For determining the mechanism of genetic control of <i>D-</i>penicillamine free-base-specific delayed-type hypersensitivity (DTH) in mice, footpad swelling response was employed. Studies with various congenic and recombinant inbred strains of mice revealed that <i>D</i>-penicillamine free-base-specific DTH was controlled by the I-A subregion. The footpad swelling response showed a highly antigen-specific pattern, and revealed that <i>D</i>-penicillamine free base was not cross-reactive with <i>D</i>-penicillamine disulfide or <i>L</i>-penicillamine, but with <i>D</i>-penicillamine HCl. Treatment of immune lymphoid cells with monoclonal antibodies plus complement revealed that the cells responsible for DTH transfer were Lyt-1<sup>+</sup>2<sup>––</sup>, L3T4<sup>+</sup> and la<sup>––</sup> T cells.
A mechanism responsible for the induction of NP-specific first order (inducer) suppressor cells (TS1) is described. TS1 cells are induced by i.v. administration of hapten-coupled splenic cells. Their activity is assessed by the adoptive transfer of NP-specific suppression during the afferent phase of the contact sensitivity response. NP-coupled firmly adherent, FcR+, I-A-bearing macrophages induce TS1. The antigen-presenting cells required for TS1 induction lack the Thy-1 and Lyt-1 markers, and are resistant to 500 R irradiation and to cyclophosphamide treatment. NP-coupled dendritic cells fail to induce TS1 activity. The induction of TS1 cells is genetically restricted by genes that map in the I-J region of the H-2 complex. The NP-coupled antigen-presenting cells must share at least one I-J allele with the TS1 donor for effective induction of TS1 activity. To minimize allogeneic effects in these studies, the activity of the TS1 population was assessed by adoptive transfer into syngeneic recipients. The present results are compared with the mechanisms required for the induction of second and third order suppressor cells.
Prostate cancer (PCa) is the most common malignant carcinoma that develops in men in Western countries. MicroRNA (miRNA) have the potential to be used as biomarkers and therapeutic targets for the treatment of various cancers. We found significantly higher expression of miR-30d in 3 PCa cell lines (PC3, DU145 and LNCaP) compared with 2 normal prostate cell lines (RWPE-1 and PrSc) using miRNA microarrays and qPCR. Clinicopathological study revealed that miR-30d expression levels were significantly higher in cancer tissue samples than in the paired normal controls (P = 0.03). Furthermore, the miR-30d-high group had shorter time to biochemical recurrence (P = 0.026). MiR-30d overexpressed PCa cells promoted proliferation and invasion in vitro. Inoculation of miR-30d depleted PCa cells dramatically reduced tumor volumes in vivo. Using reporter gene assay, we identified miR-30d as a downregulator of SOCS1 expression by directly binding to 3'-UTR of SOCS1. MiR-30d regulated the expression of phospho-STAT3, MMP-2 and MMP-9 through the downregulation of SOCS1. The levels of SOCS1 mRNA and protein were significantly down-regulated in prostate cancer tissues. Consistently, miR-30d expression was inversely correlated with SOCS1 expression (P = 0.03). The miR-30d-high/SOCS1-low group was associated with an increased risk of early biochemical recurrence (P = 0.0057). Taken together, miR-30d appears to be a novel independent prognostic marker of PCa progression that allows clinicians to identify patients who need more intensive treatments.
Although the incidence of renal neoplasms is not particularly high, their clinical presentation is often interesting. They tend to arise in association with familial neoplastic syndromes, i.e. renal cell carcinoma with von Hippel-Lindau disease. The cloning of the genes responsible for these hereditary pathological conditions revealed their involvement in sporadic renal cancers. Based on their molecular biological features, the classification of renal neoplasms was reassessed by the World Health Organization in 2004. In this chapter we discuss the relationships between the pathology and molecular biology of each histological type of renal neoplasm and the underlying molecular biological abnormalities.
Autopsy findings including immunohistochemical and ultrastructural study of intravascular bronchioloalveolar tumors (IVBAT) of the lung and liver which were incidentally found in a 68‐year‐old man were reported. The tumors presented as several, small nodular lesions in bilateral lungs and liver. Immunohistochemical study using the antibody against Factor VIII‐related antigen and electron microscopic study suggested endothelial nature of both pulmonary and hepatic lesions. The study by using anti‐estradiol antibody revealed the presence of estrogen in the cytoplasm of neoplastic cells, suggesting the possibility of the presence of estrogen receptors in the cells. Although the result is still preliminary, some role of estrogen in the development of IVBAT is suggested. The hepatic lesions have been thought to be metastases from the pulmonary IVBAT but the other possibilities such as primary hepatic or multicentric origin must be considered. Review of the previous reports of IVBAT disclosed several cases which were associated with liver involvement. ACTA PATHOL. JPN. 35 : 1453–1465, 1985.
