Adverse prognosis of epigenetic inactivation in RUNX3 gene at 1p36 in human pancreatic cancer.

Alteration in transforming growth factor-beta signalling pathway is one of the main causes of pancreatic cancer. The human runt-related transcription factor 3 gene (RUNX3) is an important component of this pathway. RUNX3 locus 1p36 is commonly deleted in a variety of human cancers, including pancreatic cancer. Therefore, we examined genetic and epigenetic alterations of RUNX3 in human pancreatic cancer. Thirty-two patients with pancreatic cancer were investigated in this study. We examined the methylation status of RUNX3 promoter region, loss of heterozygosity (LOH) at 1p36, and conducted a mutation analysis. The results were compared with clinicopathological data. Promoter hypermethylation was detected in 20 (62.5%) of 32 pancreatic cancer tissues, confirmed by sequence of bisulphite-treated DNA. Loss of heterozygosity was detected in 11 (34.3%) of 32 pancreatic cancers. In comparison with clinicopathological data, hypermethylation showed a relation with a worse prognosis (P=0.0143). Hypermethylation and LOH appear to be common mechanisms for inactivation of RUNX3 in pancreatic cancer. Therefore, RUNX3 may be an important tumour suppressor gene related to pancreatic cancer.

Suitability of a US3-inactivated HSV mutant (L1BR1) as an oncolytic virus for pancreatic cancer therapy.

Recently, the use of oncolytic viruses against cancer has attracted considerable attention. We studied the potential of the US3 locus-deficient herpes simplex virus (HSV), L1BR1, for oncolytic virus therapy. Its high specificity and potency indicate that L1BR1 is a promising candidate as a new oncolytic virus against pancreatic cancer. Moreover, the virus exhibited the unique characteristic of increasing apoptosis when used in combination with anticancer drugs. We assessed the feasibility of using the US3 locus-deficient HSV named L1BR1 as a new replication-competent oncolytic virus for the treatment of pancreatic cancer. The US3 locus of HSV has been shown to be a key gene in producing a multifunctional protein kinase that inhibits apoptosis induced by viral infections, chemicals and ultraviolet (UV) light. L1BR1 has been reported to be more than 10 000-fold less virulent than the parental virus in mice. In this study, we examined the tumor specificity and oncolytic effect of this attenuated replication-competent virus, L1BR1, in pancreatic cancers derived from SW1990, Capan2 and Bxpc-3cells compared with the parent virus and other well-known oncolytic herpes viruses (R3616 and hrR3). We also studied the efficacy of L1BR1 for the induction of apoptosis as an attribute of this virus in combination with the anticancer drugs 5FU and cisplatin. The combined treatment of the pancreatic cancer cells with L1BR1 and these anticancer drugs enhanced apoptosis significantly. More importantly, L1BR1 showed the lowest replication capacity in normal human hepatocytes, but the highest tumor-reducing effect in vivo among the oncolytic herpes viruses tested. In addition, L1BR1 significantly increased the induction of apoptosis of cancer cells when treated in combination with anticancer drugs although the parental virus inhibited the induction of apoptosis. These results suggest that L1BR1 is promising as a new anticancer oncolytic virus.

Normal hepatic hemodynamics during early postoperative period in recipients with adult live donor liver transplantation.

To recognize "normal" hepatic hemodynamics after live donor liver transplantation (LDLT), we analyzed Doppler parameters on recipients with a right liver graft and donors after extended left hepatectomy. Theoretically these values should be the same. From April 2000 to October 2004, 20 LDLTs were performed using a right liver graft. The 10 recipients without postoperative complications and their donors were included in this study. Portal venous velocity (PVV; cm/s), hepatic arterial peak systolic velocity (cm/s), and hepatic venous peak velocity (HVPV; cm/s) were measured during the first 2 weeks. In donors PVV and HVPV after LDLT were significantly higher after than before left hepatectomy: 19.2 +/- 4.2 vs. 31.5 +/- 13.0 cm/s (P = .013) and 23.0 +/- 7.2 vs. 41.8 +/- 10.3 cm/s respectively (P = .010). However, there were mild degrees of increased PVV and HVPV. In recipients, a markedly increased PVV (106.3 +/- 45.2 cm/s on day 1) was significantly higher than that in donors on each postoperative day. The hepatic arterial resistive index in recipients was also significantly higher than that in donors on each postoperative day, for example, 0.72 +/- 0.11 vs 0.62 +/- 0.04 on day 1 (P = .0326). In conclusion, we have shown "abnormal" hepatic hemodynamics in even those recipients without complications during the early postoperative period after LDLT.

Correlation between copy number of mitochondrial DNA and clinico-pathologic parameters of hepatocellular carcinoma.

AIMS: In the current study, we investigated possible correlations of the mtDNA copy number in hepatocellular carcinoma (HCC) with the pathological findings and prognosis. METHODS: We studied 31 HCC specimens using quantitative real-time polymerase chain reaction analysis, and the correlation between the mtDNA copy number and the clinicopathologic parameters and mutations in the D-loop region of the mitochondrial genome. RESULTS: The mtDNA copy number was reduced in HCCs compared with the corresponding non-cancerous liver tissues (p=0.002), and significantly correlated with tumour size (p=0.014) and cirrhosis (p=0.048). Patients with a low mtDNA copy number tended to show shorter 5-year survival rates than patients with a high mtDNA copy number when assessed by Kaplan-Meier curves, but not a significant (overall survival rate, 63 vs 83%; p=0.19). The copy number of HCC with mtDNA D-loop mutation or deletion was lower, but not significantly so (p=0.656, p=0.590, respectively). CONCLUSIONS: Our results indicated that a reduced copy number of mtDNA is correlated with HCC and associated with malignant potential.

PIVKA-II during perioperative period in patients with hepato-biliary-pancreatic diseases.

BACKGROUND/AIMS: In this study, we examined the influence of clinical treatments in the perioperative period upon PIVKA-II (plasma levels of protein induced by vitamin K absence or antagonist-II) in patients with hepatocellular carcinoma and pancreatobiliary diseases. METHODOLOGY: During a perioperative period, plasma PIVKA-II levels were measured in 144 patients with various hepatobiliary and pancreatic diseases by two types of EIA kit, using the conventional monoclonal antibody MU-3 and the new monoclonal antibody 19B7; Thrombotests were given at the same time. RESULTS: PIVKA-II of hepatocellular carcinoma patients showed higher reactivity with MU-3 than with 19B7. On the other hand, PIVKA-II of pancreatobiliary malignancies showed higher reactivity to 19B7 than it did to MU-3. In hepatocellular carcinoma patients, there was no correlation between PIVKA-II and Thrombotest levels; however, a significant correlation was found in patients with obstructive jaundice due to pancreatobiliary cancer. PIVKA-II levels decreased gradually to normal range within about 2 weeks after the curative operation. PIVKA-II levels in the patients receiving antibiotics containing N-methyl-thiotetrazole without administration of vitamin K remained high as long as the antibiotics were administered, but decreased to normal range as soon as the administration was finished. CONCLUSIONS: PIVKA-II which is produced by hepatocellular carcinoma may be different from that produced by obstructive jaundice due to pancreatobiliary malignancies. The clinical treatments in perioperative period, such as the administration of antibiotics containing N-methyl-thiotetrazole or vitamin K, readily influence PIVKA-II levels. Therefore, abnormal PIVKA-II levels must be carefully interpreted in the diagnosis of hepatocellular carcinoma and pancreatobiliary malignancies.

Clinical significance of alpha-fetoprotein mRNA during perioperative period in HCC.

BACKGROUND/AIMS: The presence of alpha-fetoprotein (AFP)mRNA in the peripheral blood of patients with HCC suggested that carcinoma cells still existed. We investigated the influence of an alternation of human AFPmRNA perioperatively detected in peripheral blood in matters of recurrence, prognosis and operative performance. METHODOLOGY: Thirty-three patients with HCC were enrolled for this study. AFPmRNA in peripheral blood before the operation, and on 1, 3, 7, 14, 21 and 28 post-operative days was examined by means of the reverse-transcriptase polymerase chain reaction (RT-PCR). RESULTS: In 18 of 33 patients (54.5%) with HCC, AFPmRNA was detected in peripheral blood pre-operatively. Ten of 22 (45.4%) who were detected with AFPmRNA in peripheral blood during the perioperative period were diagnosed with intrahepatic recurrence or distant metastasis within 12 months post-operatively. In contrast, 10 (90.8%) of 11 patients who were not detected with AFPmRNA in peripheral blood anytime during hospitalization were diagnosed as free of intrahepatic recurrence or distant metastases within 9 months after the operation. CONCLUSIONS: These results suggested that an alternation of AFPmRNA from HCC patients' peripheral blood during the perioperative period by means of this technique may be a practical tool in the detection of early recurrence or distant metastases after surgery.

Surgical treatment of intraductal papillary-mucinous tumors of the pancreas.

BACKGROUND/AIMS: IPMT (Intraductal papillary-mucinous tumor of the pancreas) is increasingly recognized. The aim of this study was to investigate the appropriate surgical treatment for these tumors. METHODOLOGY: Between January 1981 and September 1998, 62 patients with IPMT underwent surgery. We retrospectively examined the clinicopathological features and surgical outcomes of the patients. RESULTS: The types of IPMT were as follows: hyperplasia (20); adenoma (31); and carcinoma, both invasive (5) and noninvasive (6). Lymph node metastasis was found in 36% of the carcinomas. The size of mural nodules was more than 3 mm in all adenoma or carcinoma cases, while the percentage of hyperplasia less than 3 mm was 75%. Intraoperative pancreatoscopy and annular array ultrasonography were very useful, because they detected 10 lesions that could not be found by preoperative examinations, such as computed tomography, endoscopic retrograde pancreatography, and endoscopic ultrasonography. All patients underwent surgical resection, including 10 pancreaticoduodenectomies (Whipple's procedure), 10 pylorus-preserving pancreaticoduodenectomies, 13 pancreatic head resections with segmental duodenectomies, 17 distal pancreatectomies, 9 segmental resections of the pancreas, 2 duodenum-preserving pancreatic head resections, and 1 total pancreatectomy. No operative or hospital death was observed. The postoperative survival rate at 5 years was 71.6% for carcinoma in IPMT. All of the cases with hyperplasia, adenoma and noninvasive carcinoma survived. Only two of the patients with invasive carcinoma died. CONCLUSIONS: IPMT had a favorable prognosis, as compared with pancreatic duct carcinoma. When selecting a surgical procedure for treating these tumors, it is important to confirm the tumor extent, as well as the diagnosis of invasion or noninvasion. In cases with invasion, radical resection is required. On the other hand, organ-function-preserving procedures should be selected for diseases without invasion.

Peritoneal washings cytology combined with immunocytochemical staining in pancreatic cancer.

BACKGROUND/AIMS: Peritoneal washings from patients with pancreatic cancer demonstrate malignant cells in 0-58% of patients. But the significance of their potential for implantation and growth after radical surgery has not been clarified. METHODOLOGY: Peritoneal washings were collected from 74 consecutive pancreatic cancer patients during a 5-year period and studied by conventional staining and immunocytochemical staining using anti-CEA and anti-CA 19-9. Fifty of the 74 patients were resectable and the others were nonresectable. RESULTS: Seven out of 8 patients with macroscopic peritoneal metastasis were positive in conventional staining, but all 8 patients were positive in immunocytochemical staining. Five of the 66 (8%) patients without macroscopic peritoneal metastasis were positive in conventional staining, but the positive rate increased to 14 of 66 (22%) in immunocytochemical staining. There was no statistically significant difference in post-operative cumulative survival rate between the 13 positive cytology patients and 37 negative cytology patients without macroscopic peritoneal metastasis. CONCLUSIONS: The positive rate of peritoneal washings cytology was increased in this study by the use of immunocytochemical staining in addition to conventional staining. No statistically significant difference in post-operative cumulative survival rate between positive cytology patients and negative cytology patients without macroscopic peritoneal metastasis was observed.

Oncolytic virus therapy--foreword.

We are very pleased and proud to be able to publish this special issue of Current Cancer Drug Targets devoted to oncolytic virus therapy covering basic and clinical research on adenovirus, vaccinia virus, herpes virus, and Newcastle disease virus. In these papers, we welcome the world's top authorities in the field who have generously contributed their latest review articles for exclusive publication in this special issue. Moreover, this issue also includes a range of opinion from government drug organizations. Here we simply wish to bring together the newest knowledge and experience in the field of cutting-edge oncolytic virus therapy for researchers and every kind of cancer therapist. The Foreword presents a historical perspective on the development of oncolytic virus together with the encouraging results of recent clinical trials (e.g., H101 has been tested in clinical trial of nearly 250 patients and approved for human use by the Chinese FDA, while PV701 has been tried in over 110 patients, as described in our special issue).

Hypermethylation of multiple genes as clonal markers in multicentric hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is highly malignant and prone to multicentric occurrence. Differentiation between a true relapse of HCC and a second primary tumour appearing is of clinical importance. At this point, no convenient method is available to determine the origin of these HCCs. Tissue samples were obtained from 19 patients and analysed for the promoter hypermethylation status of multiple tumour suppressor genes (p16, DAP-Kinase, MGMT, GSTP1, APC, RIZ1, SFRP1, SFRP2, SFRP5, RUNX3, and SOCS1) using methylation-specific PCR (MSP). Methylation status was used to determine tumour clonality. In each of the 19 cases, at least one tumour was recognised as having an aberrantly methylated gene. The frequency of the methylation in tumour tissue was 57.1% in p16, 2.4% in DAP-kinase, 23.8% in GSTP1, 90.5% in APC, 45.2% in RIZ1, 64.3% in SFRP1, 59.5% in SFRP2, 28.6% in SFRP5, 47.6% in RUNX3, and 54.8% in SOCS1, while in MGMT, no aberrant methylation was detected. The methylation status of these genes was assessed using MSP as being either positive or negative, and was used to determine the tumour clonality. The clonality of every tumour could be decided even with lesions that could not be judged by clinical diagnosis or by another molecular method (mt DNA mutation). Determining the methylation status of multiple genes in multicentric HCC was useful as a clonal marker and provided useful information for characterising the tumour. From our findings, multicentric HCCs tend to occur more independently than metastatically from the original tumour. Expanded study should be pursued further for a better understanding of the molecular mechanism of hepatocarcinogenesis.

Lymph node metastasis in carcinoma of the body and tail of the pancreas.

BACKGROUND: There have been no precise reports concerning lymph node metastatic involvement in carcinoma of the body and tail of the pancreas. METHODS: Histopathological examination of lymph node involvement in 30 specimens obtained from patients who underwent pancreatic resection and wide dissection of lymph nodes, including para-aortic lymph nodes, for carcinoma of the body and tail of the pancreas was performed. RESULTS: Fourteen of 30 patients had lymph node involvement. The highest incidence of lymph node involvement was around the splenic artery (five of 30 patients), aorta (four of 30) and coeliac trunk (four of 30). No significant difference in survival rate between the lymph node-negative group and the lymph node-positive group was observed, but all patients in the positive group died within 2 years after surgery, and four patients with para-aortic lymph node involvement died from recurrence within 10 months after surgery. Survival rates were significantly worse in patients with histopathological extrapancreatic nerve plexus invasion, retropancreatic tissue invasion, tumour diameter more than 4 cm, histological portal system vein wall invasion and carcinoma invasion of the surgical margins. CONCLUSION: Although aggressive extended surgery including para-aortic node dissection has been performed, the postoperative survival rate is still low in patients with carcinoma of the body and tail of the pancreas. The high incidence of liver metastasis after surgery is a prime cause of the poor outcome, and effective therapy for postoperative liver recurrence requires evaluation.