High prevalence of diffuse large B-cell lymphoma in occult hepatitis B virus-infected patients in the Tohoku district in Eastern Japan.

Occult hepatitis B virus (HBV) infection is a clinical challenge, but its relationship to clinicopathologic features and the risk of progression to malignant lymphoma (ML) are poorly defined. We estimated the prevalence of HBV infection of 1,358 patients with newly diagnosed ML. HBV infection was more prevalent in ML than in control patients. The occult HBV infection group had a higher median onset age, no liver or spleen involvement, and higher prevalence of diffuse large B-cell lymphoma than the other groups, indicating that occult HBV infection is a distinct clinicopathologic entity. J. Med. Virol. 88:2206-2210, 2016. © 2016 Wiley Periodicals, Inc.

Methylation of the KEAP1 gene promoter region in human colorectal cancer.

BACKGROUND: The Keap1-Nrf2 pathway has been reported to be impaired in several cancers. However, the status of Keap1-Nrf2 system in human colorectal cancer (CRC) has not been elucidated. METHODS: We used colorectal cancer (CRC) cell lines and surgical specimens to investigate the methylation status of the KEAP1 promoter region as well as expression of Nrf2 and its downstream antioxidative stress genes, NQO-1 and AKR1C1. RESULTS: DNA sequencing analysis indicated that all mutations detected were synonymous, with no amino acid substitutions. We showed by bisulfite genomic sequencing and methylation-specific PCR that eight of 10 CRC cell lines had hypermethylated CpG islands in the KEAP1 promoter region. HT29 cells with a hypermethylated KEAP1 promoter resulted in decreased mRNA and protein expression but unmethylated Colo320DM cells showed higher expression levels. In addition, treatment with the DNA methyltransferase inhibitor 5-Aza-dC combined with the histone deacetylase inhibitor trichostatin A (TSA) increased KEAP1 mRNA expression. These result suggested that methylation of the KEAP1 promoter regulates its mRNA level. Time course analysis with the Nrf2-antioxidant response element (ARE) pathway activator t-BHQ treatment showed a rapid response within 24 h. HT29 cells had higher basal expression levels of NQO-1 and AKR1C1 mRNA than Colo320DM cells. Aberrant promoter methylation of KEAP1 was detected in 53% of tumor tissues and 25% of normal mucosae from 40 surgical CRC specimens, indicating that cancerous tissue showed increased methylation of the KEAP1 promoter region, conferring a protective effect against cytotoxic anticancer drugs. CONCLUSION: Hypermethylation of the KEAP1 promoter region suppressed its mRNA expression and increased nuclear Nrf2 and downstream ARE gene expression in CRC cells and tissues.

[A case of gastric cancer treated with modified docetaxel, cisplatin and 5-fluorouracil(mDCF)with ingestion inability].

The standard regimen of S-1 and cisplatin is not adaptable for patients with gastric cancer with an ingestion inability. A 74- year-old man was revealed to have unresectable gastric cancer with severe pyloric stenosis(cT4, cN3, cH1, cP0, cStage IV). He was treated with systemic chemotherapy using modified docetaxel, cisplatin and 5-fluorouracil(mDCF). He had manageable neutropenia(grade 3 and 4)during his treatment. CT findings after 3 courses showed reduced primary tumor and metastatic lesions. A curative operation was performed based on the effective response with downstaging. Palliative surgery was considered before receiving chemotherapy. mDCF therapy is one of the recommended options for gastric cancer with an ingestion inability.

Mesenchymal stromal cells promote tumor growth through the enhancement of neovascularization.

Mesenchymal stromal cells (MSCs), also called mesenchymal stem cells, migrate and function as stromal cells in tumor tissues. The effects of MSCs on tumor growth are controversial. In this study, we showed that MSCs increase proliferation of tumor cells in vitro and promote tumor growth in vivo. We also further analyzed the mechanisms that underlie these effects. For use in in vitro and in vivo experiments, we established a bone marrow-derived mesenchymal stromal cell line from cells isolated in C57BL/6 mice. Effects of murine MSCs on tumor cell proliferation in vitro were analyzed in a coculture model with B16-LacZ cells. Both coculture with MSCs and treatment with MSC-conditioned media led to enhanced growth of B16-LacZ cells, although the magnitude of growth stimulation in cocultured cells was greater than that of cells treated with conditioned media. Co-injection of B16-LacZ cells and MSCs into syngeneic mice led to increased tumor size compared with injection of B16-LacZ cells alone. Identical experiments using Lewis lung carcinoma (LLC) cells instead of B16-LacZ cells yielded similar results. Consistent with a role for neovascularization in MSC-mediated tumor growth, tumor vessel area was greater in tumors resulting from co-injection of B16-LacZ cells or LLCs with MSCs than in tumors induced by injection of cancer cells alone. Co-injected MSCs directly supported the tumor vasculature by localizing close to vascular walls and by expressing an endothelial marker. Furthermore, secretion of leukemia inhibitory factor, macrophage colony-stimulating factor, macrophage inflammatory protein-2 and vascular endothelial growth factor was increased in cocultures of MSCs and B16-LacZ cells compared with B16-LacZ cells alone. Together, these results indicate that MSCs promote tumor growth both in vitro and in vivo and suggest that tumor promotion in vivo may be attributable in part to enhanced angiogenesis.

