Proto-oncogene, Pim-3 with serine/threonine kinase activity, is aberrantly expressed in human colon cancer cells and can prevent Bad-mediated apoptosis

金沢大学がん研究所がん病態制御We previously observed that Pim-3 with serine/threonine kinase activity, was aberrantly expressed in malignant lesions of endoderm-derived organs, liver and pancreas. Because Pim-3 protein was not detected in normal colon mucosal tissues, we evaluated Pim-3 expression in malignant lesions of human colon, another endoderm-derived organ. Pim-3 was detected immunohistochemically in well-differentiated (43/68 cases) and moderately differentiated (23/41 cases) but not poorly differentiated colon adenocarcinomas (0/5 cases). Moreover, Pim-3 proteins were detected in adenoma (35/40 cases) and normal mucosa (26/111 cases), which are adjacent to adenocarcinoma. Pim-3 was constitutively expressed in SW480 cells and the transfection with Pim-3 short hairpin RNA promoted apoptosis. In the same cell line, a pro-apoptotic molecule, Bad, was phosphorylated at Ser112 and Ser 136 sites of phosphorylation that are representative of its inactive form. Ser112 but not Ser136 phosphorylation in this cell line was abrogated by Pim-3 knockdown. Furthermore, in human colon cancer tissues, Pim-3 co-localized with Bad in all cases (9/9) and with phospho-Ser112 Bad in most cases (6/9). These observations suggest that Pim-3 can inactivate Bad by phosphorylating its Ser112 in human colon cancer cells and thus may prevent apoptosis and promote progression of human colon cancer. © 2007 Japanese Cancer Association

High JC virus load in tongue carcinomas may be a risk factor for tongue tumorigenesis

The John Cunningham virus (JCV) asymptomatically infects a large proportion (~90%) of the population worldwide but may be activated in immunodeficient patients, resulting in progressive multifocal leukoencephalopathy. Recent reports demonstrated its oncogenic role in malignancies. In this paper, the presence of JCV-targeting T antigen was investigated in tongue carcinoma (TC, n = 39), dysplastic tongue epithelium (DTE, n = 15) and glossitis (n = 15) using real-time polymerase chain reaction (PCR) and in situ PCR and immunohistochemistry, and JCV copies were analyzed with the clinicopathological parameters of TCs. The results demonstrated that glossitis and DTEs had significantly lower copies of JCV (410.5 ± 44.3 and 658.3 ± 53.3 copies/μg DNA respectively) than TCs (981.5 ± 14.0, p  < 0.05). When they were divided into three groups with 0–200 copies/μg DNA (low), 201–1,000 (moderate) and more than 1001 (high), TCs showed 3 (7.6%) in the low group, 21 (53.8%) in the moderate group and 15 (38.4%) in the high group and glossitis showed 11 (73.3%) in the low group, 0 (0%) in the moderate group and 4 (26.6%) in the high group. The DTEs occupied an intermediate position between them (p < 0.001). In situ PCR demonstrated that the nuclei of TC and DTE cells are sporadically T-antigen positive but not in nasal turbinate epithelial cells. Immunohistochemistry for T-antigen protein revealed four positive cases only in TCs. The existence of JCV T-antigen DNA was not associated with the clinicopathological variables of TCs. In conclusion, the presence of JCV may be a risk factor of tongue carcinogenesis

Cytotoxic Activities, SAR and Anti-Invasion Effects of Butylphthalide Derivatives on Human Hepatocellular Carcinoma SMMC7721 Cells

A series of butylphthalide derivatives (BPDs) 1–8 were isolated from the extract of the dried rhizome of Ligusticum chuanxiong Hort. (Umbelliferae). The cytotoxic activities of BPDs 1–8 were evaluated using a panel of human cancer cell lines. In addition, the SAR analysis and potential anti-invasion activities were investigated. The sp2 carbons at C-7 and C-7a appeared to be essential for the cytotoxic activities of BPDs. BPDs 5 and 6 remarkably inhibited the migration and invasion of cancer cells. The anti-invasion activity of dimer 6 was demonstrated to be significantly higher than monomer 5

The oligonucleotide primer sequences and product size for PCR amplification.

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AIF-1-related molecules in lung squamous cell carcinoma.

