Combination of laser microdissection, 2D-DIGE and MALDI-TOF MS to identify protein biomarkers to predict colorectal cancer spread.

Biomarkers are urgently required to support current histological staging to provide additional accuracy in stratifying colorectal cancer (CRC) patients according to risk of spread to properly assign adjuvant chemotherapy after surgery. Chemotherapy is given to patients with stage III to reduce the risk of recurrence but is controversial in stage II patients. Up to 25% of stage II patients will relapse within 5 years after tumor removal and when this occurs cure is seldom possible. The aim of this study was to identify protein biomarkers to stratify risk of spread of CRC patients. Laser micro-dissection was used to isolate cancer cells from primary colorectal tumors of stage II patients which did or did not metastasize within 5 years after surgical resection. Protein expression differences between two groups of tumors were profiled by 2D-DIGE with saturation CyDye labeling and identified using MALDI-TOF mass spectrometry. Evaluation of protein candidates was conducted using tissue micro array (TMA) immunohistochemistry on 125 colorectal tumor tissue samples of different stages. A total of 55 differentially expressed proteins were identified. Ten protein biomarkers were chosen based on p value and ratio between non metastasized and metastazised groups and evaluated on 125 tissues using TMA immunohistochemistry. Expression of HLAB, protein 14-3-3ß, LTBP3, ADAMTS2, JAG2 and NME2 on tumour cells was significantly associated with clinical parameters related to tumour progression, invasion and metastasis. Kaplan-Meier survival curve showed strong expression of six proteins was associated with good CRC specific survival. Expression of HLAB, ADAMTS2, LTBP3, JAG2 and NME2 on tumour cells, was associated with tumour progression and invasion, metastasis and CRC specific survival may serve as potential biomarkers to stratify CRC patients into low and high risk of tumour metastasis. Combined methods of laser microdissection, 2D DIGE with saturation labelling and MALDI-TOF MS proved to be resourceful techniques capable of identifying protein biomarkers to predict risk of spread of CRC to liver.

The New Zealand PIPER Project: colorectal cancer survival according to rurality, ethnicity and socioeconomic deprivation-results from a retrospective cohort study.

AIM: To investigate differences in survival after diagnosis with colorectal cancer (CRC) by rurality, ethnicity and deprivation. METHODS: In this retrospective cohort study, clinical records and National Collections data were merged for all patients diagnosed with CRC in New Zealand in 2007-2008. Prioritised ethnicity was classified using New Zealand Cancer Registry data; meshblock of residence at diagnosis was used to determine rurality and socioeconomic deprivation. RESULTS: Of the 4,950 patients included, 1,938 had died of CRC by May 2014. The five-year risks of death from CRC were: Maori 47%; Pacific 59%; non-Maori-non-Pacific (nMnP) 38%. After adjustment for demographic characteristics, comorbidity and disease stage at diagnosis, compared to nMnP the relative risk (RR) for Maori was 1.1 (95%CI: 0.8-1.3) and for Pacific 1.8 (95% CI: 1.4-2.5). We found no differences in risk of death from CRC by rurality, but some differences by deprivation. CONCLUSIONS: Disparity in outcome following diagnosis with CRC exists in New Zealand. Much of this disparity can be explained by stage of disease at diagnosis for Maori, but for Pacific peoples and those in deprived areas other factors may influence outcome. Further analyses of the PIPER data will explore the impact of any differences in management.

Excision of the seminal vesicles for locally advanced and recurrent rectal and sigmoid cancer.