[Survey on the compliance of patients with continuous infusion of 5-fluorouracil via portable infusion pumps].

A portable infusion pump is essential to sustain the 46-hour continuous administration of 5-fluorouracil in the folinic acid, fluorouracil, and oxaliplatin (FOLFOX) and folinic acid, fluorouracil, and irinotecan (FOLFIRI) protocols in colorectal cancer chemotherapy. However,the accuracy of the 5-fluorouracil dose administered via the infusion pump and patient compliance varies because the infusion rate changes depending on the viscosity of the drug, temperature, etc. In addition, the termination of administration based on the patient's judgment may influence these factors. In the present study, the amount of 5-fluorouracil remaining in the infusion pump and the administration time were investigated. As a result, the median amount that was found to remain in the pump was 49 mg, which was 2.0% of the average dosage, and an median administration time delay of 70 min was obtained. A questionnaire survey revealed that a majority of the patients felt insecurity about in adequate administration and administration time delays. These results indicate that customizing capacity modulation in the infusion pump corresponding to the patient's usage or seasonal variability of air temperature, and patient education may be important to improve patient compliance.

[A case of gastric adenosquamous carcinoma successfully treated with second-line chemotherapy (CPT-11 and CDDP)].

Gastric adenosquamous carcinoma is known as an infrequent histological cancer with a poor prognosis. A 69-year-old man was revealed to have gastric squamous carcinoma on the gastric body remote from esophagus (cT4 cN3, cStage IV). A curative operation was impossible so he was treated with systemic chemotherapy using S-1+docetaxel. After 1 course, we changed to second-line chemotherapy combining CPT-11+CDDP because of heterochronic multiple hepatic metastases. PET-CT and CT findings after 5 courses of second-line therapy showed reduced primary tumor and metastatic lesions. The curative operation was performed based on the effective response with downstaging. The final histological diagnosis showed gastric adenosquamous carcinoma. The adjuvant chemotherapy of CPT-11 was continued without relapse for almost 2 years.

Intratracheal delivery of CX3CL1-expressing mesenchymal stem cells to multiple lung tumors.

The lung is one of the organs to which cancers from solid tumors frequently metastasize. Multiple tumors in the lung are usually treated by systemic chemotherapy because of the lack of efficient methods of targeting antitumor agents to the lung. Although intratracheal administration is an ideal route for targeting multiple lung tumors, antitumor agents are often harmful to the organ or induce inflammation. Mesenchymal stem cells (MSCs), nonhematopoietic stem cells capable of differentiating into various mesoderm-type cells, have a propensity to migrate to and proliferate in tumor tissues after systemic administration. We intratracheally injected MSCs expressing CX3CL1 (MSC/RGDFKN) into the lung of lung tumor-bearing mice with multiple metastases of C26 or Lewis lung carcinoma (LLC). Antitumor effects were evaluated by counting the number of lung metastases and survival. We demonstrated the tropism of mouse MSCs to lung tumor tissues after intratracheal administration of GFP-positive MSCs. Intratracheal injection of MSC/RGDFKN strongly inhibited growth of lung metastases of C26 or LLC, and thus prolonged survival. Intratracheal injection of MSC/RGDFKN did not induce an inflammatory reaction in the lung. These results suggest that MSCs expressing antitumor agents can be delivered intratracheally into multiple lung tumor tissues without causing inflammation.

[A case of unresectable advanced gastric cancer with Trousseau syndrome].

Cancer-related coagulopathy is known as Trousseau syndrome with a poor prognosis. A 55-year-old woman was revealed to have gastric cancer (cT3 cN3, cStage IV) with deep vein thrombosis and pulmonary embolism. A curative operation was impossible so she was treated with systemic chemotherapy using S-1 combined with an anticoagulant agent (warfarin). Some studies indicated that the interaction between S-1 and warfarin might cause strong effects of their mutual interaction. Although the dose of warfarin had to be decreased, this patient was able to take both drugs safely without this severe adverse effect until a shift to palliative therapy.

Chronic nicotine stimulation modulates the immune response of mucosal T cells to Th1-dominant pattern via nAChR by upregulation of Th1-specific transcriptional factor.

Inflammatory bowel disease (IBD) consists of Crohn's disease (CD) and ulcerative colitis (UC). The etiology has not been clarified yet, but immune disorder is thought to be involved in the pathogenic physiology. Recently, general consensus has been reached that CD and UC are distinct, especially in respect of the immune response. Interestingly, smoking has diverse effects on CD, Th1-type enteritis, and on UC, Th2-type. However, the mechanisms remain obscure. Therefore, we hypothesized that nicotine altered the distinct immune responses in each form of IBD to affect their pathophysiology. In this study, we first demonstrated by RT-PCR analysis that human lamina propria T (LPT) cells had nicotinic acetylcholine receptor (nAChR), and express alpha7 nAChR subunit universally. In addition, the expression of T-bet mRNA in human LPT cells was significantly upregulated after the culture with 10(-7)M and 10(-5)M nicotine for 9 days, while chronic nicotine stimulation showed negligible effect on the expression of GATA-3 mRNA by real-time PCR. The effect of nicotine was inhibited by mecamylamine (MEC). These results suggested that nicotine could modulate the immune balance to Th1-dominant via nAChR in the intestine, to improve Th2-type enteritis. This may provide the experimental evidence for the fact that nicotine has a beneficial influence on UC, and exacerbates CD. Furthermore, it is of great interest that nicotine acts oppositely on CD and UC by modulation of the mucosal immune balance via the neurotransmitter receptor.

p53 negatively regulates intestinal immunity by delaying mucosal T cell cycling.

To mount an effective immune response, T cells must divide in response to antigen contact. To maintain tolerance, mucosal lamina propria T cells (LPTs) may adapt their cycling to an antigen-rich gut stimulatory environment. Here, we compared the cell cycle kinetics of LPTs and peripheral blood T cells (PBTs) before and after CD3- and CD2-mediated activation. While CD3-activated naive (CD45RA(+)) and memory (CD45RO(+)) PBTs peaked in the S and G2/M phase at 2-3 days, CD3-activated LPTs peaked at 4-6 days. In contrast, CD2 activation induced modest PBT but vigorous LPT cycling. The doubling time of CD3-activated PBTs was 1 day, while that of CD3- or CD2-activated LPTs was 2 days. LPTs failed to upregulate cyclin-dependent kinase 4 and cyclin D3, but Rb phosphorylation and cyclin A and B1 upregulation were induced by CD2 engagement. The extents of clonal expansion in LPT and PBT were comparable, indicating that LPTs' slow replication delays but does not hinder cell division. CD2-activated LPTs displayed a striking upregulation of p53, whose blockade by antisense oligonucleotides accelerated their S phase transit time to that of CD3-activated PBTs. By slowing LPT cycling, p53 may act as a negative regulator of mucosal immunity, promoting immunological tolerance by preventing excessive T cell replication.

[A case of unresectable advanced gallbladder cancer treated by systemic chemotherapy combined with hepatic arterial infusion].

A 53-year-old woman was revealed to have gallbladder cancer with liver metastases (H 1). Since a curative operation is impossible in this case, we started systemic chemotherapy employing S-1 combined with hepatic arterial infusion using epirubicin hydrochloride and mitomycin C. After three months, the primary lesion was reduced in size. The patient has been receiving systemic chemotherapy using S-1 only as an outpatient for 16 months. Although there is not enough evidence to support standard treatment, systemic chemotherapy combined with hepatic arterial infusion would improve the survival time without deterioration of quality of life in patients with advanced gallbladder cancer. This combined therapy should be considered one of the promising strategies for advanced gallbladder cancer.

[A case of unresectable advanced pancreatic cancer treated by S-1 with concurrent radiotherapy].

Chemotherapy (5-FU) with concurrent radiotherapy is recommended as an effective treatment for locally unresectable pancreatic cancer. A phase I study of S-1 with concurrent radiotherapy demonstrated promising results in late years. A 70-year-old man was revealed to have metastatic pancreatic cancer (T 4 N 3 M 1 (PER), Stage IVb). Since a curative operation was impossible in this case, he was treated with systemic chemotherapy using S-1 combined with irinotecan hydrochloride (CPT-11) as first-line chemotherapy. Because the primary lesion was increased in size after two courses,he was then treated by radiotherapy combined with S-1 as second-line treatment. S-1 (80 mg/body/day) was orally administered (2 consecutive weeks, 1-week break), and concurrent radiotherapy was performed at a daily fraction of 1.8 Gy, 5 days/week, total amount 45 Gy. Although in the early period of chemoradiotherapy, transfusion for anemia and morphine hydrochloride for pain control were necessary, his symptoms gradually improved by the reduction of primary lesion. The patient has been receiving systemic chemotherapy as an outpatient for 12 months without deterioration of quality of life.

