Hair follicles (HFs) undergo life-long cyclical transformations, progressing through stages of rapid growth (anagen), regression (catagen), and relative “quiescence” (telogen). Since HF cycling abnormalities underlie many human hair growth disorders, the accurate classification of individual cycle stages within skin biopsies is clinically important and essential for hair research. For preclinical human hair research purposes, human scalp skin can be xenografted onto immunocompromised mice to study human HF cycling and manipulate long-lasting anagen in vivo. While available for mice, a comprehensive guide on how to recognize different human hair cycle stages in vivo is lacking. Here, we present such a guide, which uses objective, well-defined, and reproducible criteria and integrates simple morphological indicators with advanced, (immuno)-histochemical markers. This guide also characterizes human HF cycling in xenografts and highlights the utility of this model for in vivo hair research. Detailed schematic drawings and representative micrographs provide examples of how best to identify human HF stages, even in sub-optimally sectioned tissue, and practical recommendations are given for designing human-on-mouse hair cycle experiments. Thus, this guide seeks to offer a benchmark for human hair cycle stage classification, for both hair research experts and newcomers to the field.

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To identify candidate genes that could be used as diagnostic and therapeutic targets for hepatocellular carcinoma (HCC), we searched for the genes that are overexpressed in HCC by combining representational difference analysis and microarray. Genes such as glypican-3 (GPC3), insulin-like growth factor 2, long-chain fatty-acid-coenzyme A ligase 4, farnesyl diphosphate synthase were frequently identified in our screening. Northern blot analysis with these four genes confirmed their overexpression in HCC. Among them we found that GPC3 transcript is upregulated in six out of seven cases of HCC. Immunoblot and immunohistochemical staining using polyclonal anti-GPC3 antibodies further confirmed that GPC3 protein is indeed increased in HCC tumor samples. We also found that GPC3 is secreted into culture media from cell lines derived from HCC. We conclude that GPC3 is a good molecular marker for HCC. (Cancer Sci 2003; 94: 259-262) lypican-3 (GPC3) is a member of the glypican family of heparan-sulfate proteoglycans, which are linked to the cell surface through a glycosylphosphatidylinositol anchor.1) GPC3 loss-of-function mutation in human causes type 1 Simpson-Golabi-Behmel syndrome (SGBS1), an X-linked condition characterized by pre-and postnatal overgrowth.2) GPC3 knockout mice indeed exhibited several phenotypic features of SGBS1. [3][4][5] These findings together with cell line-specific promotion of apoptosis by OCI-5/GPC3 6) suggest that GPC3 plays a negative role in cell proliferation and an apoptosis-inducing role in specific tissues.Consistent with the above idea, GPC3 expression is frequently silenced by promoter methylation in ovarian cancer cell lines, 7) rat mesothelioma cell lines and human primary tumors, 8) and breast cancer cell lines. 9) In addition, ectopic expression of GPC3 inhibited growth in some of the above cell lines, suggesting a tumor-suppressive role of GPC3. In contrast, GPC3 is known to be overexpressed in hepatocellular carcinoma, 10,11) neuroblastoma and Wilms' tumor cells.12) The role of GPC3 in these tumors is not known. It is also not known whether GPC3 protein is indeed increased in these tumors.Hepatocellular carcinoma (HCC) is one of the most common tumors worldwide and is one of the leading causes of death among cancer patients in Korea. Identification of genes that are overexpressed in HCC not only helps our understanding of tumorigenesis, but also helps to develop diagnostic and therapeutic targets. In this study, we combined representational difference analysis (RDA) 13) and microarray 14) to identify genes that are frequently overexpressed in HCC tumor samples. Since GPC3 was the most frequently obtained gene in our screening, we further evaluated it as a tumor marker for HCC. Materials and MethodsTumor samples and cell lines. HCC tumor tissues and corresponding normal liver tissues were obtained from patients (Table 1) undergoing surgery in Kyungpook National University Hospital (Daegu, Korea) with the approval of the human research review committee and the patients' consent. C...

nesterified arachidonic acid (also called free AA) entering cells exogenously or released endogenously is rapidly converted to AA-CoA esters by the catalytic action of fatty acidCoA ligase (FACL), particularly by the AA-preferring FACL4.1, 2) Several lines of evidence indicated that the level of free AA in cells regulates apoptosis.3-7) More recently, it has been demonstrated that free AA induces apoptosis by activating the caspase-3 pathway, and that the induction of apoptosis by inhibitors of AA metabolism is a consequence of its accumulation.8) Moreover, overexpression of FACL4 prevented AA-induced apoptosis by reducing the level of intracellular free AA. 8)These findings, together with overexpression of FACL4 in colon adenocarcinoma, strongly suggest that the FACL4 pathway may be important in colon carcinogenesis.9) In addition, cyclooxygenase-2 (Cox-2), another AA-utilizing enzyme, is also known to be overexpressed in colon cancer.10, 11) Therefore, AA metabolism plays an important role in colon carcinogenesis. Currently, overexpression of FACL4 and Cox-2 in colon adenocarcinoma is believed to deplete free AA, thereby removing a proapoptotic signal and promoting carcinogenesis. 8,9) Hepatocellular carcinoma (HCC) is one of the most common tumors worldwide and is one of the leading causes of death among cancer patients in Korea. Identification of genes that are overexpressed in HCC is of importance to understand liver tumorigenesis and to develop diagnostic and therapeutic targets. As is seen in colon cancer, Cox-2 is known to be overexpressed in human HCC, [12][13][14] implying that AA metabolism might also be important in liver carcinogenesis. However, it is not known whether FACL4 is also overexpressed in HCC. It is known that human placenta, brain, testis, ovary, spleen, and adrenal cortex express high levels of FACL4, whereas the gastrointestinal system, including liver, expresses a very low level.2) In a recent paper, 15) we reported that FACL4 is frequently upregulated in HCC compared to chronic hepatitis, as revealed by highthroughput screening combining RDA and microarray. We think that assessing the status of FACL4 in HCC is of importance to check whether the FACL4 pathway is also involved in liver carcinogenesis. Therefore, in this study, we investigated FACL4 upregulation in tumor cells of HCC and in cell lines derived from hepatoma. Materials and MethodsTumor samples and cell lines.