three independent experiments. Student’s t-test was used for statistical analysis. i mRNA expression of Beclin1 in 37 breast cancer tissue samples. The data are presented as the mean ± SD; n = 37. The Wilcoxon signed-rank test was used for statistical analysis. k Spearman correlation analysis of H19 and Beclin1 expression in 37 breast cancer tissue samples. Spearman correlation coefficients and P values were calculated. *P < 0.05, **P < 0.01 compared with the control group. n.s. indicates no significant difference

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