Berberine inhibits the proliferation of human uterine leiomyoma cells

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Berberine inhibits the proliferation of human uterine leiomyoma cells Hsiao-Li Wu, M.S., M.B.A., Tung-Yueh Chuang, Ph.D., Ayman Al-Hendy, M.D., Ph.D., Michael P. Diamond, M.D., Ricardo Azziz, M.D., M.B.A., M.P.H., Yen-Hao Chen, Ph.D. Fertility and Sterility Volume 103, Issue 4, Pages 1098-1106 (April 2015) DOI: 10.1016/j.fertnstert.2015.01.010 Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

Figure 1 BBR blocks the E2- and P4-induced proliferation of UtLM cells. UtLM cells were incubated with BBR and/or E2 (A) or P4 (B) at the indicated concentrations for 72 hours. Cell viability was determined by the MTS assay. The cell survival rate of untreated cells was considered as 100%. After ANOVA analysis, data were separated into three different groups (a, b, and c). The differences among the three groups were significant (P<.01). No significant differences were found between treatments inside one group. Data are shown as means ± SE; n = 3. Fertility and Sterility 2015 103, 1098-1106DOI: (10.1016/j.fertnstert.2015.01.010) Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

Figure 2 BBR inhibits cell proliferation and induces cell cycle arrest in UtLM and UtSMC cells. UtLM and UtSMC cells were incubated with BBR at the indicated concentrations for 72 and 24 hours for cell proliferation assay and cell cycle test. (A) Bright field pictures (×100) were taken after treatment. (B) Cell viability was determined by MTS assay; the cell survival rate of untreated cells was considered as 100%. (C) The gene expression of the proliferation markers, MKI67 and PCNA, after BBR treatment is seen. (D) Cells were fixed and stained with PI and analyzed by a flow-cytometer. Quantitation of the PI staining data is presented as the cell cycle distribution percentages. (E) Changes in the expression of cyclin A1 and cyclin B1 with BBR treatment. (F) Changes in the expression of p21 and p53 with BBR treatment. (G) Changes in the expression of CDK1 and CDK4 with BBR treatment. Data are shown as means ± SE; n = 3. ∗P<.05 and ∗∗P<.01 vs. 0 μM of BBR. Fertility and Sterility 2015 103, 1098-1106DOI: (10.1016/j.fertnstert.2015.01.010) Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

Figure 2 BBR inhibits cell proliferation and induces cell cycle arrest in UtLM and UtSMC cells. UtLM and UtSMC cells were incubated with BBR at the indicated concentrations for 72 and 24 hours for cell proliferation assay and cell cycle test. (A) Bright field pictures (×100) were taken after treatment. (B) Cell viability was determined by MTS assay; the cell survival rate of untreated cells was considered as 100%. (C) The gene expression of the proliferation markers, MKI67 and PCNA, after BBR treatment is seen. (D) Cells were fixed and stained with PI and analyzed by a flow-cytometer. Quantitation of the PI staining data is presented as the cell cycle distribution percentages. (E) Changes in the expression of cyclin A1 and cyclin B1 with BBR treatment. (F) Changes in the expression of p21 and p53 with BBR treatment. (G) Changes in the expression of CDK1 and CDK4 with BBR treatment. Data are shown as means ± SE; n = 3. ∗P<.05 and ∗∗P<.01 vs. 0 μM of BBR. Fertility and Sterility 2015 103, 1098-1106DOI: (10.1016/j.fertnstert.2015.01.010) Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

Figure 3 BBR down-regulates PTTG-1, E2F1, and COX-2 expression in UtLM and UtSMC cells. UtLM and UtSMC cells were incubated with BBR at the indicated concentrations for 24 hours. Gene expression was determined by real-time RT-PCR. Data are shown as means ± SE; n = 3. ∗P<.05 and ∗∗P<.01 vs. 0 μM of BBR. Fertility and Sterility 2015 103, 1098-1106DOI: (10.1016/j.fertnstert.2015.01.010) Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

Figure 4 BBR induces apoptosis in UtLM and UtSMC cells. UtLM and UtSMC cells were incubated with BBR at the indicated concentrations for 24 hours. (A) Annexin V staining of UtLM cells (×100). (B) Changes in BAX gene expression with BBR treatment. Data are shown as means ± SE; n = 3. ∗∗P<.01 vs. 0 μM of BBR. Fertility and Sterility 2015 103, 1098-1106DOI: (10.1016/j.fertnstert.2015.01.010) Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions