Suzanne C. Schiffman

Overall survival peri-hilar cholangiocarcinoma: R1 resection with curative intent compared to primary endoscopic therapy†

Patients with peri‐hilar cholangiocarcinoma who undergo R1 resection with curative intent will have an improved survival compared to patients who were not resected.

Factors associated with recurrence and survival following hepatectomy for large hepatocellular carcinoma: A multicenter analysis

Optimal management of large (>5 cm) hepatocellular carcinoma (HCC) remains controversial. We sought to determine the factors associated with recurrence and survival for patients with large HCC following hepatectomy.

The association of manganese superoxide dismutase expression in Barrett's esophageal progression with MnTBAP and curcumin oil therapy.

BACKGROUND The aim of this study was to investigate the relationship between reflux induced bile insult and MnSOD expression, as well as to examine therapies to preserve MnSOD expression. Additionally, we sought to examine the relationship between MnSOD protein expression and MnSOD enzymatic activity. METHODS MnSOD protein expression was determined by Western blot assay and enzymatic activity was determined by SOD assay. The enzymatic activity of the Het-1A and Bar-T cells were compared both before and after treatments. RESULTS MnSOD expression in Het-1A cells was decreased after bile salt exposure. The cells that received MnTBAP or curcumin oil pretreatment showed increased MnSOD expression compared with control untreated cells. The Bar-T cells showed an increase in MnSOD expression after treatment with bile salts. The cells that were pretreated with MnTBAP displayed a larger increase in MnSOD expression compared with the cells that were not pretreated prior to bile salt exposure. The MnSOD activity was significantly different between the untreated cell lines (P = 0.01) and after treatment with bile salt (P = 0.03). Additionally, Bar-T cells had significantly less MnSOD activity than Het-1A cells after each of the pretreatments. CONCLUSIONS We demonstrated preservation of MnSOD expression in Het-1A cells that were pretreated with antioxidants including MnTBAP, curcumin oil, and certain berry extracts. Additionally, we demonstrated that Bar-T cells have significantly less MnSOD activity than Het-1A cells. These finding have important implications for future studies regarding chemoprevention and the treatment of esophageal cancer.

The Resistance of Esophageal Adenocarcinoma to Bile Salt Insult is Associated with Manganese Superoxide Dismutase Expression

BACKGROUND Bile acids are implicated as etiologic agents in esophageal cancer. We sought to analyze the impact of bile acid exposure on esophageal epithelial cells, Barretts metaplastic cells (BE), esophageal adenocarcinoma cells (EAC), and esophageal squamous carcinoma cell (ESC). We sought to determine if cellular resistance is related to manganese superoxide dismutase expression. METHODS Cells were exposed to sodium choleate (CA), sodium deoxycholate (DCA), sodium glycocholate (GCA), sodium taurocholate (TCA), or a 1:1 mixture (MIX) of reagents at concentrations in the range 0.2-0.8 mM. Cell viability was evaluated by MTT assay. Manganese superoxide dismutase (MnSOD) expression was analyzed by Western blot. Statistical analysis was performed using SPSS ver. 17.0, SPSS Inc., Chicago, IL. RESULTS Bile salt exposure inhibited cell viability in esophageal squamous cells in time- and growth-dependent manner. There was a 50% decrease in cell viability from 4 to 24 h. BE, EAC, and ESC cell lines were more resistant to bile insult. In untreated cell lines, MnSOD expression was significantly decreased in EAC and ESC cell lines compared with esophageal squamous epithelial cells and BE cells (P=0.002). Exposure of ESC cells to bile salt increased MnSOD expression. CONCLUSION The confirmation of the role of reactive oxygen species (ROS) and bile acids in esophageal carcinogenesis has interesting implications for chemoprevention in patients with reflux esophagitis and Barretts esophagus. Further studies are necessary to assess the preventative role of antioxidant supplementation.

Effect of freeze-dried berries on the development of reflux-induced esophageal adenocarcinoma.

