Membrane-type 1 matrix metalloproteinase (MT1-MMP) is known as an activator of MMP-2. We reported that the expression of MT1-MMP is a powerful indicator of lymph node metastasis in gastric cancers. ...We now examined the possibility that MT1-MMP expression also is an indicator of lymph node metastasis in breast cancer. We examined the clinicopathologic significance of the expression of MT1-MMP mRNA in 53 breast cancer specimens by the semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) assay. The highest expression of MT1-MMP was found in those specimens showing lymph node metastasis and/or lymph vessel invasion. Therefore, an analysis of MT1-MMP can predict the presence of lymph node metastasis in breast cancer patients by RT-PCR assay in minute amounts of tissue. We believe this is an objective assay for determining the role of MT1-MMP for metastasis in patients with breast cancer.
Elongation factor 1 gamma (EF1 gamma) is known to be a subunit of EF1, one of the G proteins that mediate the transport of aminoacyl tRNA to 80S ribosomes during translation. As little is known ...regarding the expression of EF1 gamma in human oesophageal carcinoma, this study looked at its expression using a northern blot analysis. Thirty six cases of oesophageal carcinoma and 15 oesophageal carcinoma cell lines were studied. The EF1 gamma mRNA overexpression at a level of twofold or more was seen in five (14%) of 36 carcinomatous tissues compared with the normal counterparts. All five overexpressed cases showed severe lymph node metastases compared with the non-overexpressed cases, and the difference was significant (p = 0.028). The stage of the disease of these five cases was far advanced compared with the nonoverexpressed cases (p = 0.012). All 15 oesophageal carcinoma cells expressed EF1 gamma mRNA relatively lower than the gastric or pancreatic carcinoma cell lines, in which EF1 gamma was originally isolated. As the expression of EF1 gamma mRNA could be detected even in the biopsy specimens, its overexpression in tumour tissue may provide preoperative useful information for predicting the aggressiveness of tumours.
The MAGE, BAGE, and GAGE genes code for distinct antigens that are recognized by autologous cytolytic T lymphocytes. We investigated the expression of these genes in both cell lines and surgical ...samples of gastric carcinoma, using reverse transcription-PCR. Furthermore, the induction of these genes by 5-aza-2'-deoxycytidine (DAC), a demethylating agent, was also examined in several cell lines. Of 11 cell lines, BAGE, GAGE1-6, GAGE1-2, MAGE-1, and MAGE-3 were detected in 7 (64%), 4 (36%), 3 (27%), 8 (73%), and 8 (73%) cell lines, respectively. After the in vitro treatment of the negative cell lines with DAC, the expression of these genes became positive in 46 to 91% of these cell lines. No expression of these genes was seen in any of the 57 samples of normal gastric tissue. In contrast, the tumor tissue samples expressed BAGE, GAGE1-6, GAGE1-2, MAGE-1, and MAGE-3 in 13 (23%), 9 (16%), 6 (11%), 25 (44%), and 23 (40%) tissue samples, respectively. Thus, at least one of these genes was expressed in 35 (61%) of 57 carcinomas. An analysis of the relationship between clinicopathological factors and the expression of these genes revealed that either BAGE or one of these genes was more frequently expressed in histologically intestinal-type than in diffuse-type carcinomas. Our results suggest that, because of the higher expression of these genes and the possible induction of these genes by DAC, patients with gastric carcinoma may, therefore, be potential candidates for tumor-specific immunotherapy directed against these antigens.
To estimate gastric cancer patients' prognosis more comprehensively,we tried to develop a prognostic scoring system using a cDNA microarray.
RNA was extracted from tumor/normal paired samples of 43 ...patients with gastric cancer, and cDNA microarray hybridization was performed.
We selected 78 genes that were differentially expressed between aggressive and nonaggressive groups with respect to five conventional pathological factors. Next, we determined a coefficient for each gene. Thereafter a prognostic score was calculated by summing-up the value for each gene. It ranged from -47 to 201 with a median of 114. There were two peaks in its distribution. Ten of 11 patients who were alive with no evidence of recurrence >5 years after the operation showed a score of <100 points, whereas all 19 patients who died of disease showed >100 points. In 13 patients who were alive but the follow-up time was <5 years, 2 of the 3 patients with >100 points revealed recurrent disease during the follow-up.
These findings demonstrate that such a system with cDNA microarray can contribute to the comprehensive analysis of malignant behavior of the tumor and may provide accurate information on prognosis.
Because the urinary excretion of drugs is often decreased in renal diseases, dosage regimens are adjusted to avoid adverse drug reactions. The aim of present study was to clarify the alteration in ...the levels of renal drug transporters and their correlation with the urinary drug excretion in renal diseases patients.
