IFN gamma-induced up-regulation of PD-ECGF/TP enhances the cytotoxicity of 5-fluorouracil and 5'-deoxy-5-fluorouridine in bladder cancer cells

BACKGROUND: PD-ECGF/TP is an essential enzyme in converting 5'DFUR and 5FU to their active metabolites and can be up-regulated by some cytokines. MATERIALS AND METHODS: PD-ECGF/TP mRNA and protein expressions were determined by RT-PCR and Western blot, respectively. The cytotoxicity of 5FU, 5'DFUR or MMC against RT-4 and T24 cells was evaluated by MTS assay. The PD-ECGF/TP expressions in primary bladder cancers were also analyzed. RESULTS: Levels of PD-ECGF/TP mRNA and protein were concomitantly elevated in RT-4 and T24 cells after IFN gamma treatment. IFN gamma decreased the IC50 of 5FU and 5'DFUR in both cell lines, while it did not alter the IC50 of MMC, which is not a substrate of PD-ECGF/TP. PD-ECGF/TP expression correlated with tumor stage and grade in primary bladder cancers. CONCLUSION: IFN gamma enhances the cytotoxicity of 5FU and 5'DFUR against human bladder cancer cells through induction of PD-ECGF/TP. The results imply that an IFN gamma/5FU or IFN gamma/5'DFUR combination therapy may be applicable to clinical bladder cancers.     

A case of recurrent advanced gastric cancer suggesting the efficacy of TS-1 and CDDP combination chemotherapy

The patient, a 53-year-old male, underwent radical surgery for advanced gastric cancer (stage IV). On the second day after surgery, adjuvant chemotherapy consisting of 250 mg/day 5-FU (i.v.) for 14 days, followed by 450 mg/day of UFT-E for about 12 months, was initiated. About 21 months after surgery (7 months after cessation of medication), the CA19-9 level had risen (136 U/ml). After 26 months, the patient experienced a backache and his CEA and CA19-9 levels had risen 11.7 ng/ml and 869 U/ml, respectively. The results from an imaging examination were suggestive of multiple bone metastases and para-aortic lymphatic metastasis. Chemotherapy was resumed with only TS-1 (100 mg/day). Because the tumor markers (TM) continued to rise, he was hospitalized and the medication was combined with daily administration of 10 mg of CDDP (TS-1 + CDDP protocol). When the total dose of CDDP reached 160 mg, there was a dramatic drop in the TM (surrogate marker) level. The patient was discharged and medication of TS-1 and 10 mg/day of CDDP twice a week was continued on an outpatient basis. Five months after the initial administration of FP, the CEA and CA19-9 returned to normal levels (4.3 ng/ml and 33 U/ml, respectively). Metastases to the para-aortic lymph nodes had disappeared and the sites of bone metastases were reduced in size. The patient was able to resume his full social activities. Since that time, a second-line therapy has been added. Currently (about two years after the recurrence), he is still undergoing therapy with TS-1 + CDDP.     

Serum cytokine level during continuous venovenous hemofiltration in toxic shock-like syndrome due to group G beta Streptococcus bacteremia in a patient with idiopathic thrombocytopenic purpura

We report a case of toxic shock-like syndrome due to a rare infection of group G Streptococcus bacteremia in a patient with idiopathic thrombocytopenic purpura and its successful treatment with continuous venovenous hemofiltration (CVVH). As the result of sepsis treatment with CVVH, in addition to administration of vasopressors and antibiotics, serum levels of interleukin-1beta, interleukin-10 and tumor necrosis factor-a fell and shock was controlled.     

Benign schwannoma of the breast: Report of a case

We report herein the case of a 62-year-old woman who presented to our hospital for investigation of occasional pain in her left breast. Although there was no mass palpable in her left breast, mammography and ultrasonography revealed a round tumor in the upper outer quadrant of the right breast. Although the mammography findings indicated that the tumor was benign, the possibility of a malignant neoplasm could not be ruled out by the ultrasonographic images. A final diagnosis of schwannoma was established by histopathological examination of the excised mass.     

A case report of hepatocellular carcinoma (Vp4)--an attempt to reduce residual tumor thrombus using combination therapy (hepatic arterial infusion, hepatic arterial embolization and radiation)

A 57-year-old man was found to have elevated levels of HCC markers during an observation of chronic hepatitis C. Diffused hepatoma was involved in the posterior lobe, and tumor thrombus extended into the main portal vein (Vp4). Posterior segmentectomy and tumor thrombectomy were performed. But, CT scan 45 days after the operation showed an enhancement at the residual tumor thrombus in the posterior branch. The patient received a hepatic arterial infusion of 5-FU, followed by hepatic arterial embolization. Then, we chose radiation therapy to the tumor thrombus. The most recent CT showed no enhancement at the reduced tumor thrombus. There have been almost no reports of treatment for residual portal thrombus. Careful observations are necessary in such patients.     

