[Show abstract][Hide abstract]ABSTRACT: The current standard for diagnosis of skin cancers is visual inspection followed by biopsy and histopathology. This process can be invasive, subjective, time consuming, and costly. Optical techniques, including Optical Coherence Tomography (OCT) and Raman Spectroscopy (RS), have been developed to perform non-invasive characterization of skin lesions based on either morphological or biochemical features of disease. The objective of this work is to report a clinical instrument capable of both morphological and biochemical characterization of skin cancers with RS-OCT.
The portable instrument utilizes independent 785 nm RS and 1,310 nm OCT system backbones. The two modalities are integrated in a 4″ (H) × 5″(W) × 8″(L) clinical probe. The probe enables sequential acquisition of co-registered OCT and RS data sets. The axial response of the RS collection in the skin was estimated using scattering phantoms. In addition, RS-OCT data from patients with cancerous and non-cancerous lesions are reported.
The RS-OCT instrument is capable of screening areas as large as 15 mm (transverse) by 2.4 mm (in depth) at up to 8 frames/second with OCT, and identifying locations to perform RS. RS signal is collected from a 44 µm transverse spot through a depth of approximately 530 µm. RS-OCT data sets from a superficial scar and a nodular BCC are reported to demonstrate the clinical potential of the instrument.
The RS-OCT instrument reported here is capable of morphological and biochemical characterization of cancerous skin lesions in a clinical setting. OCT can visualize microstructural irregularities and perform an initial morphological analysis of the lesion. The images can be used to guide acquisition of biochemically specific Raman spectra. The two data sets can then be evaluated with respect to one another to take advantage of the mutually complimentary nature of RS and OCT.
Full-text available · Article · Feb 2011 · Lasers in Surgery and Medicine
[Show abstract][Hide abstract]ABSTRACT: Skin cancer is the most commonly occurring cancer, with incidence rates rising annually. Realizing favorable outcomes requires early diagnosis, for which the current standard is biopsy followed by histopathology. This process can be invasive, subjective, time consuming, and costly. Optical techniques, including Optical Coherence Tomography (OCT) and Raman Spectroscopy (RS), have been developed to perform non-invasive characterization of skin lesions, however neither is without limitation. Here, we demonstrate a clinical instrument for morphological and biochemical characterization of skin cancers with combined RS-OCT. The portable instrument utilizes independent RS and OCT system backbones, and is integrated in a common clinical probe. The potential of the probe for cancer diagnosis is demonstrated on malignant and non-malignant lesions.
Article · Feb 2010 · Proceedings of SPIE - The International Society for Optical Engineering
[Show abstract][Hide abstract]ABSTRACT: Large congenital nevi carry a slightly increased risk of melanoma. Pregnancy poses an additional challenge in the monitoring of these patients because little is known regarding the effects of increased estrogen levels on congenital nevi.
A young woman was observed to have clinical lightening of her garment nevus and satellite nevi during 2 sequential pregnancies. Postpartum, the patient experienced darkening and repigmentation in her large garment nevus, with continued lightening of nearby satellite lesions. In addition to photographic documentation of these changes, biopsy samples taken during pregnant and nonpregnant periods underwent immunohistochemical evaluation for estrogen receptor beta (ERbeta), the predominant estrogen receptor in nevi and melanomas. Biopsy samples collected during pregnancy showed a decrease in nuclear staining for ERbeta compared with samples collected after pregnancy. These changes in ERbeta expression were not associated with histologic atypia during pregnancy or after delivery.
Congenital nevi may be unique in their response to altered estrogen levels. Given the slightly increased risk of melanoma in giant congenital nevi and the dearth of information available regarding the effects of pregnancy on congenital nevi, this case illustrates the need for further study of these pigmented lesions.
[Show abstract][Hide abstract]ABSTRACT: Nonmelanoma skin cancers, including basal cell carcinoma (BCC) and squamous cell carcinoma (SCC), are the most common skin cancers, presenting nearly as many cases as all other cancers combined. The current gold-standard for clinical diagnosis of these lesions is histopathologic examination, an invasive, time-consuming procedure. There is thus considerable interest in developing a real-time, automated, noninvasive tool for nonmelanoma skin cancer diagnosis. In this study, we explored the capability of Raman microspectroscopy to provide differential diagnosis of BCC, SCC, inflamed scar tissue, and normal tissue in vivo.