We quantified the mRNA levels of human organic anion transporters (hOATs) by real-time polymerase chain reaction and examined the excretion of the anionic drug, cefazolin, in renal disease patients. Moreover, transport of cefazolin by hOAT1 and hOAT3 were examined using HEK293 transfectants.
Among four hOATs, the level of hOAT1 mRNA was significantly lower in the kidney of patients with renal diseases than in the normal controls. The elimination constant of cefazolin showed a significant correlation with the values of phenolsulfonphthalein test and mRNA levels of hOAT3. The uptake study using HEK293 transfectants revealed that cefazolin and phenolsulfonphthalein were transported by hOAT3.
These results suggest that hOAT3 plays an important role for anionic drug secretion in patients with renal diseases and that the expression levels of drug transporters may be related to the alteration of renal drug secretion.
Insulin-like growth factor 2 (IGF2) stimulates cell proliferation and development in normal human growth. In several human cancers, the IGF2 gene is overexpressed and is thus considered to be a ...growth factor for tumors mediated through both the paracrine and autocrine pathways. However, the significance of IGF2 mRNA expression in gastric cancer has yet to be clarified. We semi-quantitatively measured the expression of IGF2 mRNA in 57 Japanese cases of gastric cancer by means of the reverse transcription polymerase chain reaction and also analyzed the relation between the IGF2 expression status and other clinicopathologic factors. We also performed immunohistochemical staining for IGF2. In 41 of 57 cases (72%), the expression of IGF2 mRNA was greater in tumor tissue (T) than in normal tissue (N). The average tumor/normal (T/N) expression ratio of IGF2 mRNA corrected for that of control gene mRNA was 1.42, while ranging from 0.36 to 3.65. The T/N ratio of infiltrative-type cancers was greater than that of expanding-type cancers (p<0.05). The cases with lymphatic permeation showed a greater T/N ratio than those without lymphatic permeation in expanding-type cancers (p<0. 05). Immunohistochemical staining revealed IGF2 to be detected in cancer cells themselves, especially at the margin of the cancer tissue. The IGF2 gene may thus play an important role in lymph vessel permeation especially in expanding-type gastric cancers.
There are few DNA-based studies that detect cancer micrometastases in lymph nodes. We have assayed for the specific detection of carcinoembryonic antigen (CEA)-expressing carcinoma cells in the lymph ...nodes of patients with gastrointestinal or breast carcinomas. A CEA-specific nested reverse transcriptase (RT)-PCR assay was optimized using limiting dilutions of a CEA-positive cancer cell line mixed with normal lymphocytes. The expression of CEA mRNA was studied in 100 carcinoma tissues, 75 normal mucosal tissues, and 15 lymph nodes from patients with cholelithiasis. Each of 117 lymph nodes from 13 patients with carcinoma was divided into two pieces: one was used for histological examination and the other for RT-PCR, and the results were compared. The sensitivity ratio was one CEA-expressing cancer cell detected in 1 x 10(5) normal lymphocytes. All carcinoma tissues and normal mucosal tissues expressed CEA mRNA, while no amplification was detected in any control lymph nodes. Thirty of 117 lymph nodes were histologically involved by carcinoma cells, and all of these yielded the expected product by RT-PCR. Of the remaining 87 histologically negative nodes, CEA mRNA was detected in 47 lymph nodes by RT-PCR. The positive rate increased from 26% by histological examination to 66% by RT-PCR. The assay by CEA-specific nested RT-PCR is not only sensitive but widely applicable for the detection of cancer micrometastases in lymph nodes. This method may lead to an earlier diagnosis and treatment of patients with subclinical lymph node metastasis.
Background: The clinical significance of pyrimidine nucleoside phosphorylase (PyNPase) activity in breast carcinomas has never been determined. Materials and methods: In 41 cases of breast carcinoma, ...the enzyme activity of PyNPase was determined by the high performance liquid chromatography (HPLC) assay and its value was analyzed with clinicopathologic variables. The expression level of mRNA was examined by the semi-quantitative reverse tran-scriptase-polymerase chain reaction (RT-PCR) assay and compared with the enzyme activity. Results: The higher activity of PyNPase was significantly correlated not only with the presence of vascular permeation (P = 0.02) but of lymph node metastasis (P = 0.02). The mRNA expression correlated well with the enzyme activity (r = 0.74, P < 0.01). A multivariate analysis disclosed the PyNPase factor to be associated with lymph node metastasis. In addition, 17 (41%) showed positive staining only in the tumor stromal cells and 18 (44%) cases showed positive staining in both the tumor stromal cells and the carcinoma cells by immunohistochemical study. Conclusions: These findings suggest that PyNPase activity is a new marker predicting the malignant potential of breast carcinomas, especially with respect to lymph node metastasis, and that the RT-PCR assay is a more useful method than direct evaluation of PyNPase activity.
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