Hiroki Odagiri

Upregulation of α-synuclein by lipopolysaccharide and interleukin-1 in human macrophages

α‐Synuclein was originally identified as the presynaptic nerve terminal protein. Recently, we reported that α‐synuclein is also expressed in cultured human astrocytes and that its levels are increased by stimulation with interleukin‐1β, suggesting that it may be involved in inflammatory processes. We therefore investigated the effect of inflammatory stimuli on α‐synuclein expression in human macrophages. α‐Synuclein mRNA and protein were detected in cultured human macrophages and levels of α‐synuclein protein were increased by stimulation with lipopolysaccharide and interleukin‐1β in a time‐ and concentration‐dependent manner. Immunofluorescent staining showed that α‐synuclein protein was expressed within the cytoplasm and nucleus. Furthermore, α‐synuclein immunoreactivity was present in alveolar macrophages from human lung tissues. These findings suggest that the function of α‐synuclein is not exclusive to the nervous system and that α‐synuclein may play a role in inflammatory processes and immune responses.

Cloned hst gene from normal human leukocyte DNA transforms NIH3T3 cells

The hst gene was originally identified as a transforming gene in DNAs from stomach cancers and a noncancerous portion of stomach mucosa by transfection assays using NIH3T3 cells (1,2). Subsequently, the hst gene obtained directly from leukocyte DNA of a leukemia patient was sequenced (3,4). Here, cosmid clones containing the hst gene were isolated directly from normal human leukocyte DNA and from T361-2nd-1 cells, a secondary transformant of NIH3T3 cells induced by transfection of DNA from a stomach cancer. All clones containing the hst gene from these different sources transformed NIH3T3 cells with similar efficiency. Restriction map of the hst gene from normal leukocyte DNA was identical with that from leukocyte DNA of a leukemia patient, while the hst gene from T361-2nd-1 cells was rearranged at the 168th nucleotide upstream of the TATA box.

Increased production of tumor necrosis factor and prostaglandin E2 by monocytes in cancer patients and its unique modulation by their plasma

SummaryWe investigated the role of monocytes in the production of tumor necrosis factor (TNF) and prostaglandin E2 (PGE2) in 77 cancer patients with malignancies of the digestive tract, using 30 normal individuals and 18 noncancer patients as controls. Monocytes were incubated with lipopolysaccharide for 20 h, and TNF production and PGE2 production were analyzed by bioassays. Elevated levels of TNF (>512 U/ml) and PGE2 (>8 ng/ml) production were demonstrated in many cancer patients when these factors were induced in the medium with 10% fetal bovine serum. The elevated level of TNF was seen to be restricted for the most part to patients with malignancies. Thus, 51 out of 59 cancer patients (86%), consisting of 44 primary cancer patients and 15 recurrent cancer patients, showed an increased level of TNF. In contrast, almost all of 18 postoperative cancer patients showed TNF levels comparable to those of normal individuals. Furthermore, 16 primary cancer patients were also demonstrated to have reduced levels of TNF production by monocytes after curative operation. When 10% cancer-patient plasma was added to the induction culture, TNF production by monocytes was drastically suppressed in the cancer patients. Interestingly, the same addition of plasma induced a prominent enhancement of PGE2 production in the cancer patients. The plasma of noncancer patients did not modulate production of these factors. No TNF activity was found in the plasma of cancer patients, but such plasma did contain an increased level of PGE2 (100–300 pg/ml). Although PGE2 (>2ng/ml) was able to suppress TNF production by monocytes, the addition of 10% plasma PGE2 was not enough to induce suppression. An unknown factor(s) in the plasma of cancer patients may uniquely modulate the elevated TNF and PGE2 production in these patients.