Based on the results of previous in vitro studies, we developed a portable confocal Raman system with a handheld probe for clinical study. Using this portable system, we measured Raman spectra of 21 suspected nonmelanoma skin cancers in 19 patients with matched normal skin spectra. These spectra were input into nonlinear diagnostic algorithms to predict pathological designation.
All of the BCC (9/9), SCC (4/4), and inflamed scar tissues (8/8) were correctly predicted by the diagnostic algorithm, and 19 out of 21 normal tissues were correctly classified. This translates into a 100% (21/21) sensitivity and 91% (19/21) specificity for abnormality, with a 95% (40/42) overall classification accuracy.
These findings reveal Raman microspectroscopy to be a viable tool for real-time diagnosis and guidance of nonmelanoma skin cancer resection.
Article · Sep 2008 · Lasers in Surgery and Medicine
[Show abstract][Hide abstract]ABSTRACT: Estrogen levels increase during pregnancy and clinical evidence has long suggested that melanocytes are estrogen-responsive. We hypothesized that nevi from pregnant patients would exhibit increased expression of estrogen receptor beta (ERbeta) and thus enhanced potential to respond to altered estrogen levels. Normal, dysplastic and congenital nevi (n = 212) were collected from pregnant and non-pregnant women ranging from 18 to 45 years of age. Immunohistochemical staining was performed on these nevi using antibodies specifically directed against estrogen receptor alpha (ERalpha) and ERbeta. ERalpha was not observed in any lesions; thus, ERbeta was the predominant estrogen receptor in melanocytic cells from all types of nevi. Enhanced positivity for ERbeta in normal nevi during pregnancy was noted, compared with non-pregnant controls including nevocytes residing in both the epidermal and dermal micro-environments (P = 0.005 and P = 0.001 respectively). Nevi with increasingly melanocytic atypia showed increased ERbeta in nevocytes nested within the epidermis. No additional increase in ERbeta in atypical nevi was observed during pregnancy. For normal and congenital nevi, regardless of pregnancy status, dermally associated nevocytes tended to have greater ERbeta immunoreactivity. Significant decreases in ERbeta immunoreactivity were observed in congenital nevi from pregnant women compared with normal and dysplastic nevi from pregnant women. Our data suggest that nevi possess the capacity to be estrogen-responsive. Factors such as pregnancy and degree of atypia are associated with enhanced ERbeta with the exception of congenital nevi where the melanocytes were unique in their response to pregnancy.
[Show abstract][Hide abstract]ABSTRACT: Distinguishing among the various melanocytic proliferations remains one of the biggest challenges in dermatopathology. New insights into tumor proliferation may lead to the discovery of helpful immunohistochemical markers. Akt/PKB is a serine/threonine kinase which is a core component of the PI3K signalling pathway. Activation of Akt is seen in many human cancers. This study evaluated Akt activation (phospho-Akt) by immunohistochemistry on 18 intradermal nevi, 8 dysplastic nevi, 9 Spitz nevi, and 27 melanomas. Immunostaining was graded 0 (no staining), 1 (slightly positive), 2 (moderately positive), and 3 (highly positive). The mean immunostaining scores were as follows: intradermal nevi 1.16, dysplastic nevi 1.25, Spitz nevi 2.11, and melanomas 2.15. Spitz nevi exhibited significant pAkt immunoreactivity. Increased pAkt immunoreactivity is seen in melanomas and Spitz nevi as compared to benign intradermal nevi and dysplastic nevi.
Article · Jun 2008 · Journal of Cutaneous Pathology
[Show abstract][Hide abstract]ABSTRACT: The pleiotropic transcription factor nuclear factor-kappaB (NF-kappaB (p50/p65)) regulates the transcription of genes involved in the modulation of cell proliferation, apoptosis, and oncogenesis. Furthermore, a host of solid and hematopoietic tumor types exhibit constitutive activation of NF-kappaB (Basseres, D. S., and Baldwin, A. S. (2006) 25, 6817-6830). However, the mechanism for this constitutive activation of NF-kappaB has not been elucidated in the tumors. We have previously shown that NF-kappaB-inducing kinase (NIK) protein and its association with Inhibitor of kappaB kinase alphabeta are elevated in melanoma cells compared with their normal counterpart, leading to constitutive activation of NF-kappaB. Moreover, expression of dominant negative NIK blocked this base-line NF-kappaB activity in melanoma cells. Of the three receptors that require NIK for activation of NF-kappaB, only the lymphotoxin-beta receptor (LTbeta-R) is expressed in melanoma. We show in this manuscript that for melanoma there is a strong relationship between expression of the LTbeta-R and constitutive NF-kappaB transcriptional activity. Moreover, we show that activation of the LTbeta-R can drive NF-kappaB activity to regulate gene expression that leads to enhanced cell growth. The inhibition by LTbeta-R shRNA resulted in decreased NF-kappaB promoter activity, decreased growth, and decreased invasiveness as compared with control. These results indicate that the LTbeta-R constitutively induces NF-kappaB activation, and this event may be associated with autonomous growth of melanoma cells.
Full-text available · Article · Jun 2008 · Journal of Biological Chemistry
[Show abstract][Hide abstract]ABSTRACT: We investigate the potential of near-infrared Raman microspectroscopy to differentiate between normal and malignant skin lesions. Thirty-nine skin tissue samples consisting of normal, basal cell carcinoma (BCC), squamous cell carcinoma (SCC), and melanoma from 39 patients were investigated. Raman spectra were recorded at the surface and at 20-microm intervals below the surface for each sample, down to a depth of at least 100 microm. Data reduction algorithms based on the nonlinear maximum representation and discrimination feature (MRDF) and discriminant algorithms using sparse multinomial logistic regression (SMLR) were developed for classification of the Raman spectra relative to histopathology. The tissue Raman spectra were classified into pathological states with a maximal overall sensitivity and specificity for disease of 100%. These results indicate the potential of using Raman microspectroscopy for skin cancer detection and provide a clear rationale for future clinical studies.
[Show abstract][Hide abstract]ABSTRACT: Epithelial cancers, including those of the skin and cervix, are the most common type of cancers in humans. Many recent studies have attempted to use Raman spectroscopy to diagnose these cancers. In this paper, Raman spectral markers related to the temporal and spatial effects of cervical and skin cancers are examined through four separate but related studies. Results from a clinical cervix study show that previous disease has a significant effect on the Raman signatures of the cervix, which allow for near 100% classification for discriminating previous disease versus a true normal. A Raman microspectroscopy study showed that Raman can detect changes due to adjacent regions of dysplasia or HPV that cannot be detected histologically, while a clinical skin study showed that Raman spectra may be detecting malignancy associated changes in tissues surrounding nonmelanoma skin cancers. Finally, results of an organotypic raft culture study provided support for both the skin and the
cervix results. These studies add to the growing body of evidence that optical spectroscopy, in this case Raman spectral markers, can be used to detect subtle temporal and spatial effects in tissue near cancerous sites that go otherwise undetected by conventional histology.
Full-text available · Article · Feb 2008 · Disease markers
[Show abstract][Hide abstract]ABSTRACT: Activated Akt expression (p-Akt) is reportedly increased in many melanomas as compared with benign nevi. The purpose of this study was to evaluate and compare p-Akt immunohistological staining in benign nevi, Spitz nevi and primary melanomas.
Immunostaining for phosphorylated Akt was performed in 41 melanocytic lesions previously classified as benign intradermal nevus (14 lesions), Spitz nevus (9 lesions) or melanoma (18 lesions). Lesions were graded for intensity of p-Akt staining by two independent observers (0, no staining; 1, slightly positive; 2, moderately positive; 3, highly positive). Scores were averaged, and statistical analyses were performed.
Benign nevi showed less staining (mean score 1.18) compared with Spitz nevi (mean score 2.11) and melanomas (mean score 2.19). This difference was statistically significant between benign nevi and melanomas (p = 0.0047) and benign nevi and Spitz nevi (p = 0.0271). No statistical difference was detected in staining between Spitz nevi and melanomas (p = 0.8309).
Activated Akt expression is increased in Spitz nevi and melanomas as compared with benign intradermal nevi, but is unlikely to prove useful in differentiating between the former.
Article · Sep 2007 · Journal of Cutaneous Pathology
[Show abstract][Hide abstract]ABSTRACT: Multiple skin conditions exist which involve clinically significant changes in elastic properties. Early detection of such changes may prove critical in formulating a proper treatment plan. However, most diagnoses still rely primarily on visual inspection followed by biopsy for histological analysis. As a result, there would be considerable clinical benefit if a noninvasive technology to study the skin were available. The primary hypothesis of this work is that skin elasticity may serve as an important method for assisting diagnosis and treatment. Perhaps the most apparent application would be for the differentiation of skin cancers, which are a growing health concern in the United States as total annual cases are now being reported in the millions by the American Cancer Society. In this paper, we use our novel modality independent elastography (MIE) method to perform dermoscopic skin elasticity evaluation. The framework involves applying a lateral stretching to the skin in which dermoscopic images are acquired before and after mechanical excitation. Once collected, an iterative elastographic reconstruction method is used to generate images of tissue elastic properties and is based on a twodimensional (2-D) membrane model framework. Simulation studies are performed that show the effects of three-dimensional data, varying subdermal tissue thickness, and nonlinear large deformations on the framework. In addition, a preliminary in vivo reconstruction is demonstrated. The results are encouraging and indicate good localization with satisfactory degrees of elastic contrast resolution.
Full-text available · Article · Mar 2007 · Proceedings of SPIE - The International Society for Optical Engineering
[Show abstract][Hide abstract]ABSTRACT: Melanomas rarely occur before puberty, have a higher death rate for males, and tend to be more invasive during pregnancy. Prior to the discovery of a second oestrogen receptor (ERbeta), studies with the initial oestrogen receptor, ERalpha, showed no obvious role for oestrogen in the pathophysiology of benign or malignant melanocytic lesions. To investigate the specific immunostaining patterns of ERalpha and ERbeta, benign nevocytic nevi, dysplastic nevi with mild, moderate and severe cytological atypia, lentigo malignas and melanomas of varying depth (Clark) and thickness (Breslow) were studied. ERbeta but not ERalpha was the predominant oestrogen receptor we found in all types of benign and malignant melanocytic lesions. The most intense ERbeta immunostaining was seen in melanocytes in dysplastic nevi with severe cytological atypia and in lentigo malignas. ERbeta expression levels also correlated with the malignant tumor microenvironment; i.e., melanocytes in proximity with keratinocytes>deeper dermal melanocytes in contact with stroma>minimally invasive melanomas>Clark Level III/IV or thick melanomas (Breslow). Discovery that ERbeta expression varies in relation to the tumor microenvironment and increasing depth of invasion suggests its possible usefulness as a surrogate marker for neoplasia and prognosis in malignant melanoma.
[Show abstract][Hide abstract]ABSTRACT: We investigated the reduction of thermal damage to the surrounding tissue when laser incisions were made with and without using thermal conducting templates at room temperature and cooled to 5 degrees C.
We used the Vanderbilt free-electron laser (FEL) at 5.4, 6.1, 6.45, and 7.7 microns. We also used a conventional continuous wave (CW) carbon dioxide laser at 10.6 microns. Incisions were made on 5x10 mm pieces of human breast skin (in vitro) and analyzed with histology. Computer morphometrics were used to measure the amount of thermal damage.
All templates produced a statistically significant reduction in the thermal damage. Additionally, we showed that cooling the templates made a statistically significant greater reduction in the thermal damage. The cooled diamond template reduced the thermal damage from the FEL to 28% of the damage observed without a template. The same cooled template reduced the thermal damage from the CO(2) laser to 56% of the damage observed without a template. Lesser reductions were observed with the copper template and even less with the sapphire template. The sapphire template reduced the thermal damage to 39 and 67% of the damage observed without a template for the FEL and the CO(2) laser, respectively.
These results indicate that unwanted lateral thermal damage from laser incisions can be reduced with cooled thermally conductive templates with the best results obtained with the diamond template, which is also the best thermal conductor.
Article · Oct 2006 · Lasers in Surgery and Medicine
[Show abstract][Hide abstract]ABSTRACT: Paraneoplastic syndromes are systemic reactions in patients with cancers that are unrelated to tumor size or location. Cutaneous paraneoplastic syndromes include proliferative, metabolic, and inflammatory skin disorders. Both systemic and cutaneous paraneoplastic reactions may occur in patients with malignant melanoma. Cancers, including melanoma, may produce growth factors, which may be responsible for proliferative cutaneous paraneoplastic syndromes. A patient with malignant melanoma we previously reported, who had the sudden onset of acanthosis nigricans, skin tags (acrochordons), and seborrheic keratoses provides a model for proliferative cutaneous paraneoplastic syndromes. High levels of α-TGF were found in the patient's urine prior to melanoma removal. The increased level of α-TGF declined after the melanoma was removed, and a corresponding clinical improvement in his acanthosis nigricans, skin tags, and seborrheic keratoses occurred. In the skin lesions, EGF receptors were abnormally present throughout all epidermal layers prior to melanoma removal, and returned to their normal distribution in the basal layers after surgery. Ectopic growth factor production by malignant melanomas and other epithelial neoplasms may cause rare, but distinctive cutaneous paraneoplastic lesions. The model of melanoma, cutaneous paraneoplastic syndromes, and growth factors may provide understanding of both cutaneous lesions associated with neoplasia, and benign cutaneous lesions.
[Show abstract][Hide abstract]ABSTRACT: Previous studies have shown that changing the pulse structure of the free electron laser (FEL) from 1 to 200 ps and thus reducing the peak irradiance of the micropulse by 200 times had little or no effect on both the ablation threshold radiant exposure and the ablated crater depth for a defined radiant exposure. This study focuses on the ablation mechanism at 6.1 and 6.45 microm with an emphasis on the role of the FEL pulse structure. Three different experiments were performed to gain insight into this mechanism. The first was an analysis of the ablation plume dynamics observed for a 1 ps micropulse compared with a 200 ps micropulse as seen through bright-field analysis. Negligible differences are seen in the size, but not the dynamics of ablation, as a result of this imaging. The second experiment was a histological analysis of corneal and dermal tissue to determine whether there is less thermal damage associated with one micropulse duration versus another. No significant difference was seen in the extent of thermal damage on either canine cornea or mouse dermis for the micropulse durations studied at either wavelength. The final set of experiments involved the use of mass spectrometry to determine whether amide bond breakage could occur in the proteins present in tissue as a result of direct absorptions of mid-infrared light into the amide I and amide II absorption bands. This analysis showed that there was no amide bond breakage due to irradiation at 6.45 microm on protein.
Article · May 2005 · Physics in Medicine and Biology
[Show abstract][Hide abstract]ABSTRACT: The Mark-III Free Electron Laser (FEL), tuned to 6.45 microns in
wavelength has been demonstrated to provide for efficient ablation in
ocular, neural, and dermal tissues with minimal collateral damage. To
date, the role of the unique pulse structure of the FEL on the ablation
mechanism has not been determined. In this study, the native pulse
structure of the FEL, a 2.85 gigahertz repetition of picosecond pulses
within a five microsecond macropulse envelope, was changed using a pulse
stretcher. This device changes the duration of the micropulse from its
native one picosecond to 30-200 picoseconds in length, thus reducing the
peak intensity of the micropulse down to 1/200th of the original
intensity, while the macropulse energy remains unchanged. Two basic
metrics were studied: the ablation threshold on water and mouse dermis
and the ablation crater depth on gelatin and mouse dermis. These metrics
were employed at 6.45 and 6.1 microns in wavelength for 1, 100, and 200
picoseconds in micropulse duration. In addition, bright-field imaging
was used to compare the ablation dynamic between 1 ps and 200 ps
micropulses on water at 6.1 and 6.45 microns. The effect of changing the
micropulse duration was also studied on the ablation of mouse dermis for
histological analysis. Craters (500 micron diameter) were created with
25 pulses at three times the ablation threshold as determined for mouse
dermis within 8 hours of removal. Three rows of twenty craters were
created on each piece of mouse dermis for a given parameter set. The
native one picosecond micropulse and 200 picosecond stretched micropulse
were compared at 6.1 and 6.45 microns in wavelength. There was no
difference seen between the native 1 ps micropulse and the stretched
micropulse durations with respect to the ablation threshold, efficiency,
dynamics, and thermal damage.
Article · Apr 2005 · Proceedings of SPIE - The International Society for Optical Engineering
[Show abstract][Hide abstract]ABSTRACT: Several studies have demonstrated Raman spectroscopy to be capable of tissue diagnosis with accuracy rivaling that of histopathologic analysis. This technique obtains biochemical-specific information noninvasively, and can eliminate the pain, time, and cost associated with biopsy and pathological analysis. Furthermore, when used in a confocal arrangement, Raman spectra can be obtained from localized regions of the tissue. Skin cancers are an ideal candidate for this emerging technology, due to their obvious accessibility and presentation at specific depths. However, most commercially available confocal Raman microspectrometers are large, rigid systems ill-suited for clinical application. We developed a bench-top confocal Raman microspectrometer using a portable external-cavity diode laser excitation source. This system was used to study several skin lesions in vitro. Results show the depth-resolved Raman spectra can diagnose in vitro skin lesions with 96% sensitivity, 88% specificity, and 86% pathological classification accuracy. Based on the success of this study, a portable Raman system with a handheld confocal microscope was developed for clinical application. Preliminary in vivo data show several distinct spectral differences between skin pathologies. Diagnostic algorithms are planned for this continuing study to assess the capability of Raman spectroscopy for clinical skin cancer diagnosis.
