Selective Expression of FLIP in Malignant Melanocytic Skin Lesions

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Presentation transcript:

Selective Expression of FLIP in Malignant Melanocytic Skin Lesions Roberto R. Bullani, Bertrand Huard, Isabelle Viard-Leveugle, Jean-Hilaire Saurat, Lars E. French Journal of Investigative Dermatology Volume 117, Issue 2, Pages 360-364 (August 2001) DOI: 10.1046/j.0022-202x.2001.01418.x Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 The AL129 antibody specifically recognizes FLIP protein.(A) Immunoblot analysis of lysates from FLIP- (left lane) and mock-transfected Jurkat cells using the rabbit antihuman FLIP antibody AL129. (B) Sensitivity of Jurkat and Jurkat-FLIP cells to recombinant FasL and TRAIL. Journal of Investigative Dermatology 2001 117, 360-364DOI: (10.1046/j.0022-202x.2001.01418.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Selective expression of FLIP in melanomas. (A–C) Immunohistochemical staining with AL129 anti-FLIP antibody of an atypical nevus (A), a superficial spreading melanoma (B), and a nodular melanoma (C), showing increased FLIP expression in malignant melanocytic skin lesions (B and C) as compared with benign ones (A). (D) Immunohistochemical staining of a nodular melanoma with an isotype-matched antibody. Scale bar: 100 μm. Journal of Investigative Dermatology 2001 117, 360-364DOI: (10.1046/j.0022-202x.2001.01418.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Semiquantitative analysis of FLIP expression in 61 benign and malignant human melanocytic skin lesions. Semiquantitative evaluation of the FLIP immunohistochemical staining intensity in the melanocytic component of common benign melanocytic nevi (CBMN), atypical nevi (AN), superficial spreading melanomas (SSM), nodular melanomas (NM), and cutaneous melanoma metastases (CMM). 0, no staining; +, weak staining; ++, moderate staining; +++, strong staining. Similar results were obtained using another rabbit antihuman FLIP antibody (AL148). Journal of Investigative Dermatology 2001 117, 360-364DOI: (10.1046/j.0022-202x.2001.01418.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 The percentage of cells expressing FLIP within melanocytic lesions increases with melanoma thickness. Percentage of FLIP-positive cells with respect to the total number of melanocytes within melanocytic lesions were analyzed by morphometry. CBMN, common benign melanocytic nevi; AN, atypical nevi; SSM, superficial spreading melanoma; NM, nodular melanoma; CMM, cutaneous melanoma metastases. Journal of Investigative Dermatology 2001 117, 360-364DOI: (10.1046/j.0022-202x.2001.01418.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 FLIP transfection enhances the resistance of melanoma cells to TRAIL and FasL. The TRAIL and FasL-sensitive melanoma cell line WK was transfected with an empty (WK-mock) or with a FLIP-containing expression vector (WK-FLIP). RT-PCR analysis of FLIP expression in FLIP-transfectants (A). Sensitivity (% survival) of established clones to TRAIL and FasL was tested after 16 h incubation with 100 ng per ml of recombinant human TRAIL (B) or FasL (C), respectively. Jurkat cells served as positive controls for TRAIL and FasL. Asterisks (*) denote FLIP-transfected clones for which the difference in sensitivity to TRAIL or FasL, as compared with the mock-transfected cell line, achieved statistical significance (t test, p <0.01). Each clone was tested in quintuplicate, and experiments were performed three times. Results of one representative experiment are shown. Journal of Investigative Dermatology 2001 117, 360-364DOI: (10.1046/j.0022-202x.2001.01418.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions