Search

Supplementary Data from Loss of SNF5 Expression Correlates with Poor Patient Survival in Melanoma

Purpose: Aberrant expression of SWI/SNF chromatin remodeling complex is involved in cancer development. The tumor suppressor SNF5, the core subunit of SWI/SNF complex, has been shown to regulate cell differentiation, cell cycle control, and apoptosis. To investigate the role of SNF5 in the development of melanoma, we examined the expression of SNF5 in melanocytic lesions at different stages and analyzed the correlation between SNF5 expression and clinicopathologic variables and patient survival.Experimental Design: Using tissue microarry and immunohistochemistry, we evaluated SNF5 staining in 51 dysplastic nevi, 88 primary melanomas, and 48 metastatic melanomas. We studied chemosensitivity of melanoma cells with reduced SNF5 expression by siRNA using cell survival and apoptosis assays.Results: SNF5 expression was reduced in metastatic melanoma compared with dysplastic nevi (P = 0.005), in advanced primary melanoma (Clark's level V) compared with low risk Clark's level II melanoma (P = 0.019), and in melanoma at sun-exposed sites compared with sun-protected sites (P = 0.044). Furthermore, we showed a strong correlation between negative SNF5 expression and a worse 5-year survival in melanoma patients (P = 0.016). Multivariate Cox regression analysis revealed that negative SNF5 expression is an independent prognostic factor to predict patient outcome in primary melanomas (P = 0.031). Finally, we showed that knockdown of SNF5 in melanoma cell lines resulted in significant chemoresistance.Conclusions: Our data indicate that SNF5 may be an important marker for human melanoma progression and prognosis as well as a potential therapeutic target. (Clin Cancer Res 2009;15(20):6404–11)

This study aims to analyze the influence of the Marketing Mix (Product, Price, Place, Promotion, People, Process, and Physical Evidence) on the Consumer Purchasing Decisions. The method used in this study is a quantitative method that is done through the stages of data collection, interpretation of data and appearance of results. Data collection techniques in this study are to use questionnaires. The population in this study was all people domiciled in Kupang City who had used online taxi bike of GrabBike. The technique sampling in this study using Non Probability Sampling with the method of Accidental Sampling, the sample used was 100 respondents .This research uses multiple linear regression analysis. The results show that the Product has a significant influence on the Consumer Purchasing Decisions, the Price has a significant influence on the Consumer Purchasing Decisions, the Place has no influence on the Consumer Purchasing Decisions, the Promotion has no influence on the Consumer Purchasing Decisions, the People has no influence on the Consumer Purchasing Decisions, the Process has a significant influence on the Consumer Purchasing Decisions, and the Physical Evidence has a significant influence on the Consumer Purchasing Decisions. The simultaneous test results in this study revealed that all Marketing Mix variables affect the Consumer Purchasing Decisions. Keywords : Marketing Mix (Product, Price, Place, Promotion, People, Process, Physical Evidence) and Consumer Purchasing Decisions

Objective - Cultural change in the economy has given rise to what might be called 'spiritual entrepreneurship'. The purpose of this paper is to explore the relationship between entrepreneurship and spirituality, even though these two terms are somewhat contradictory. Methodology/Technique - The sample in this study consists of 63 members of the Full Gospel Business Fellowship International (FGBMFI) which is a membership consisting of male businessmen who come from various churches with different business backgrounds. The method used in this study is descriptive analysis. The data was collected using a questionnaire that led to the excavation of information based on previous theoretical studies. The informant selection technique used in this research is accidental. Findings - The results of this study support the findings of previous research in the field of "Spirituality Workplace" and introduces a new model in the field of entrepreneurship. Novelty - Spirituality is a process of human reason in which individual's aim to reach and understand God. On the other hand, entrepreneurs are people who have the courage to take risks to create a business. Hence, in this study, spiritual entrepreneurs refers to people who run their business in line with the values of ethics and religion. Those entrepreneurs interpret their work as worship, honesty and gratitude in addition to usual entrepreneurial behavior (innovation, proactive, competitive aggressiveness, risk taking, autonomy). Type of Paper: Empirical. Keywords: Entrepreneurship; Spirituality; Spiritual Entrepreneurs. JEL Classification: M30, M39.

The minichromosome maintenance (MCM) helicase physically interacts with the recombination proteins Rad51 and Rad52 from yeast to human cells. We show, in Saccharomyces cerevisiae, that these interactions occur within a nuclease-insoluble scaffold enriched in replication/repair factors. Rad51 accumulates in a MCM- and DNA-binding-independent manner and interacts with MCM helicases located outside of the replication origins and forks. MCM, Rad51, and Rad52 accumulate in this scaffold in G1 and are released during the S phase. In the presence of replication-blocking lesions, Cdc7 prevents their release from the scaffold, thus maintaining the interactions. We identify a rad51 mutant that is impaired in its ability to bind to MCM but not to the scaffold. This mutant is proficient in recombination but partially defective in single-stranded DNA (ssDNA) gap filling and replication fork progression through damaged DNA. Therefore, cells accumulate MCM/Rad51/Rad52 complexes at specific nuclear scaffolds in G1 to assist stressed forks through non-recombinogenic functions., This work was supported by grants BFU2015-63698-P and PGC2018-099182-B-I00 (to F.P.) from the Spanish government, and project-ID 393547839-SFB1361 (to H.D.U.) from the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation). M.J.C.-L., M.I.C.-L., C.G.-G, A.Y.-V., and R.G.-P were recipients of pre-doctoral training grants from the Spanish government.

This research was conducted on consumers of PT. Mutiara Timor Star Kupang. The purpose of this study was to determine the effect of promotional strategies and market segmentation partially and simultaneously on purchasing decisions for Honda motorbikes. This research used a qualitative descriptive method approach. The sample data collection used a simple random sampling method. Samples involved in this study were 100 respondents taken from distributed questionnaires, interviews as well as observations to consumers of PT. Mutiara Timor Star Kupang as the primary data source, and reports on Honda motorcycle sales data obtained as secondary data sources. The data analysis technique used in this research was descriptive statistical analysis test, multiple linear regression test, T test, F test, and determination coefficient test using the SPSS version 21 software program. The results of this study indicated that in the paartial approach, the promotional strategy variable did not influence the purchasing decision, while market segmentation variables influenced the purchasing decision. In simultaneous approach, however, the promotional strategy and market segmentation variables influenced the purchasing decisions for Honda motorbikes at PT. Mutiara Timor Star Kupang. Keywords: Promotion Strategy, Market Segmentation, Purchasing Decisions

Penelitian ini bertujuan untuk mengetahui pengaruh gaya kepemimpinan transformasional terhadap kinerja karyawan supermarket Dutalia. Metode yang digunakan dalam penelitian ini adalah metode survei dengan pendekatan kuantitatif asosiatif. Penelitian ini dilakukan di Dutalia Supermarket dengan jumlah sampel 35 orang dengan menggunakan teknik simple random sampling, dan untuk pengumpulan data di lapangan menggunakan kuesioner. Teknik analisis yang digunakan dalam penelitian ini adalah Analisis Regresi Linier Sederhana. Metode uji yang digunakan adalah uji validitas dan uji reliabilitas. Teknik analisis data yang digunakan adalah analisis deskriptif dan analisis inferensial. Hasil penelitian menemukan bahwa secara parsial variabel gaya kepemimpinan transformasional berpengaruh positif dan signifikan terhadap kinerja karyawan di Dutalia Supermarket. Kata Kunci: Gaya Kepemimpinan Tranformasional, Generasi Milenial, Kinerja Karyawan.

The objective of this research is finding the comparison on the raw material stock in Borneo Company by using Economic Order Quantity (EOQ) as the method. The data analysis that been used in this research is Economic Order Quantity (EOQ), the analysis determine the schedule to re-order point and deciding quantity that should be ordered by the company. According to the research’s results which had been conducted in Borneo, The author summarize that Borneo incapable to optimize the calculation toward the raw materials purchased, which the total purchased in Borneo was higher compared to the utilization of the raw materials. As a result this condition lead to the dissipation on total inventory cost. Keywords : Economic Order Quantity (EOQ), Total Inventory Cost (TIC),Safety Stock, and Re-Order Point (ROP)

Dealing with DNA lesions during genome replication is particularly challenging because damaged replication templates interfere with the progression of the replicative DNA polymerases and thereby endanger the stability of the replisome. A variety of mechanisms for the recovery of replication forks exist, but both bacteria and eukaryotic cells also have the option of continuing replication downstream of the lesion, leaving behind a daughter-strand gap in the newly synthesized DNA. In this review, we address the significance of these single-stranded DNA structures as sites of DNA damage sensing and processing at a distance from ongoing genome replication. We describe the factors controlling the emergence of daughter-strand gaps from stalled replication intermediates, the benefits and risks of their expansion and repair via translesion synthesis or recombination-mediated template switching, and the mechanisms by which they activate local as well as global replication stress signals. Our growing understanding of daughter-strand gaps not only identifies them as targets of fundamental genome maintenance mechanisms, but also suggests that proper control over their activities has important practical implications for treatment strategies and resistance mechanisms in cancer therapy.

The purpose of this study is to analyze the level and the type of damage in the production process CV.Bilo Datang and the solutions of quality control are carried out. The method used in this research is quantitative descriptive analysis method using Pareto diagram analysis tools and Cause-effect diagrams. The results of the study showed that the quality control conducted by the company was less than optimal, especially in controlling for half defective products. This is due to several things such as lack of employee skills and maintenance of machinery and equipment that is less than the maximum resulting in the product being half defective. Although in the amount of damage is greater than half defective products but there are also products damaged due to pest attack on wood. The solution provided by the researcher is also appropriate to the stage of the damage that occurred. Keywords: Product Quality Control, Level of damage and Type of damage

Replication of damaged DNA is challenging because lesions in the replication template frequently interfere with an orderly progression of the replisome. In this situation, complete duplication of the genome is ensured by the action of DNA damage bypass pathways effecting either translesion synthesis by specialized, damage-tolerant DNA polymerases or a recombination-like mechanism called template switching (TS). Here we report that budding yeast Pif1, a helicase known to be involved in the resolution of complex DNA structures as well as the maturation of Okazaki fragments during replication, contributes to DNA damage bypass. We show that Pif1 expands regions of single-stranded DNA, so-called daughter-strand gaps, left behind the replication fork as a consequence of replisome re-priming. This function requires interaction with the replication clamp, proliferating cell nuclear antigen, facilitating its recruitment to damage sites, and complements the activity of an exonuclease, Exo1, in the processing of post-replicative daughter-strand gaps in preparation for TS. Our results thus reveal a novel function of a conserved DNA helicase that is known as a key player in genome maintenance.

This study aims to analyze the supporting and inhibiting factors for the development of Oesapa Beach attractions in Kupang City. This type of research is qualitative research using qualitative descriptive research methods. Data sources consist of primary data obtained through observation and interviews; and secondary data sources come from records, interview results and documents from relevant agencies in the study. The results of the study show that the factors that support the development of Oesapa Beach attractions are: (1) Tourist attraction, is anything that has a unique, easy, and value in the form of natural diversity, culture, and man-made results that are targeted or tourist visits; (2) Visitors, are someone who visits a tourist area with the aim of having fun and spending less time, energy and money in less than 24 hours. While the inhibiting factors for the development of Oesapa Beach attractions are: (1) Accessibility, is the degree of ease achieved by a person towards an object, service or environment; (2) Promotion is an effort to offer products or services with the aim of attracting consumers to buy or consume them; (3) Human Resources (HR), namely the workforce involved in tourism is still inadequate and professional; and (4) Funds, is one of the benchmarks for the progress of the development of sustainable tourism objects. Oesapa Beach has a tourist attraction in the form of beautiful and natural scenery, lopo-lopo lined on the beach, and lined with cafes with colorful tents and unique and different cafe designs. In addition to the tourist attraction, visitors who are increasingly busy visiting this tourist attraction become a supporting factor for tourism objects can be developed. Keywords: Analysis, Obstacle Factors, Supporting Factors, Tourism

Penelitian ini dilakukan untuk mengetahui kepuasan pengunjung terhadap fasilitas wisata di Pantai Lasiana. Penelitian ini termasuk jenis penelitian deskriptif. Metode penelitian yang digunakan dalam penelitian ini yaitu metode pendekatan kuantitatif dan kualitatif. Teknik sampel yang digunakan adalah accidential sampling. Sampel yang diambil sebanyak 100 responden yaitu wistawan yang berkunjung ke Pantai Lasiana. Pengumpulan data menggunakan kuisioner (angket), observasi dan studi kepustakaan. Analisis data menggunakan Importance Performance Analysis (IPA). Hasil penelitian menunjukkan tingkat kepuasan konsumen senyatanya lebih rendah dibandingkan dengan tingkat harapan. Hal ini berarti secara keseluruhan pengunjung belum sepenuhnya merasa puas dengan fasilitas wisata yang ada di Pantai Lasiana. Berdasarkan analisis diagram kartesius, fasilitas wisata yang masuk dalam prioritas utama dan memiliki kinerja yang rendah terdiri dari kondisi fisik lopo, kondisi fisik tempat sampah, ketersediaan jumlah tempat sampah dan pemisahan jenis tempat sampah organik dan non organik perlu diperbaiki agar dapat meningkatkan kepuasan wisatawan di masa mendatang.

Polymerase‐blocking DNA lesions are thought to elicit a checkpoint response via accumulation of single‐stranded DNA at stalled replication forks. However, as an alternative to persistent fork stalling, re‐priming downstream of lesions can give rise to daughter‐strand gaps behind replication forks. We show here that the processing of such structures by an exonuclease, Exo1, is required for timely checkpoint activation, which in turn prevents further gap erosion in S phase. This Rad9‐dependent mechanism of damage signaling is distinct from the Mrc1‐dependent, fork‐associated response to replication stress induced by conditions such as nucleotide depletion or replisome‐inherent problems, but reminiscent of replication‐independent checkpoint activation by single‐stranded DNA. Our results indicate that while replisome stalling triggers a checkpoint response directly at the stalled replication fork, the response to replication stress elicited by polymerase‐blocking lesions mainly emanates from Exo1‐processed, postreplicative daughter‐strand gaps, thus offering a mechanistic explanation for the dichotomy between replisome‐ versus template‐induced checkpoint signaling.

DNA lesion bypass is mediated by DNA damage tolerance (DDT) pathways and homologous recombination (HR). The DDT pathways, which involve translesion synthesis and template switching (TS), are activated by the ubiquitylation (ub) of PCNA through components of the RAD6-RAD18 pathway, whereas the HR pathway is independent of RAD18 However, it is unclear how these processes are coordinated within the context of chromatin. Here we show that Bre1, an ubiquitin ligase specific for histone H2B, is recruited to chromatin in a manner coupled to replication of damaged DNA. In the absence of Bre1 or H2Bub, cells exhibit accumulation of unrepaired DNA lesions. Consequently, the damaged forks become unstable and resistant to repair. We provide physical, genetic, and cytological evidence that H2Bub contributes toward both Rad18-dependent TS and replication fork repair by HR. Using an inducible system of DNA damage bypass, we further show that H2Bub is required for the regulation of DDT after genome duplication. We propose that Bre1-H2Bub facilitates fork recovery and gap-filling repair by controlling chromatin dynamics in response to replicative DNA damage.

Tracing DNA repair factors by fluorescence microscopy provides valuable information about how DNA damage processing is orchestrated within cells. Most repair pathways involve single-stranded DNA (ssDNA), making replication protein A (RPA) a hallmark of DNA damage and replication stress. RPA foci emerging during S phase in response to tolerable loads of polymerase-blocking lesions are generally thought to indicate stalled replication intermediates. We now report that in budding yeast they predominantly form far away from sites of ongoing replication, and they do not overlap with any of the repair centers associated with collapsed replication forks or double-strand breaks. Instead, they represent sites of postreplicative DNA damage bypass involving translesion synthesis and homologous recombination. We propose that most RPA and recombination foci induced by polymerase-blocking lesions in the replication template are clusters of repair tracts arising from replication centers by polymerase re-priming and subsequent expansion of daughter-strand gaps over the course of S phase.

Background: Cutaneous melanoma is a life-threatening skin cancer because of its poorly understood invasive nature and high metastatic potential. This study examines the importance of eukaryotic translation initiation factor 5A2 (EIF5A2) in melanoma pathogenesis. Methods: We examined EIF5A2 expression in 459 melanocytic lesions using tissue microarray. In addition, melanoma cell lines were subjected to invasion and cell proliferation assays, zymography, FACS and real-time PCR to investigate the role of EIF5A2 in cancer progression. Results: Positive EIF5A2 staining increased from dysplastic naevi to primary melanomas (PMs; P=0.001), and further increased in metastatic melanomas (P=0.044). Eukaryotic translation initiation factor 5A2 expression was correlated with melanoma thickness (P

Complete and faithful duplication of its entire genetic material is one of the essential prerequisites for a proliferating cell to maintain genome stability. Yet, during replication DNA is particularly vulnerable to insults. On the one hand, lesions in replicating DNA frequently cause a stalling of the replication machinery, as most DNA polymerases cannot cope with defective templates. This situation is aggravated by the fact that strand separation in preparation for DNA synthesis prevents common repair mechanisms relying on strand complementarity, such as base and nucleotide excision repair, from working properly. On the other hand, the replication process itself subjects the DNA to a series of hazardous transformations, ranging from the exposure of single-stranded DNA to topological contortions and the generation of nicks and fragments, which all bear the risk of inducing genomic instability. Dealing with these problems requires rapid and flexible responses, for which posttranslational protein modifications that act independently of protein synthesis are particularly well suited. Hence, it is not surprising that members of the ubiquitin family, particularly ubiquitin itself and SUMO, feature prominently in controlling many of the defensive and restorative measures involved in the protection of DNA during replication. In this review we will discuss the contributions of ubiquitin and SUMO to genome maintenance specifically as they relate to DNA replication. We will consider cases where the modifiers act during regular, i.e., unperturbed stages of replication, such as initiation, fork progression, and termination, but also give an account of their functions in dealing with lesions, replication stalling and fork collapse.

The E3 ligase Rad18 is a key regulator for the lesion bypass pathway, which plays an important role in genomic stability. However, the status of Rad18 expression in melanoma is not known. Using melanoma tissue microarray (TMA), we showed that nuclear Rad18 expression was upregulated in primary and metastatic melanoma compared to dysplastic nevi. Rad18 expression was significantly reduced in sun-exposed sites compared to the sun-protected sites. Strong Rad18 expression correlated with worse 5-year patient survival and was an independent prognostic factor for melanoma found in the sun-protected sites. Furthermore, we showed that melanoma cell proliferation and the expression of pAkt and cyclin D1 were reduced upon Rad18 knockdown. We, for the first time, showed that Rad18 is significantly increased in melanoma and predicts the poor outcome for melanoma in the sun-protected sites. Rad18 is involved in the regulation of melanoma cell proliferation, which can be exploited in designing new strategy for melanoma treatment.

The lesion bypass pathway, which is regulated by monoubiquitination of proliferating cell nuclear antigen (PCNA), is essential for resolving replication stalling due to DNA lesions. This process is important for preventing genomic instability and cancer development. Previously, it was shown that cells deficient in tumour suppressor p33ING1 (ING1b) are hypersensitive to DNA damaging agents via unknown mechanism. In this study, we demonstrated a novel tumour suppressive function of ING1b in preserving genomic stability upon replication stress through regulating PCNA monoubiquitination. We found that ING1b knockdown cells are more sensitive to UV due to defects in recovering from UV-induced replication blockage, leading to enhanced genomic instability. We revealed that ING1b is required for the E3 ligase Rad18-mediated PCNA monoubiquitination in lesion bypass. Interestingly, ING1b-mediated PCNA monoubiquitination is associated with the regulation of histone H4 acetylation. Results indicate that chromatin remodelling contributes to the stabilization of stalled replication fork and to the regulation of PCNA monoubiquitination during lesion bypass.

The Inhibitor of Growth (ING) proteins belong to a well-conserved family which presents in diverse organisms with several structural and functional domains for each protein. The ING family members are found in association with many cellular processes. Thus, the ING family proteins are involved in regulation of gene transcription, DNA repair, tumorigenesis, apoptosis, cellular senescence and cell cycle arrest. The ING proteins have multiple domains that are potentially capable of binding to many partners. It is conceivable, therefore, that such proteins could function similarly within protein complexes. In this case, within this family, each function could be attributed to a specific domain. However, the role of ING domains is not definitively clear. In this review, we summarize recent advances in structure-function relationships in ING proteins. For each domain, we describe the known biological functions and the approaches utilized to identify the functions associated with ING proteins.

The tumour suppressor p33(ING1b) ((ING1b) for inhibitor of growth family, member 1b) is important in cellular stress responses, including cell-cycle arrest, apoptosis, chromatin remodelling and DNA repair; however, its degradation pathway is still unknown. Recently, we showed that genotoxic stress induces p33(ING1b) phosphorylation at Ser 126, and abolishment of Ser 126 phosphorylation markedly shortened its half-life. Therefore, we suggest that Ser 126 phosphorylation modulates the interaction of p33(ING1b) with its degradation machinery, stabilizing this protein. Combining the use of inhibitors of the main degradation pathways in the nucleus (proteasome and calpains), partial isolation of the proteasome complex, and in vitro interaction and degradation assays, we set out to determine the degradation mechanism of p33(ING1b). We found that p33(ING1b) is degraded in the 20S proteasome and that NAD(P)H quinone oxidoreductase 1 (NQO1), an oxidoreductase previously shown to modulate the degradation of p53 in the 20S proteasome, inhibits the degradation of p33(ING1b). Furthermore, ultraviolet irradiation induces p33(ING1b) phosphorylation at Ser 126, which, in turn, facilitates its interaction with NQO1.

Transcription, mRNA decay, translation and protein degradation are essential processes during eukaryotic gene expression, but their relative global contributions to steady-state protein concentrations in multi-cellular eukaryotes are largely unknown. Using measurements of absolute protein and mRNA abundances in cellular lysate from the human Daoy medulloblastoma cell line, we quantitatively evaluate the impact of mRNA concentration and sequence features implicated in translation and protein degradation on protein expression. Sequence features related to translation and protein degradation have an impact similar to that of mRNA abundance, and their combined contribution explains two-thirds of protein abundance variation. mRNA sequence lengths, amino-acid properties, upstream open reading frames and secondary structures in the 5' untranslated region (UTR) were the strongest individual correlates of protein concentrations. In a combined model, characteristics of the coding region and the 3'UTR explained a larger proportion of protein abundance variation than characteristics of the 5'UTR. Further, we used data from human and six other organisms (bacteria, yeast, worm, fly, and plant) and established that steady-state abundances of proteins show significantly higher correlation across these diverse phylogenetic taxa than the abundances of their corresponding mRNAs (p=0.0008, paired Wilcoxon). These data suggest strong selective pressure to maintain protein abundances during evolution, even when mRNA abundances diverge. The absolute protein and mRNA concentration measurements for >1000 human genes and for other organisms represent one of the largest datasets currently available, and reveal both general trends and specific examples of post-transcriptional regulation.

Purpose: Aberrant expression of SWI/SNF chromatin remodeling complex is involved in cancer development. The tumor suppressor SNF5, the core subunit of SWI/SNF complex, has been shown to regulate cell differentiation, cell cycle control, and apoptosis. To investigate the role of SNF5 in the development of melanoma, we examined the expression of SNF5 in melanocytic lesions at different stages and analyzed the correlation between SNF5 expression and clinicopathologic variables and patient survival. Experimental Design: Using tissue microarry and immunohistochemistry, we evaluated SNF5 staining in 51 dysplastic nevi, 88 primary melanomas, and 48 metastatic melanomas. We studied chemosensitivity of melanoma cells with reduced SNF5 expression by siRNA using cell survival and apoptosis assays. Results: SNF5 expression was reduced in metastatic melanoma compared with dysplastic nevi (P = 0.005), in advanced primary melanoma (Clark's level V) compared with low risk Clark's level II melanoma (P = 0.019), and in melanoma at sun-exposed sites compared with sun-protected sites (P = 0.044). Furthermore, we showed a strong correlation between negative SNF5 expression and a worse 5-year survival in melanoma patients (P = 0.016). Multivariate Cox regression analysis revealed that negative SNF5 expression is an independent prognostic factor to predict patient outcome in primary melanomas (P = 0.031). Finally, we showed that knockdown of SNF5 in melanoma cell lines resulted in significant chemoresistance. Conclusions: Our data indicate that SNF5 may be an important marker for human melanoma progression and prognosis as well as a potential therapeutic target. (Clin Cancer Res 2009;15(20):6404–11)