Article · Jul 2004 · Proceedings of SPIE - The International Society for Optical Engineering
[Show abstract][Hide abstract]ABSTRACT: We have previously shown a reduction in lateral thermal damage with acute studies of skin incisions made in vitro using heat-conducting templates. Here we examined the wound-healing response to laser incisions with heat-conducting templates and explored the use of an optically transparent template with the free electron laser (FEL) at 6.45 microm. First we evaluated the effects of a sapphire heat-conducting template on the lateral thermal damage of FEL incisions using in vitro human skin samples. Next we compared wound tensile strength and histological scoring of the healing of incisions created on the dorsal pelts of live rats with the FEL utilizing metal and sapphire heat-conducting templates and scalpel incisions. The animals were euthanized and the wounds were analyzed at postoperative days 7, 14, and 21. The depth and lateral thermal damage of FEL incisions on in vitro human skin were significantly reduced with the sapphire heat-conducting template. Nonstatistically significant differences in wound tensile strengths and histological scoring of wound healing were noted at days 7 and 14. By day 21, all of the incisions appeared similar. When the data from days 7 and 14 were combined, statistically significant differences were found for each of the templates (except the histological evaluation with the aluminum template) and the scalpel compared with laser incisions made without using a template. The use of metal or sapphire heat-conducting templates reduced the wound-healing delay of laser incisions seen at postoperative days 7 and 14.
[Show abstract][Hide abstract]ABSTRACT: The advantages of the continuous wave (c.w.) CO(2) laser are offset by the delay in laser wound healing secondary to thermal damage. We have developed novel heat-conducting templates to reduce laser thermal damage. Because shortened pulse durations also decrease thermal damage, we tested the effectiveness of heat-conducting templates with a c.w. CO(2) clinical laser and a short-pulsed CO(2) laser to determine the best method and mechanism to minimize thermal damage.
Comparison of 0.2-second shuttered c.w. and 5-microsecond pulsed CO(2) lasers were made by doing incisions on 150 tissue samples from reduction mammoplasties and abdominoplasties. Copper, aluminum, glass, and Plexiglass heat-conducting templates were tested against no template (air) with both lasers. Histological samples were evaluated using computerized morphometrics analysis.
Statistically significant reductions in lateral thermal damage were seen with the copper (50%) and aluminum (39%) templates used with the c.w. CO(2) laser. Only the copper template (39%) significantly reduced thermal damage when used with the pulsed CO(2) laser. Less thermal damage was seen using the pulsed CO(2) laser compared to the c.w. CO(2) laser with each template.
Heat-conducting templates significantly reduced the amount of lateral thermal damage when used with the c.w. CO(2) laser (copper and aluminum) and short-pulsed CO(2) laser (copper). The c.w. CO(2) laser with the copper template compared favorably to the short-pulsed CO(2) laser without a template. Therefore, both heat conductive templates and short-pulse structure provide successful methods for reducing lateral thermal damage, and a combination of the two appears to provide optimal results.
Article · Feb 2003 · Lasers in Surgery and Medicine
[Show abstract][Hide abstract]ABSTRACT: The serine/threonine kinase Akt/protein kinase B and the pleiotropic transcription factor nuclear factor-kappaB [NF-kappaB (p50/p65)] play important roles in the control of cell proliferation, apoptosis, and oncogenesis. Previous studies from our laboratory have shown the constitutive activation of NF-kappaB in melanoma cells. However, the mechanism of this activation is not clearly understood. The purpose of this study was to explore the role of Akt in the activation of NF-kappaB during melanoma tumor progression. Based on our observation that two of the five melanoma cell lines examined exhibit constitutive Akt activation, we evaluated Akt activation by immunohistochemistry in a series of pigmented skin lesions using an antibody specific for phospho-Akt Ser-473. Normal and slightly dysplastic nevi exhibited no significant Akt expression, in marked contrast to the dramatic Akt immunoreactivity seen in severely dysplastic nevi and melanomas (66.3% positive). When these same lesions were stained for nuclear p65, a similar expression pattern was observed. In addition, interruption of Akt activation resulted in increased apoptosis and decreased NF-kappaB promoter activity. These results indicate that activation of Akt kinase is linked to enhanced NF-kappaB nuclear localization and transactivation. We propose that activation of Akt may be an early marker for tumor progression in melanoma.