Bevacizumab in the treatment of five patients with breast cancer and brain metastases: Japan Breast Cancer Research Network-07 trial

Background Brain metastases from breast cancer occur in 20%–40% of patients, and the frequency has increased over time. New radiosensitizers and cytotoxic or cytostatic agents, and innovative techniques of drug delivery are still under investigation. Methods Five patients with brain metastases who did not respond to whole-brain radiotherapy and then received bevacizumab combined with paclitaxel were identified using our database of records between 2011 and 2012. The clinicopathological data and outcomes for these patients were then reviewed. Results The median time to disease progression was 86 days. Of five patients, two (40%) achieved a partial response, two had stable disease, and one had progressive disease. In addition, one patient with brain metastases had ptosis and diplopia due to metastases of the right extraocular muscles. However, not only the brain metastases, but also the ptosis and diplopia began to disappear after 1 month of treatment. The most common treatment-related adverse events (all grades) were hypertension (60%), neuropathy (40%), and proteinuria (20%). No grade 3 toxicity was seen. No intracranial hemorrhage was observed. Conclusion We present five patients with breast cancer and brain metastases, with benefits from systemic chemotherapy when combined with bevacizumab.

Anti-apoptotic effect of claudin-1 in tamoxifen-treated human breast cancer MCF-7 cells

BackgroundClaudin-1 is a membrane protein of tight junctions, and is associated with the development of various cancers. However, the significance of claudin-1 expression in cancer cells is not well understood. Here, we showed for the first time the anti-apoptotic effect of claudin-1 in human breast cancer MCF-7 cells.MethodsHuman breast cancer MCF-7 and T47 D cells were treated with or without tamoxifen, siRNA against claudin-1, or tamoxifen and claudin-1 siRNA. The samples were analyzed by RT-PCR, Western blotting or immunofluorescent staining.ResultsThe expression of claudin-1 was upregulated in tamoxifen-treated MCF-7 cells, whereas the expression of claudin-1 was not altered in tamoxifen-treated T47 D cells. Knockdown of claudin-1 by siRNA increased the amount of poly (ADP-ribose) polymerase (PARP) regardless of tamoxifen treatment in MCF-7 cells, but not T47 D cells. In the cell membranes of the MCF-7 cells, tamoxifen treatment increased the amount of claudin-1, but decreased the amount of β-catenin. Claudin-1 siRNA increased the amount of E-cadherin in the cytoplasm of the MCF-7 cells as well as the amount of β-catenin in their cell membranes.ConclusionThese results indicate that claudin-1 has anti-apoptotic effects, and is involved in the regulation of the expression and subcellular localization of β-catenin and E-cadherin in MCF-7, but not T47 D cells.

Aberrantly expressed recoverin is functionally associated with G-protein-coupled receptor kinases in cancer cell lines

Cancer-associated retinopathy (CAR) is an ocular manifestation of a paraneoplastic syndrome whereby immunological reactions toward recoverin (Rec), a retina-specific Ca(2+) binding protein, and its aberrant expression in tumor cells lead to the retinal degeneration. To elucidate functional roles of the aberrantly expression in cancer cells, we performed immunoprecipitation using anti-human Rec mAb. We observed co-precipitation of G-protein-coupled receptor kinases (GRKs) and caveolin-1 with Rec from cell lysates of 293 or SSTW cells. Immunocytochemistry revealed that immunoreactivities toward Rec within the cancer cells were almost identical to those toward GRKs and caveolin-1. The present data strongly suggest that aberrantly expressed Rec should be involved in the GRK-dependent cellular regulation in cancer cells.

Chemosensitivity of human pancreatic carcinoma cells is enhanced by IkBα super-repressor

Pancreatic cancer has an unfavorable prognosis; surgery and chemotherapy at present have only limited value. To improve the prognosis of pancreatic cancer, effective non‐surgical therapy is necessary. NF‐kB is reported to be related to resistance to apopto‐sis, but its role in Chemosensitivity remains controversial. We examined the effects on Chemosensitivity of inhibition by induction of the super‐repressor IkBα in pancreatic cancer cell lines, BxPC‐3, Capan‐1 and Panc‐1. IkBα protein was transduced by infection of adenovirus vector AxCAhlkBδN. Sensitivity to VP‐16 and doxorubicin was increased significantly by IkBα induction in all three pancreatic cell lines. To investigate molecular events during IkBα induction, we examined the changes in expression of drug‐resistance‐related genes by real‐time RT‐PCR and those in apoptosis‐related genes by cDNA microarray. There was no common change of gene expression before and after IkBα induction among the three pancreatic cancer cell lines, except for mdm2. Further examination of other genes is necessary for a better understanding of the molecular mechanisms of enhancement of Chemosensitivity through IkBα induction. However, we have confirmed that IkBα induction leads to an increase of Chemosensitivity of pancreatic cancer. Many problems remain before clinical application of this adenoviral system will be feasible, but our results may ultimately lead to an improved therapy of pancreatic cancer. (Cancer Sci 2003; 94: 467–472)

Efficacy of Vitamin E Treatment for Hand-Foot Syndrome in Patients Receiving Capecitabine

Capecitabine is a novel oral fluoropyrimidine that is converted within tumor cells to fluorouracil by thymidine phosphorylase [1]. Hand-foot syndrome (HFS) is the most frequent side effect of capecitabine and has been reported in up to 71% of patients receiving a starting dose of 1,250 mg/m2 twice daily [2, 3, 4, 5, 6, 7]. Grade 3 HFS was reported in up to 10–24% of patients. Treatment interruption and, if required, dose reduction usually ameliorate symptoms without compromising efficacy [8, 9]. Supportive treatments such as topical wound care, elevation, and cold compresses may help to relieve pain [10, 11]. Use of systemic corticosteroids, pyridoxine (vitamin B6), and cox-2 inhibitors have been used in patients developing HFS with cytotoxic agents including capecitabine and pegylated liposomal doxorubicin with varying success [11, 12, 13]. Kara et al. [13] from Turkey reported apparent benefit of vitamin E in managing HFS. Therefore we conducted this study to examine the efficacy of vitamin E in managing capecitabine-induced HFS. This retrospective, multicenter study was undertaken between 2005 and 2009 in HER2-negative patients with breast cancer treated with oral capecitabine 828 mg/m2 twice daily on days 1–21 every 4 weeks. Patients with symptoms of grade 2 HFS received oral vitamin E (Tocopherol Acetate, Eisai Pharmaceuticals, Tokyo, Japan) 100 mg/day without chemotherapy dose modification. Patient and treatment-related data, e.g. chemotherapy-related toxicities, dose of vitamin E, severity of symptoms, and tumor response to therapy, were recorded every 4 weeks. Patients underwent a complete der-matological examination at every visit. HFS including pain was graded using the National Cancer Institute Common Terminology Criteria for Adverse Events version 3.0 (NCI-CT-CAE v3.0). Wilcoxon signed rank test was used to examine patient demographics and treatment information. Median time to onset of grade 2 HFS was estimated. Severity of HFS was compared before and after vitamin E administration. We identified 32 patients developing grade 2 HFS during capecitabine therapy between January 2005 and February 2009, who subsequently received vitamin E with or without capecitabine treatment modification. The median time to first onset of HFS was 7.3 months (range 4.1–9.6). The initial starting dose of vitamin E for treatment of HFS was 100 mg/day, and the median dose of vitamin E was 200 mg (range 100–400 mg/day). Vitamin E application had a marked effect on dermatological complications within 7 days of initiation. The effect lasted throughout administration. Desquamation and pain reduced gradually (figs. ​(figs.1]1] and ​and2),2), and the comfort level of the patients improved. Fifteen of 32 patients (46.9%) with HFS experienced symptom improvement with vitamin E (100 mg/day) (p < 0.05; before vs. after 2 months vitamin E administration). Neurological symptoms improved. Thirteen patients still had pain, but this decreased after vitamin E dose escalation to 400 mg/day. The remaining 4 patients had considerable pain interfering with function after vitamin E 100 mg/day, but this reduced after vitamin E dose escalation to 400 mg/day and dose reduction of capecitabine as described previously [14]. Among all 32 patients included, the overall response rate to capecitabine was 37.5%, comprising 2 complete and 10 partial responses. Patients receiving capecitabine and vitamin E (100–400 mg) had longer time to progression than did patients receiving dose reduction of capecitabine (median 10.2 months vs. 6.1 months). Fig. 1 Hand-foot syndrome (HFS) began to disappear after 1 month of vitamin E treatment; 15 of 32 patients (47%) with HFS experienced symptom improvement with vitamin E (100 mg/day) (p < 0.05). Fig. 2 Clinical presentation of hand-foot syndrome. After vitamin E without dose reduction of capecitabine, the skin lesions had disappeared. In this retrospective study, 15 of 32 patients with HFS improved with vitamin E 100 mg/day, suggesting a beneficial effect of vitamin E therapy. Vitamin E is a widely used skin care product and functions as the major lipophilic antioxidant, preventing peroxidation of lipids and resulting in more stable cell membranes. The antioxidant membrane stabilizing effect of vitamin E also includes stabilization of the lysomal membrane, a function shared with glucocorticoids [13]. Systemic vitamin E and glucocorticoids inhibit the inflammatory response and collagen synthesis, thereby possibly impeding the