MINT‐7014221: Cullin 2 (uniprotkb:Q13617) physically interacts (MI:0218) with Gag (uniprotkb:P05888), elongin C (uniprotkb:Q15370), elongin B (uniprotkb:Q15369), SOCS1 (uniprotkb:O15524) and RBX1 (uniprotkb:P62877) by pull‐down (MI:0096)
Aberrant crypt foci (ACF), consisting of morphologically irregular crypts, are thought to be precancerous lesions for colon cancers. For their molecular analysis, it is necessary to avoid contamination with adjacent normal crypts and stromal cells. Decreased hexosaminidase activity in ACF, which has been histochemically demonstrated, was used in the present study to classify isolated crypts in combination with morphological changes. The length, rim diameter, and width (average±SD, μm) of hexosaminidase‐positive (Hex+) crypts were 238.6±40.4, 89.5±22.9, and 57.6±14.0, respectively. For hexosaminidase‐negative (Hex‐) crypts, the values were 314.4±77.8, 140.3±45.7, and 97.3±34.7, the width being 1.69 tunes greater (P<0.0001). Crypts wider than 115 μm (approximately 2 tunes the average size of Hex+ crypts) were all from ACF, judging from hexosaminidase staining. To analyze transcription levels of Hex α and β subunits ( Hexa and Hexb , respectively), real‐tune relative quantitative reverse transcription‐polymerase chain reaction (RT‐PCR) analysis was performed using the LightCycler system. In aberrant crypts, both Hexa and Hexb were significantly down‐regulated to 0.266 (P<0.002) and 0.131 (P<0.001) units, respectively, compared with those in morphologically normal crypts, with β ‐actin as the internal standard. This decrease could be a molecular marker for precancerous enzyme‐altered ACF.
B8 The peptidyl-prolyl isomerase Pin1 regulates a subset of phosphorylated proteins by catalyzing the cis-trans isomerization of their specific phosphorylated Ser/Thr-Pro motifs. Pin1 is overexpressed in many types of human malignancies and may contribute to tumorigenesis by enhancing several oncogenic signaling pathways. Although Pin1 has been shown to be involved in cell transformation and in the maintenance of the malignant phenotype in prostate cancer, its specific substrates during these processes have not so far been determined. In our present study, we have utilized Pin1 as a molecular probe to capture cancer-specific phosphorylated proteins as diagnostic or prognostic markers. By the use of glutathione-S-transferase (GST) pull-downs and subsequent proteomic analyses using human prostate cancer and Dunning rat prostate cancer cells we have newly identified five prostate cancer-specific Pin1 binding proteins (PINBPs). We confirmed the up-regulation of these genes in prostate cancer cell lines and tissues, compared with non-cancerous cells and tissues. In addition, tissue micro-dissection based quantitative RT-PCR analysis of prostate cancer tissues further revealed that the expression of one of these Pin1-binding proteins (PINBP1) is closely associated with both a higher probability and a shorter period of tumor recurrence following surgery. These results indicate that Pin1-based proteomics analysis is a useful tool in the identification of cancer-specific phosphorylated proteins and that these factors are potential diagnostic and/or prognostic markers and anti-cancer targets.
The effect of low-intensity pulsed ultrasound (LIPUS) on cell growth was examined in three-dimensional-cultured chondrocytes with a collagen sponge. To elucidate the mechanisms underlying the mechanical activation of chondrocytes, intracellular signaling pathways through the Ras/mitogen-activated protein kinase (MAPK) and the integrin/phosphatidylinositol 3 kinase (PI3K)/Akt pathways as well as proteins involved in proliferation of chondrocytes were examined in LIPUS-treated chondrocytes.Articular cartilage tissue was obtained from the metatarso-phalangeal joints of freshly sacrificed pigs. Isolated chondrocytes mixed with collagen gel and culture medium composites were added to type-I collagen honeycomb sponges. Experimental cells were cultured with daily 20-minute exposures to LIPUS. The chondrocytes proliferated and a collagenous matrix was formed on the surface of the sponge. Cell counting, histological examinations, immunohistochemical analyses and western blotting analysis were performed.The rate of chondrocyte proliferation was slightly but significantly higher in the LIPUS group in comparison with the control group during the 2-week culture period. Western blot analysis showed intense staining of type-IX collagen, cyclin B1 and cyclin D1, phosphorylated focal adhesion kinase, and phosphorylated Akt in the LIPUS group in comparison with the control group. No differences were detected, however, in the MAPK, phosphorylated MAPK and type-II collagen levels.LIPUS promoted the proliferation of cultured chondrocytes and the production of type-IX collagen in a three-dimensional culture using a collagen sponge. In addition, the anabolic LIPUS signal transduction to the nucleus via the integrin/phosphatidylinositol 3-OH kinase/Akt pathway rather than the integrin/MAPK pathway was generally associated with cell proliferation.
