Correlation of Chromosomal Instability,Telomere Length and Telomere Maintenance in Microsatellite Stable Rectal Cancer: A Molecular Subclass of Rectal Cancer

Introduction

Colorectal cancer (CRC) tumor DNA is characterized by chromosomal damage termed chromosomal instability (CIN) and excessively shortened telomeres. Up to 80% of CRC is microsatellite stable (MSS) and is historically considered to be chromosomally unstable (CIN+). However, tumor phenotyping depicts some MSS CRC with little or no genetic changes, thus being chromosomally stable (CIN-). MSS CIN- tumors have not been assessed for telomere attrition.

Experimental Design

MSS rectal cancers from patients ≤50 years old with Stage II (B2 or higher) or Stage III disease were assessed for CIN, telomere length and telomere maintenance mechanism (telomerase activation [TA]; alternative lengthening of telomeres [ALT]). Relative telomere length was measured by qPCR in somatic epithelial and cancer DNA. TA was measured with the TRAPeze assay, and tumors were evaluated for the presence of C-circles indicative of ALT. p53 mutation status was assessed in all available samples. DNA copy number changes were evaluated with Spectral Genomics aCGH.

Results

Tumors were classified as chromosomally stable (CIN-) and chromosomally instable (CIN+) by degree of DNA copy number changes. CIN- tumors (35%; n=6) had fewer copy number changes (<17% of their clones with DNA copy number changes) than CIN+ tumors (65%; n=13) which had high levels of copy number changes in 20% to 49% of clones. Telomere lengths were longer in CIN- compared to CIN+ tumors (p=0.0066) and in those in which telomerase was not activated (p=0.004). Tumors exhibiting activation of telomerase had shorter tumor telomeres (p=0.0040); and tended to be CIN+ (p=0.0949).

Conclusions

MSS rectal cancer appears to represent a heterogeneous group of tumors that may be categorized both on the basis of CIN status and telomere maintenance mechanism. MSS CIN- rectal cancers appear to have longer telomeres than those of MSS CIN+ rectal cancers and to utilize ALT rather than activation of telomerase.     

Patient Delay in Colorectal Cancer Patients: Associations with Rectal Bleeding and Thoughts about Cancer

Rectal bleeding is considered to be an alarm symptom of colorectal cancer. However, the symptom is seldom reported to the general practitioner and it is often assumed that patients assign the rectal bleeding to benign conditions. The aims of this questionnaire study were to examine whether rectal bleeding was associated with longer patient delays in colorectal cancer patients and whether rectal bleeding was associated with cancer worries. All incident colorectal cancer patients during a 1-year period in the County of Aarhus, Denmark, received a questionnaire. 136 colorectal cancer patients returned the questionnaire (response rate: 42%). Patient delay was assessed as the interval from first symptom to help-seeking and was reported by the patient. Patients with rectal bleeding (N = 81) reported longer patient intervals than patients without rectal bleeding when adjusting for confounders including other symptoms such as pain and changes in bowel habits (HR = 0.43; p = 0.004). Thoughts about cancer were not associated with the patient interval (HR = 1.05; p = 0.887), but more patients with rectal bleeding reported to have been wondering if their symptom(s) could be due to cancer than patients without rectal bleeding (chi² = 15.29; p<0.001). Conclusively, rectal bleeding was associated with long patient delays in colorectal cancer patients although more patients with rectal bleeding reported to have been wondering if their symptom(s) could be due to cancer than patients without rectal bleeding. This suggests that assignment of symptoms to benign conditions is not the only explanation of long patient delays in this patient group and that barriers for timely help-seeking should be examined.     

