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Figure 3 | Breast Cancer Research

Figure 3

From: 17beta-hydroxysteroid dehydrogenase type 1 modulates breast cancer protein profile and impacts cell migration

Figure 3

mRNA and protein level modulation by 17beta-hydroxysteroid dehydrogenase type 1 (17β-HSD1). (A) Expression of 17β-HSD1, proliferating cell nuclear antigen (PCNA), nm23-H1, and BRCA2 and CDKN1A interacting protein (BCCIP) in wild type (WT) MCF7 cells and MCF7 cells stably transfected with 17β-HSD1 (MCF7-17βHSD1) revealed by western blots. β-actin protein amount was used as internal control. The arrows show the positions of the protein bands. (B) The western blot bands in (A) were quantified, and the ratio between the protein signals of interest and the β-actin signal was calculated to determine the relative protein expression values for WT MCF7 and MCF7-17βHSD1. (C) Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) values (mRNA copies/µg total RNA) of mRNAs encoding proteins involved in estradiol production and/or action in WT MCF7 and MCF7-17βHSD1. N, negligible (RT-qPCR values < 1,000); 0, mRNA not detected after many rounds of amplification. (D) Relative mRNA expression values of enzymes involved in estradiol production in MCF7-17βHSD1 as compared to WT MCF7. The mRNA levels in WT MCF7 cells were fixed at 100. (E) and (F) Relative 17β-HSD1 (E) and nm23-H1 (F) mRNA expression in siRNA-transfected T47D cells. T47D cells were transfected with 17β-HSD1 siRNA or control siRNA and 17β-HSD1 mRNA was quantified by RT-qPCR. mRNA quantity in control-siRNA transfected cells was fixed at 100. Error bars represent standard deviation. *P <0.05 analyzed by Student's t-test.

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