The incidence of esophageal adenocarcinoma in humans is increasing more rapidly than any other malignancy in the United States. Animal studies have demonstrated the efficacy of freeze-dried berry supplementation on carcinogen-induced esophageal squamous cell carcinoma in rats; however, no such studies have been done in esophagoduodenal anastomosis (EDA), an animal model for reflux-induced esophageal adenocarcinoma (EAC) development. Eight-week-old male Sprague-Dawley rats were randomized into 3 groups: EDA + control diet (EDA-CD; n = 10); EDA + 2.5% black raspberry diet (EDA-BRB; n = 11) and EDA + 2.5% blueberry diet (EDA-BB; n = 12). After 2 wk of feeding the respective diets, the rats underwent EDA surgery to induce gastroesophageal reflux and then continued the diet. Measurement of feed intake suggested that all EDA-operated animals had lower feed intake starting at 10 wk after surgery and this was significant close to termination at 24 wk. There were no significant differences in either reflux esophagitis (RE), intestinal metaplasia (IM) (70% in CD, 64% in BRB, and 66% in BB; P = 0.1) or EAC incidence (30% for CD, 34% for BRB, and 25% for BB; P = 0.2) with supplementation. Berry diets did not alter COX-2 levels, but BB diet significantly reduced MnSOD levels (1.23 ± 0.2) compared to control diet (2.05 ± 0.14; P < 0.05). We conclude that a dietary supplementation of freeze-dried BRB and BB at 2.5% (w/w) was not effective in the prevention of reflux-induced esophageal adenocarcinoma in this EDA animal model.

Is prior cholecystectomy associated with decreased survival in patients with resectable pancreatic adenocarcinoma following pancreaticoduodenectomy

BACKGROUND Patients with pancreatic cancer who present with biliary symptoms may undergo cholecystectomy and thus delay cancer diagnosis. We hypothesized that prior cholecystectomy leads to decreased overall survival in patients with pancreatic adenocarcinoma. METHODS Retrospective study of hepatobiliary database. RESULTS Three hundred sixty-five patients with a diagnosis of resectable periampullary pancreatic adenocarcinoma were identified. Eighty-seven patients underwent prior cholecystectomy. Median age (P = .48), body mass index (BMI) (P = .8), diabetes status (P = .06), American Society of Anesthesiologists (ASA) class (P = .22), stent placement (P = .13), operative time (P = .76), estimated blood loss (EBL) (P = .24), intraoperative transfusion (P = .91), portal vein resection (P = .25), LOS (P = .09) adjuvant therapy (P = .2), tumor size (P = .89), differentiation (P = .67), angiolymphatic invasion (P = .69), perineural invasion (P = 54), nodal metastasis (P = .43), complication rate (P = .75), and 30-day mortality (P = .58) were not statistically different between patients with previous cholecystectomy and those without. Median survival was 14 months for patients with a history of cholecystectomy and 16 months for those without (P = .25). Previous cholecystectomy was not a predictor of survival on Cox regression analysis. CONCLUSION There was no difference in overall survival in patients with pancreatic cancer with prior cholecystectomy versus those without.

Abstract 4077: Exposure to bile acids alters intracellular expression of MnSOD in Barrett's esophagus