[A case of gastric cancer with severe lymph node metastasis effectively treated with docetaxel / S-1 as neoadjuvant chemotherapy].

A 75-year-old male patient had advanced gastric cancer with severe lymph node metastasis. He was treated with docetaxel 60 mg/body (day 1) and S-1 120 mg/body (2 weeks administration and 1 week rest) as neoadjuvant chemotherapy. After two courses of this neoadjuvant chemotherapy, the primary lesion and lymph node swelling were remarkably improved. The patient underwent total gastrectomy and D2 lymph node dissection. The histological effect was judged to be Grade 3, and no viable cancer cell was detected in the primary lesion and lymph node (pCR). Docetaxel and S-1 combination therapy were thought to be an effective method as neoadjuvant chemotherapy for advanced gastric cancer with severe lymph node metastasis.

Sequential irinotecan hydrochloride/S-1 for S-1-resistant inoperable gastric cancer: A feasibility study.

Irinotecan hydrochloride (CPT-11) is reported to be involved in the downregulation of thymidylate synthase (TS), a target molecule of 5-fluorouracil (5-FU) and oral fluoropyrimidine S-1. Therefore, we hypothesized that a preceding administration of CPT-11 against S-1-resistant tumors may recover sensitivity to S-1. To this end, we planned a S-1/CPT-11 sequential therapy as a feasibility study in S-1-refractory gastric cancer patients. In the first course, CPT-11 was administered intravenously at 150 mg/m(2) on days 1 and 15. Subsequently, S-1 was administered orally for 4 weeks from day 29 to 57, followed by a 2-week interval (sequential S-1/CPT-11). When the tumor showed a complete response (CR) or partial response (PR), the same dose of S-1 monotherapy was continued unless progressive disease (PD) was observed. When the response was stable disease (SD), S-1 was administered at the same dose for just 2 weeks (days 1-15), no drug was administered for the following 2 weeks (4-week cycle) and CPT-11 was administered intravenously at 100 mg/m(2) on days 1 and 15 (concurrent S-1/CPT-11) unless PD was observed. In the case of PD, the study was terminated. The primary endpoint was an antitumor effect and secondary endpoints were median survival time (MST), progression-free survival (PFS), time-to-treatment failure (TTF) and safety. The response rate (RR) following the first course was only 5.9% and the most positive RR was 11.8%. The MST, median TTF and PFS were 381, 69 and 71 days, respectively. Leukocytopenia was observed in more than half of the patients. Since the RR was lower than estimated in an interim analysis, the trial was terminated and the protocol was concluded to be unfeasible.

Comparison of long-term clinical outcomes of CHOP chemotherapy between Japanese patients with nodal peripheral T-cell lymphomas and those with diffuse large B-cell lymphoma in the study group of the Tohoku Hematology Forum.

To clarify the clinical outcome of peripheral T-cell lymphomas (PTCLs), we conducted a retrospective review comparing the outcomes of patients with PTCL (nodal peripheral T-cell lymphoma, unspecified, n=34 ; angioimmunoblastic T-cell lymphoma, n=12) to those with diffuse large B-cell lymphoma (DLBCL, n=48). All patients received CHOP-based chemotherapy without rituximab. PTCL patients presented at a more advanced clinical stage (91% vs. 65%, P<0.002) with a poorer performance status (26% vs. 17%, P<0.002) than DLBCL patients. The complete response rate among PTCL patients was significantly lower than among DLBCL patients (39% vs. 67%, P<0.008), as was the 3-year overall survival rate (26% vs. 50%, P=0.005), and Cox multivariate analysis revealed immunophenotype, performance status, and extranodal site involved to be significantly associated with shorter overall survival (P=0.045, P=0.007, and P=0.034, respectively). Our findings suggest that PTCL patients tend to have a poor prognosis associated with several initial risk factors. Moreover, the T-cell phenotype itself appears to have a significant impact on overall survival. Thus, standard CHOP chemotherapy may be inadequate for PTCLs, especially in patients with high-risk factors. The development of newly stratified therapies for the treatment of PTCLs would be highly desirable.