Safe and efficient transgene expression in rat hepatocytes induced by adenoviral administration into the biliary tract

We examined whether retrograde intrabiliary adenoviral administration could induce safe and efficient transgene expression in hepatocytes. We administered recombinant adenovirus carrying a reporter lacZ gene retrogradely into the common bile duct of rats and evaluated the transduction efficiency of the lacZ gene in the liver histochemically by X-gal staining, and also quantitatively by a chemiluminescent reporter gene assay. Retrograde administration of adenovirus into the common bile duct was shown to successfully induce transgene expression in the liver. Although transgene expression induced by intrabiliary adenoviral administration was observed predominantly at periportal areas, a considerable number of cells expressing the transgene were detectable even in lobular and centrilobular areas. Furthermore, histochemical analysis revealed that intrabiliary adenoviral administration resulted in gene transfer into hepatocytes, but not into biliary epithelial cells. Transgene expression in the liver was transient, and pathological and biochemical analyses revealed that hepatic damage caused by intrabiliary adenoviral administration was not substantial. The results demonstrated in the present study suggest that retrograde administration of adenovirus into the common bile duct can induce safe and efficient transgene expression in hepatocytes without causing considerable adverse effects, supporting the feasibility of adenovirus-mediated gene transfer into hepatocytes in clinical settings by means of endoscopic retrograde cholangiography.     

Adenovirus-mediated gene transfer into rat livers: comparative study of retrograde intrabiliary and antegrade intraportal administration

To examine the feasibility of liver-directed in vivo gene therapy, we administered recombinant adenoviruses carrying a reporter lacZ gene retrogradely into the common bile duct of rats, as well as antegradely into the portal vein. Transduction efficiency of the lacZ gene in the liver was estimated not only histochemically by X-gal staining, but also quantitatively by a chemiluminescent reporter gene assay. Retrograde infusion of adenoviruses into the common bile duct was shown to successfully induce transgene expression in the liver. Transduction efficiency induced by intrabiliary adenoviral administration was not significantly different from that induced by intraportal adenoviral administration. Although transgene expression induced not only by intraportal, but also by intrabiliary adenoviral administration was observed predominantly at periportal areas, a considerable number of cells expressing the transgene were detectable even in lobular and centrilobular areas. Mild infiltration of inflammatory cells into the liver and mild hyperplastic changes of hepatocytes were observed after intrabiliary and intraportal adenoviral administration. However, hepatic damage estimated pathologically was not substantial. Furthermore, although intrabiliary and intraportal adenoviral administration resulted in very mild elevation of liver-related serum biochemical parameters, apparent complications were not observed in any rats. Our results demonstrated in the present study suggest that retrograde administration of adenoviruses into the common bile duct can induce efficient transgene expression in the liver without causing severe adverse effects, supporting the feasibility of adenovirus-mediated gene transfer into the liver in clinical settings by means of endoscopic retrograde cholangiography.     

Simultaneous real-time detection of initiator- and effector-caspase activation by double fluorescence resonance energy transfer analysis

Fluorescence resonance energy transfer (FRET) with green fluorescent protein (GFP) variants has become widely used for biochemical research. In order to expand the choice of fluorescent range in FRET analysis, we designed various color versions of the FRET-based probes for caspase activity, in which the substrate sequence of the caspase was sandwiched by donor and acceptor fluorescent proteins, and studied the potential of these color versions as fluorescent indicators. Six color versions were constructed by a combination of cyan fluorescent protein (CFP), GFP, yellow fluorescent protein (YFP), and DsRed. Real-time monitoring in single cells revealed that all probes could detect caspase activation during tumor necrosis factor (TNF)-alpha-induced cell death as a fluorescent change. GFP-DsRed and YFP-DsRed were as sensitive as CFP-YFP, and CFP-DsRed also showed a large fluorescent change. By using two probes, CFP-DsRed and YFP-DsRed, we carried out simultaneous multi-FRET analysis and revealed that the initiator- and effector-caspases were activated almost simultaneously in TNF-alpha-induced cell death. These findings may give experimental bases for the development of novel techniques to analyze multi-events simultaneously in single cells by using FRET probes in combination.