The INhibitor of Growth (ING) genes were discovered during the past decade and identified as type II tumor suppressor genes. Previous studies demonstrated that ING family members participate in various cellular stress responses and thus play important roles in the pathogenesis of various types of cancers, including melanoma. Epidemiological studies showed that UV radiation is the primary etiological factor in melanoma development. Here we review the studies on the role of ING proteins in cellular responses to UV irradiation, melanoma cell motility, and melanoma progression.

Myeloid leukemia-1 (Mcl-1) is an anti-apoptotic protein implicated in tumor progression. Its expression was found to be elevated in many types of human cancers and is correlated with tumor progression. The expression of Mcl-1 in melanoma is not fully understood. We investigated the expression of Mcl-1 in normal nevi, dysplastic nevi, primary melanoma and melanoma metastases by tissue microarray and immunohistochemistry. We found that Mcl-1 expression was significantly increased in dysplastic nevi, primary melanoma and melanoma metastases when compared to normal nevi, though the expression of Mcl-1 was decreased in metastatic melanoma when compared to dysplastic nevi. We did not find any correlation between Mcl-1 expression and melanoma patient survival. Our data suggest that Mcl-1 may play a critical role in the initiation of melanoma development.

Background Trimethylation of the Nε-lysine residues in a protein is one of the most important events of posttranslational modifications. Simple methods for rapid detection and isolation of the Nε-trimethylated protein species are needed. This report introduces a novel method to prepare the affinity purified antibody specific for the Nε-trimethylated lysine (tMeK). The applications of the purified antibody are also reported in this paper. Methods We generated the methylated keyhole limpet heomocyanin (KLH) under controlled chemical methylation reaction using CH3I and used it as an immunogen to raise anti-methylated lysine antibodies. The tMeK specific antibody was selectively isolated using a two-step affinity chromatography in which the mMeK/dMeK specific antibodies were removed and the tMeK specific antibody was captured. Finally, the eluted anti-tMeK antibody was characterized. Results The ELISA results indicated that the antibody reacted only to tMeK but not to mono- and dimethyllysine. Western-blot results showed that the Nε-trimethylated proteins were detected in both animal tissue and cultured cells and that the antibody signal could be competitively inhibited with free tMeK. Conclusion The specific tMeK antibody we developed is useful for one-step isolation of proteins with Nε-trimethyllysine residues and also for the detection, identification and localization of proteins with trimethyllysine residues in the cells.

Melanoma is a life-threatening disease with a high mortality rate due to rapid metastasis. Currently, there is no effective treatment for metastatic melanoma. Integrin-linked kinase (ILK) is a serine/threonine kinase and has its role implicated in connecting cell-extracellular matrix interaction and growth factor signaling to cell survival, cell migration, invasion, anchorage-independent growth, angiogenesis, and epithelial-mesenchymal transition. However, the functional role of ILK in melanoma progression is not completely understood. We have previously shown that strong ILK expression was significantly associated with melanoma thickness. In this study, we further elucidate the role of ILK in melanoma cell migration, invasion, anchorage-independent growth, and tumor growth in vivo by specific ILK knockdown using small interfering RNA and short hairpin RNA. We found that ILK knockdown impeded melanoma cell migration, which was associated with reduced stress fiber formation, cell spreading, and cell adhesion. Furthermore, ILK knockdown decreased the invasion ability of melanoma cells and the formation of anchorage-independent colonies in soft agar. Moreover, ILK knockdown significantly impaired the growth of melanoma xenografts in severe combined immunodeficient mice. This study highlights the importance of ILK in melanoma progression and provides an attractive target for the treatment of melanoma. [Mol Cancer Ther 2007;6(6):1692–1700]