Introduction: Oxidative stress secondary to bile acid exposure has been associated with metaplastic degeneration of normal esophageal mucosa into Barrett9s Esophagus cells and eventually Esophageal Adenocarcinoma. We have previously reported that the macromolecular response of BE cells to this stress is largely regulated by the expression of Manganese Superoxide Dismutase (MnSOD). As the mitochondrion plays a vital role in MnSOD activation, the purpose of this study was to determine the location and activity of MnSOD within the BE cell after exposure to oxidative stress. Methods: BE cells were cultured to 80-90% confluence in 75ml flasks. The cells were then exposed for four hours to 0.4mM concentrations of taurocholic acid (TCA) or a 0.4mM 1:1 mixture of bile salts including sodium cholate (CA), sodium deoxycholate (DCA), sodium glycocholate (GCA), and sodium taurocholate (TCA) to mimic natural biliary refluxate. Proteins were extracted and separated into mitochondrial, nuclear, and cytoplasmic fragments followed by western blot analysis. Enzymatic Activity of MnSOD was determined by silencing of other SOD molecules with potassium cyanate. Lethality testing of biliary exposure was performed with MTT assays. Results: Mitochondrial expression of MnSOD was significantly increased in the cells exposed to a mixture of bile acids vs. TCA and control. This was present on both qualitative and quantitative analyses of Western blots. Cytoplasmic expression of MnSOD was also present and conserved between treated and untreated cells. Control cells lines showed no significant change in baseline MnSOD expression. This expression was confirmed by increased MnSOD activity in the mitochondria of cells exposed to mixed bile. The lethality of mixed bile for Barrett9s cells was confirmed by MTT assay, with isolated bile salts and controls having a significantly higher fraction of living cells. Conclusion: This is the first study to evaluate microcellular MnSOD expression which is increased in BE cells in response to the oxidative stress of bile exposure, specifically in the mitochondrion. Mitochondrial MnSOD activity is increased within the BE cell after exposure to bile. Further investigation is required to determine the potential correlation between bile exposure and BE to adenocarcinoma progression. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4077. doi:10.1158/1538-7445.AM2011-4077

1041 Hepatectomy is Superior to Thermal Ablation for Patients With a Solitary Colorectal Liver Metastasis

Introduction Hepatic resection is the mainstay of treatment for solitary colorectal liver metastases (mCRC); however, some patients are not ideal candidates. The aim of this study was to compare outcomes for patients with solitary mCRC who underwent resection or ablation.

Abstract 2074: Association of MnSOD and microRNA-301a regulation with inhibition of pancreatic ductal adenocarcinoma

Proceedings: AACR 101st Annual Meeting 2010‐‐ Apr 17‐21, 2010; Washington, DC Background: Pancreatic cancer is the fourth leading cause of cancer-related death in the United States. Manganese superoxide dismutase (MnSOD) has been found to be low in human pancreatic ductal adenocarcinoma (PDAC). Overexpressing MnSOD in PDAC has decreased growth rate of cancer cells. microRNAs, short 20-25 nucleotide RNA molecules, can function as oncogenes or as tumour suppressors. It has been shown that microRNA-301a is specifically up-regulated in PDAC specimen and inhibition of microRNA-301a has a decreased growth rate of PDAC cells. Aim: This study is to determine the relationship between the level of MnSOD and microRNA-301a regulation regarding the growth inhibition of PDAC. Methods: we carried out two experiments. A total of 60 PDAC specimens, along with 20 normal pancreatic tissues were obtained for immunohistochemistry (IHC) and in situ hybridization (ISH) studies to evaluate MnSOD and microRNA-301a expression. All pathology findings for the specimens were scored by two pathologists who were both blinded to the subjects. We also established a xenograft PDAC mouse model to study the growth rate in vivo. Nude BALB/C mice, 6 weeks old, were injected subcutaneously with 5×106 human PDAC (PanC-1 cells transfected with micro-301a antisence and control). Tumor-bearing animals were maintained for 8 weeks. Tumor growth was followed and tumor volume (TV) was determined using the standard formula, TV = (L xW2)/2. Results: There were significant decreases of MnSOD expression in PDAC specimens (1.5±0.06) compared to normal controls (2.5±0.07; p < 0.05). microRNA-301a expression was significantly increased in PDAC specimens (0.85±0.046) compared to normal controls (0.1±0.05; p < 0.05). The animal study findings indicated that microRNA-301a antisence transfection can significantly decrease the growth rate of inoculated PDAC cells. The decrease the growth rate is associated with the increased MnSOD levels (Fig 1) and decreased NF-kappaB expression. Conclusions: 1) MnSOD levels are negatively associated microRNA-301a expression in PDAC tissues; 2) increased levels of MnSOD are associated with microRNA-301a down-regulation and inhibition of PDAC. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2074.