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Expression of KAI1 and tenascin, and microvessel density are closely correlated with liver metastasis of gastrointestinal adenocarcinoma

AIM: To seek good markers to predict invasion and metastasis of gastrointestinal adenocarcinoma (GIA). METHODS: Expression of KAI1 and tenascin were examined on tissue microarrays containing gastric adenocarcinoma (n = 98), colorectal adenocarcinoma (n = 125), gastric adjacent non‐cancerous mucosa (n = 95) and colorectal adjacent non‐cancerous mucosa (n = 112) by immunostaining. Microvessel density (MVD) in GIA was labelled using anti‐CD34 antibody by immunostaining. Expression of KAI1 and tenascin, and MVD were compared with clinicopathological features of tumours, including PTEN (phosphatase and tensin homology deleted from human chromosome 10) and EMMPRIN (extracellular matrix metalloproteinase inducer) expression. RESULTS: KAI1 expression was higher in GIAs than in their adjacent non‐cancerous mucosa (p<0.05). KAI1 and tenascin expression showed a significantly negative association with liver metastasis of GIA (p<0.05), but not with depth of invasion, venous invasion or lymph node metastasis (p>0.05). A significantly negative relationship was observed between EMMPRIN and tenascin expression in GIA (p<0.05). MVD was positively correlated with depth of invasion, venous invasion, lymph node metastasis and liver metastasis of tumours (p<0.05), whereas it was negatively correlated with PTEN expression (p<0.05). CONCLUSIONS: Up‐regulated KAI1 expression may play an important part in malignant transformation of gastrointestinal epithelial cells. Reduced expression of KAI1 and tenascin might underlie the molecular basis of liver metastasis of GIA. Angiogenesis is a key event in the invasion and metastasis of GIA. These markers might be used to indicate liver metastasis of GIA in clinicopathological practice

Relationship between AIF-1, clinicopathological features,IL-6 and VEGF in NSCLC.

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SB203580 and ruxolitinib attenuated the activation of p38-MAPK signaling and JAK/STAT3 signaling by rAIF-1 in A549 cells.

(A) Left: Representative blots showing the expressions of p-p38, total p38, p-STAT3, total STAT3 and tubulin (internal control). Right: Averaged data showing the effects of 400 ng/mL rAIF-1 on the p-STAT3/ STAT3 ratio and p-p38/p38 ratio. Data are shown as mean ± SD. * P P P B) SB203580, an inhibitor of p38-MAPK signaling, attenuated the increase in the p-p38/p38 ratio in A549 cells induced by rAIF-1. Data are shown as mean ± SD. * P P C) Ruxolitinib, an inhibitor of JAK/STAT3 signaling, attenuated the elevation in the p-STAT3/STAT3 ratio in rAIF-1-treated A549 cells. Data are shown as mean ± SD. * P P P < 0.001.</p

Expression of AIF-1 in NSCLC.

(A) AIF-1 mRNA expression in NSCLC tissue obtained from TCGA database analyses. LUAD: Lung adenocarcinoma; LUSC: Lung SCC. Boxplots show median, interquartile range and range. **** P B) Kaplan-Meier analyses comparing overall survival (patients with stage II or stage II–IV lung SCC) and disease-specific survival (patients with stage II–IV or stage T3 lung SCC) between those with high AIF-1 expression (High) and those with low AIF-1 expression (Low). HR: Hazard ratio (with 95% confidence interval). (C) Pearson correlation analysis of the relation between AIF-1 expression and CD68 expression in patients with lung cancer, lung SCC (LUSC) and lung adenocarcinoma (LUAD). (D) Double immunofluorescence staining of NSCLC samples and adjacent tissues. AIF-1 (green) and CD68 (red) were co-localized (yellow) in lung macrophages around nests of tumor tissue. Nuclei are stained blue with DAPI. Scale bar: 100 μm.</p

Effects of rAIF-1 on A549 cell proliferation, migration and production of IL-6 and VEGF.

(A) Concentration-dependent effects of rAIF-1 on the mRNA expressions of IL-6 and VEGF. Data are shown as mean ± SD. *** P P B) Concentration-dependent effects of rAIF-1 on the protein expressions of IL-6 and VEGF. Data are shown as mean ± SD. * P P P C) Culture medium levels of IL-6 and VEGF secreted by A549 cells. Data are shown as mean ± SD. *** P P D) Viability of A549 cells treated with 100, 200 or 400 ng/mL rAIF-1 (CCK-8 assay). OD: Optical density. Data are shown as mean ± SD. **** P E) Migration of A549 cells treated with 400 ng/mL rAIF-1 (wound healing assay). Data are shown as mean ± SD. * P < 0.05.</p