Cellular senescence in usual type uterine leiomyoma Jordan Laser, M.D., Peng Lee, M.D., Ph.D., Jian-Jun Wei, M.D.  Fertility and Sterility  Volume 93, Issue 6, Pages 2020-2026 (April 2010) DOI: 10.1016/j.fertnstert.2008.12.116 Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Photomicrographs illustrate SA-β-Gal stain in primary leiomyoma culture cells and fibroid tissue sections. (A1) Monolayer of leiomyoma cells of 5 days following initiation of the primary culture. (A2) Leiomyoma cells stained by SA-β-Gal following 6 weeks of primary culture with three passages. Blue color is identified as SA-β-Gal positive (see Materials and Methods). (B1–4) Photomicrographs illustrate the various types of tumor cell senescence (SA-β-Gal stain) with a count stain of eosin in a fresh-frozen section of fibroids. (B1) Chimerical type; (B2) patchy and clustered type; (B3) localized type; and (B4) diffuse type. Yellow bars represent 200 μm. Fertility and Sterility 2010 93, 2020-2026DOI: (10.1016/j.fertnstert.2008.12.116) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Senescence analyses of 82 fibroids from 14 symptomatic patients. (A) Pie chart depicting the percentage of fibroids of various levels of senescence. (B) The average percentage of tumor cell senescence (y axis in %) compared with tumor size (x axis in cm). Small t-bars represent standard error. (C) The average percentage of tumor cell senescence (y axis in %) compared with patient's age (x axis). (D) The percentage of tumor cell senescence (x axis in %) compared with tumor cell proliferation index (y axis in % of immunopositivity for Ki-67). A moderate inverse correlation (r = −.59) was observed. Fertility and Sterility 2010 93, 2020-2026DOI: (10.1016/j.fertnstert.2008.12.116) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Correlation analysis of let-7 expression in association with the levels of tumor cell senescence. (A) Photonegative image illustrates the intensity of cDNA products of let-7c, d and f-2 obtained by RT-PCR and visualized in 1.5% agarose gel. RNA from seven nonsenescent (left, Non-SN) and seven highly senescent (right, High-SN) fibroids were examined. The three let-7 miRNAs and control U6 were listed. (B) Quantitative analysis of the relative abundance of let-7 miRNA products in highly senescent (gray bars) and nonsenescent (black bars). Significant difference of let-7 expression was observed in all three let-7 family members (P<.05). (C) Photomicrographs illustrate the intensity of let-7c expression in highly senescent (High-SN, left) and nonsenescent (Non-SN, right) fibroids. (D) Correlation analysis of let-7c expression (detected by MicroRNA in situ hybridization, in y axis) and percentage of tumor cell senescence (x axis) in 63 fibroids (see Results). A moderate correlation (r = .44) was observed. Fertility and Sterility 2010 93, 2020-2026DOI: (10.1016/j.fertnstert.2008.12.116) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions