Article Figures & Data
Figures
- Figure 1.
Correlation of expression of progestagen-associated endometrial protein (PAEP) with microphthalmia-associated transcription factor (MITF) in human melanoma tissues. The gene expression profiles of 16 primary (MIS, melanoma in situ; thin, < 1 mm; IM, intermediate melanoma, 1-4 mm; thick, > 4 mm) and 40 metastatic melanoma (MM) specimens were examined utilizing an Affymetrix Human Genome U133 Plus 2.0 Array platform. Normalized signal intensity values for PAEP (206859_s_at) and MITF (207233_s_at) genes are analyzed by MAS 5.0 analysis software and the calculated Pearson correlation coefficient (r) is 0.86 (P < .05).
- Figure 2.
Correlation of expression of progestagen-associated endometrial protein (PAEP) with microphthalmia-associated transcription factor (MITF) in melanoma daughter cells. Tumor cells isolated from freshly excised thick primary (A) and metastatic melanoma samples (B) were passaged and expanded in vitro (C). (D) The expression of PAEP and MITF genes was examined by semi-qRT-PCR in 10 melanoma daughter cell lines (No. 3-12), 3 National Cancer Institute (NCI) melanoma cell lines (No. 13-15), 1 human normal skin sample (No. 1) and 1 normal human epidermal melanocyte (NHEM; No. 2). (E) The densitometry of specific bands for PAEP and MITF genes was analyzed according to β-actin with AlphaEase FC image analysis software. 1. skin; 2. NHEM, thick primaries; 3. MCC13; 4. MCC80A, lymph node metastases; 5. MCC67; 6. MCC74; 7. MCC80B, distant metastases; 8. MCC12A; 9. MCC12F; 10. MCC69A; 11. MCC69B; 12. MCC81, NCI metastatic cell lines; 13. 624-Mel; 14. 624.38-Mel; 15. A375. The calculated Pearson correlation coefficient (r) of these 2 genes was 0.75 (P = .001).
- Figure 3.
Transfection of MCC69B and 624.38-Mel metastatic melanoma cells with microphthalmia-associated transcription factor (MITF) small interfering RNA (siRNA) specifically diminishes progestagen-associated endometrial protein (PAEP) gene expression. (A) Four duplex siRNA; siMITF5, 6, 7, and 8; and a pool of siMITF5-8 were separately transfected into melanoma cells. MITF expression was examined by real-time qRT-PCR at 48 h after transfection. The expression of MITF and PAEP genes was examined by real-time qRT-PCR (B) and Western blotting (C) after siMITF5-8 transfection in melanoma cells at 48 h and 72 h after transfection, respectively. The expression of PAEP and MITF genes was examined by real-time qRT-PCR (D) and Western blotting (E) after siPAEP10-12 transfection in melanoma cells. Mock (wild-type) and nonspecific siControl transfection served as negative controls.
- Figure 4.
Silencing of microphthalmia-associated transcription factor (MITF) or progestagen-associated endometrial protein (PAEP) gene expression decreases the migration of MCC69B and 624.38-Mel metastatic melanoma cells. 624.38-Mel cells with or without transfection of small interfering (si) PAEP10-12 or siControl (A), or with or without transfection of siMITF5-8 or siControl (B), were plated onto the upper transwell chambers and allowed to migrate for 6 h. 624.38-Mel (C) or MCC69B (D) cells that migrated to the underside of the chamber were counted in 10 randomly chosen fields under a microscope and were analyzed by Dunnett t-test, with the relative migration ratio of wild-type cells set as 100% (P < .05).