Japanese translation and linguistic validation of the US National Cancer Institute’s Patient-Reported Outcomes version of the Common Terminology Criteria for Adverse Events (PRO-CTCAE)

BackgroundThe US National Cancer Institute (NCI) has developed the Patient-Reported Outcomes version of the Common Terminology Criteria for Adverse Events (PRO-CTCAE) to capture patients’ self-reported symptomatic adverse events in cancer clinical trials. The aim of this study was to develop and linguistically validate a Japanese translation of PRO-CTCAE. Forward- and back-translations were produced, and an independent review was performed by the Japan Clinical Oncology Group (JCOG) Executive Committee and the US NCI. We then conducted cognitive interviews with 21 patients undergoing cancer treatment. Participants were asked to complete the PRO-CTCAE and were interviewed using semi-structured scripts and predetermined probes to investigate whether any items were difficult to understand or answer. The interviews were recorded and transcribed, and a thematic analysis was performed. The data were split into two categories: 1) remarks on the items and 2) remarks on the questionnaire in general.ResultsTwenty-one cancer patients undergoing chemotherapy or hormone therapy were interviewed at the University of Tokyo Hospital and the Kansai Medical University Hirakata Hospital during 2011 and 2012. Thirty-three PRO-CTCAE items were evaluated as “difficult to understand,” and 65 items were evaluated as “difficult to answer” by at least one respondent. However, on further investigation, only 24 remarks were categorized as “comprehension difficulties” or “clarity” issues. Most of these remarks concerned patients’ difficulties with rating their experience of individual symptomatic events.ConclusionsThe study provides preliminary evidence supporting the linguistic validity of the Japanese version of PRO-CTCAE. Further cognitive interviewing is warranted for PRO-CTCAE items relating to sexuality and anxiety and for response options on severity attribute items.

Efficacy of Hyperthermia in Combination with Radiation Therapy for Breast Cancer

Hyperthermia continues to show clinical benefits in randomized trials across a spectrum of malignancies, with generally well-tolerated side effects when administered as multimodality therapy. Hyperthermia contributes to direct tumor cell killing, and may also enhance the antitumor effects of chemotherapy and radiation therapy. Particularly, hyperthermia has been shown to be beneficial for treating superficial malignancies, such as chest wall recurrence of breast cancer. Although much evidence has been gained from clinical trials and studies of hyperthermia, regular use has not yet been established for breast cancer treatment. In this review article, we will highlight the benefits of therapeutic hyperthermia and the combined use of hyperthermia and radiation, and overview the results reported from clinical trials.