BACKGROUND: This study aims to define the clinical and oncological outcome of 'en-bloc' excision of the seminal vesicles for locally advanced and recurrent tumours of the sigmoid and rectum. METHODS: Eight patients were identified from a prospective colorectal cancer database at a tertiary centre as having undergone excision of the seminal vesicles in continuity with a locally advanced or recurrent sigmoid or rectal adenocarcinoma. The presentation, operative details, histopathology, oncological outcome and morbidity of the procedure were assessed. RESULTS: Three patients were referred with recurrent tumours related to an anastomosis and five had a locally advanced sigmoid or rectal cancer. The need for resection of the seminal vesicles was determined from the preoperative pelvic magnetic resonance imaging scan or from an intraoperative finding of loss of the plane of dissection anterior to Denonvilliers' fascia. Restorative resection was achieved in all three patients where the primary tumour was located in the sigmoid or rectosigmoid, while all five patients with a rectal tumour had a permanent stoma. After a median follow-up of 43 months, seven patients are alive and disease-free and one patient has died of distant metastases. No patient has suffered a local recurrence. All five patients who were sexually active before surgery suffered from post-operative impotence. Two patients had temporary urinary retention with overflow. CONCLUSIONS: In carefully selected patients with locally advanced or recurrent rectal and sigmoid cancers that are attached to the seminal vesicles, en-bloc excision confers excellent local control but is associated with a high rate of sexual morbidity.

Methods of a national colorectal cancer cohort study: the PIPER Project.

AIM: Colorectal cancer is one of the most common cancers, and second-leading cause of cancer-related death, in New Zealand. The PIPER (Presentations, Investigations, Pathways, Evaluation, Rx [treatment]) project was undertaken to compare presentation, investigations, management and outcomes by rurality, ethnicity and deprivation. This paper reports the methods of the project, a comparison of PIPER patient diagnoses to the New Zealand Cancer Registry (NZCR) data, and the characteristics of the PIPER cohort. METHOD: National, retrospective cohort review of secondary care medical records (public and private) of all cases of ICD-10-AM C18-C20 on the NZCR in the calendar years 2007 and 2008 (main cohort) and an extended sample of Maori and Pacific cases, and non-Maori non-Pacific controls in 2006 and 2009 (extended cohort). RESULTS: Of the 6,387 patients identified from the NZCR 5,610 (88%) were eligible for PIPER. Reasons for exclusion were non-adenocarcinoma histology (3%) and non-colorectal primary (2%). Data were collected on 3,695 patients with colon cancer, 1,385 with rectal cancer and 466 with cancer of the recto sigmoid junction. CONCLUSIONS: The PIPER Project has generated comprehensive population level data detailing the diagnosis and management of colorectal adenocarcinoma in New Zealand. This will be used to assess the care provided to patients, and the impact of variations in care occurring between patient groups.

Treating pseudomyxoma peritonei without heated intraperitoneal chemotherapy--a first look in New Zealand.

BACKGROUND: Pseudomyxoma peritonei is a condition characterised by dissemination of mucin-producing neoplastic cells throughout the peritoneal cavity. There are two pathological subsets, disseminated peritoneal adenomucinosis and peritoneal mucinosis carcinomatosis. Once a lethal disease, cytoreductive surgery combined with heated intraperitoneal chemotherapy (HIPEC) is challenging debulking as the standard of care. OBJECTIVE: We present the first case series detailing the postoperative morbidity, mortality and survival outcomes of patients treated for pseudomyxoma peritonei by cytoreductive surgery without heated intraperitoneal chemotherapy by a single surgeon. DESIGN: Wellington Hospital clinical databases were retrospectively searched. Inclusion criteria were a diagnosis of pseudomyxoma peritonei with a major cytoreductive operation with the intention of complete cytoreductive clearance. Exclusion criteria were palliative debulking operations and patient records not available for analysis. RESULTS: 25 patients underwent cytoreductive surgery between June 1999 and July 2011. Mean follow-up was 43.5 months (1.5-138). Histological classification was DPAM for 13/25 and PMCA for 12/25. Complete cytoreduction (CC-0 and CC-1) was achieved in 21/25 patients. There was no 30 day mortality following primary cytoreduction. Six patients underwent subsequent debulking/cytoreductive surgery; one patient died following repeat surgery. Clavien-Dindo grade 3 or 4 complications occurred in 7/25 patients. Combined 5-year survival was 64%, 92% for DPAM and 33% for PMCA. CONCLUSION: Cytoreductive surgery alone may result in comparable survival outcomes to those achieved with combined surgery and HIPEC in selected patients, especially for patients with DPAM.

Overexpression of aminoacylase 1 is associated with colorectal cancer progression.

Aminoacylase 1 (ACY1) is a cytosolic enzyme responsible for amino acid deacylation during intracellular protein degradation. ACY1 has been implicated in a number of human tumor types. However, the exact role of ACY1 in tumor development remains elusive because it was found to be lost in small cell lung cancer and renal cell carcinoma but overexpressed in colorectal cancer (CRC). The present study aims to further clarify the relationship of ACY1 with CRC progression. Immunohistochemical staining was performed in tissue microarrays composed of 120 cases of CRC using a monoclonal anti-ACY1 antibody. Immunoreactivity was analyzed in association with patients' clinicopathologic parameters and survival time. The role of ACY1 in cell proliferation and apoptosis was assessed by silencing its expression in HCT116 cells using a small interfering RNA. Strong expression of ACY1 was found to be significantly associated with more advanced TNM stage, lymph node metastasis, positive vascular invasion, and shorter cancer-specific survival. ACY1 knockdown significantly inhibited cell proliferation and induced apoptosis. We concluded that ACY1 expression in CRC varies with stage and appears to play a role in cell proliferation and apoptosis. Further evaluation of ACY1 as a clinically useful prognostic marker and a potential drug target for CRC would seem worthwhile.

A study into external rectal anatomy: improving patient selection for radiotherapy for rectal cancer.

PURPOSE: This study was designed to determine the distance from the anal verge to the anterior peritoneal reflection in vivo, thereby improving the selection of patients for preoperative radiotherapy. METHODS: Measurement of the distance from the anal verge to the anterior peritoneal reflection, confluence of the taenia, and the origin of the sigmoid mesentery in 50 patients in the lithotomy position. RESULTS: The mean distance from the anal verge to the anterior peritoneal reflection was 11.9 cm (men) and 10 cm (women). To the origin of the sigmoid mesentery, the measurements were 18.8 cm (men) and 19.1 cm (women) and to the confluence of the taenia coli, 20.3 cm (men) and 18.8 cm (women). CONCLUSIONS: The distance from the anal verge to the origin of the sigmoid mesentery was approximately 19 cm in both men and women. Below this level tumors have limited mobility and should be amenable to radiotherapy.

Sexual function after rectal excision.

BACKGROUND: Rectal excision is associated with a risk of autonomic nerve damage and associated sexual dysfunction (SD). The evolution of our understanding of the anatomy and physiology of sexual function together with continual refinement of surgery for both benign and malignant disease has led to a decrease in the incidence of SD after rectal surgery. A knowledge of the degree of risk of postoperative SD is important both for the patient and as a benchmark for audit of individual colorectal practice. METHODS: The available literature on the anatomy, physiology and surgical aspects of this topic has been researched through the Medline database. The more recently available data are reviewed in the context of the historical evolution of surgery for benign and malignant rectal disease. RESULTS AND CONCLUSIONS: In the best hands, permanent impotence occurs in less than 2% of patients following restorative proctocolectomy and at a similarly low rate after proctocolectomy and ileostomy. Isolated ejaculatory dysfunction is also numerically a minor problem post operation for benign disease. Patient age is the most important predictor of SD after surgery for rectal cancer. The incidence of permanent impotence remains high (>40%) after abdomino-perineal excision of the rectum (APE) but the continued decline in the use of this operation in favour of low anterior resection (LAR), which carries about half the risk of impotence compared to sphincter ablating surgery, is likely to have resulted in a fall in the absolute number of patients rendered impotent as a result of rectal cancer surgery. Anatomical dissection of the pelvis with preservation of the named autonomic fibres results in a low and predictable rate of sexual morbidity. Surgeons could profitably spend more time with their patients discussing the possible effects of surgery on sexual function. Further research is required to determine the effects of adjuvant therapy for rectal cancer on sexual function.

Pleomorphic Angiomyolipoma of Digestive Tract: A Heretofore Unrecognized Entity.

Two cases of pleomorphic angiomyolipoma (AML) composed of bizarre epithelioid smooth muscle cells located in the gastrointestinal tract are reported. One involved the appendix of a 6-year-old girl, and the second the cecum of a 22-year-old woman. In both instances the tumor cells were immunoreactive for HMB-45 and A103. To the best of our knowledge, this peculiar variant of an AML has not previously been recognized in this location. Int J Surg Pathol 8(1):67-72, 2000