Reverse transcriptase-polymerase chain reaction (RT-PCR) for detection of occult malignancies in breast cancer patients is evolving as a useful diagnostic tool. However, no reliable molecular mRNA markers are available. We developed an RT-PCR plus Southern blot assay using p-hCG @-subunit of human chorionic gonadotropin) gene expression as a tumor marker for detection of breast malignancies metastatic to tumor-draining lymph nodes and blood. Breast carcinoma cell lines, primary breast malignancies and human placenta were used as positive controls for establishing the P-hCG RT-PCR assay. Peripheral blood leukocytes (PBL) from normal volunteer donors, normal breast tissue and lymph nodes from cancer-free patients were used as negative controls. P-hCG RT-PCR was used to assess tumor cell presence in PBL and tumor-draining axillary nodes from patients with AJCC stage I-IV breast cancer. The assay sensitivity and specificity were enhanced by restriction endonuclease digestion of an Sty I site of the RT-PCR cDNA product followed by Southern blot analysis. P-hCG mRNA was expressed in all breast cancer cell lines and 80% of primary breast cancers; it was not expressed in negative Neoplastic cells aberrantly express genes that are silent in their normal counterparts. These genes often encode for proteins that endow neoplastic cells selective advantages with respect to growth and other malignant cell properties, such as metastasis and invasion. Neoplastic cells produce autocrine growth factors or hormones that are not produced or are expressed at low levels by normal cells. Human chorionic gonadotropin (hCG) is a hormone produced by the trophoblast and is critical to sustaining pregnancy (Moyle and Campbell, 1993). hCG and its individual subunits are produced not only in gestational trophoblastic and germ-cell cancers but also in a wide range of non-gonadal tumors, such as gastrointestinal, lung and breast carcinomas (McManus et al., 1976; Madersbacher et al., 1994; Marcillac et al., 1992;Acevedo et al., 1995). We have demonstrated its usefulness as a molecular marker for melanoma (Doi et al., 1996). The pathophysiologic role for the expression of hCG in malignant neoplasms is currently speculative . hCG expression represents recapitulation of fetal cell properties by malignant cells.Immunohistochemical analysis using polyclonal or monoclonal antibodies (MAbs) has demonstrated hCG as a cancer marker (Krichevsky et al., 1995). However, these studies were not always specific, primarily due to the recognition of crossreactive epitopes or subunit chains of related gonadotropins (McManus et al., 1976; Krichevsky et al., 1995). About 7040% of adenocarcinomas arising in the mammary gland stain for the p subunit of hCG (P-hCG) by immunohistochemical techniques (Agnantis et al., 1992). hCG is a dimeric hormone composed of 2 non-covalently linked subunits designated a and P (Bo and Boime, 1992;Boorstein et al., 1982). It belongs to a glycoprotein hormone family, sharing many similarities with luteinizing hormone (LH), follicle stim...

Hepatocyte growth factor (HGF) is known to have a number of biological properties including promoting tumor progression of human carcinomas. Metastasis involves a number of events that are attributed to induction by paracrine factors such as HGF. Identification of natural inhibitors of these events would allow better control of tumor progression. Recently we demonstrated that interleukin 4 (IL-4) can regulate proliferation of various human carcinoma cell lines. In the present study, we used established human colon carcinoma cell lines and primary colon carcinoma cell cultures to determine if IL-4 could regulate HGF-induced cell proliferation and other events of tumor progression such as MMP (matrix metalloproteinases)-1, -2, and -9 production, cell migration and cell-matrix invasive activity. All colon carcinoma cell lines expressed HGF and IL-4 receptors. IL-4 significantly inhibited HGF-induced proliferation of one cell line. Cell-matrix invasion was significantly enhanced by HGF (0.1-10 ng/ml); IL-4 (1-10 U/ml) significantly inhibited HGF-induced invasion in a dose-dependent manner. IL-4 also inhibited HGF-induced cell-matrix invasion of metastatic colon carcinoma cells and HGF-induced cell migration. HGF enhanced MMP-1, -2, and -9 production by cell lines. This effect could be inhibited by IL-4. These findings indicate that IL-4 is a potent inhibitor of HGF-induced invasion and metastasis-related functions of human colon carcinoma cells.

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We investigated the pathogenesis of septic liver injury in rats caused by cecal ligation and puncture. In this model, numerous neutrophils accumulated in the liver in parallel with the development of liver dysfunction. The supernatants of hepatic macrophages isolated from these septic rats 24 hr after cecal ligation and puncture had enhanced chemotactic activities for human neutrophils. These results suggest that in sepsis, hepatic macrophages attract neutrophils to the liver. Human neutrophils preincubated in this macrophage supernatant had the following biological activities not seen in the sham-operated controls. (a) They became more adherent to cultured endothelial cells through up-regulation of adhesion molecules such as CD11b/CD18, (b) their chemiluminescence was markedly elevated. These functional changes of cecal ligation and puncture hepatic macrophages were the same as those in endotoxin-pretreated hepatic macrophages after isolation from normal rats. Therefore we suspect that hepatic macrophages are activated by portal vein endotoxin in sepsis. These activated hepatic macrophages secreted chemical mediators of inflammation, including leukotriene B4 and tumor necrosis factor. In conclusion, hepatic macrophages seem to interact closely with neutrophils and play an important role in the pathogenesis of septic liver injury.

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