Heterogeneity of KRAS Mutation Status in Rectal Cancer

IntroductionAnti-EGFR targeted therapy is of increasing importance in advanced colorectal cancer and prior KRAS mutation testing is mandatory for therapy. However, at which occasions this should be performed is still under debate. We aimed to assess in patients with locally advanced rectal cancer whether there is intra-specimen KRAS heterogeneity prior to and upon preoperative chemoradiotherapy (CRT), and if there are any changes in KRAS mutation status due to this intervention.ResultsFor 20 (43%) out of the 47 patients, a KRAS mutation was detected. With 12 out of 20, the majority of these mutations affected codon 35. We did not obtained evidence that CRT results in changes of the KRAS mutation pattern. In addition, no intratumoral heterogeneity in the KRAS mutational status could be proven. This was true for both the biopsies prior to CRT and the resection specimens thereafter. The discrepancy observed in some samples when using the SNaPshot™ assay was due to insufficient sensitivity of this technique upon massive tumor regression by CRT as application of the therascreen® KRAS test revealed concordant results.ConclusionOur results indicate that the KRAS mutation status at the primary tumor site of rectal cancer is homogenous. Its assessment for therapeutic decisions is feasible in pre-therapeutic biopsies as well as in post-therapeutic resected specimens. The amount of viable tumor cells seems to be an important determinant for assay sensitivity and should thus be considered for selection of the analytical method.     

Impact of Diabetes on Oncologic Outcome of Colorectal Cancer Patients: Colon vs. Rectal Cancer

Background

To evaluate the impact of diabetes on outcomes in colorectal cancer patients and to examine whether this association varies by the location of tumor (colon vs. rectum).

Patients and methods

This study includes 4,131 stage I-III colorectal cancer patients, treated between 1995 and 2007 (12.5% diabetic, 53% colon, 47% rectal) in South Korea. Cox proportional hazards modeling was used to determine the prognostic influence of DM on survival endpoints.

Results

Colorectal cancer patients with DM had significantly worse disease-free survival (DFS) [hazard ratio (HR) 1.17, 95% confidence interval (CI): 1.00–1.37] compared with patients without DM. When considering colon and rectal cancer independently, DM was significantly associated with worse overall survival (OS) (HR: 1.46, 95% CI: 1.11–1.92), DFS (HR: 1.45, 95% CI: 1.15–1.84) and recurrence-free survival (RFS) (HR: 1.32, 95% CI: 0.98–1.76) in colon cancer patients. No association for OS, DFS or RFS was observed in rectal cancer patients. There was significant interaction of location of tumor (colon vs. rectal cancer) with DM on OS (P = 0.009) and DFS (P = 0.007).

Conclusions

This study suggests that DM negatively impacts survival outcomes of patients with colon cancer but not rectal cancer.     

体质指数与直肠癌的关系探讨

目的：探讨体质指数（bodymassindex,BMI）与我国直肠癌发病的关系,为直肠癌的预防提供参考。方法：用病例．对照研究方法分析353例首次确诊的直肠癌患者和354名健康人的BMI,比较两组人群BMI的情况。结果：首次确诊的直肠癌患者平均BMI为（24．54±4．48）kg／m^2,健康对照人群平均BMI为（23．58±3．12）kg／m^2,直肠癌患者的BMI明显高于健康对照人群,其差别具有统计学意义（P〈0．001）。根据性别的不同进行分组后,可以看出不同性别直肠癌患者的BMI均比健康对照组高。logistic回归分析,BMI的升高是直肠癌发生的危险因素,OR值为1．056（95％CI,1．027±1．089）。结论：直肠癌的发生与BMI有关。     

Efficiency of Non-Contrast-Enhanced Liver Imaging Sequences Added to Initial Rectal MRI in Rectal Cancer Patients

Purpose

The purpose of this study was to estimate the value of addition of liver imaging to initial rectal magnetic resonance imaging (MRI) for detection of liver metastasis and evaluate imaging predictors of a high risk of liver metastasis on rectal MRI.

Methods

We enrolled 144 patients who from October 2010 to May 2013 underwent rectal MRI with T2-weighted imaging (T2WI) and diffusion-weighted imaging (DWI) (b values = 50, 500, and 900 s/mm²) of the liver and abdominopelvic computed tomography (APCT) for the initial staging of rectal cancer. Two reviewers scored the possibility of liver metastasis on different sets of liver images (T2WI, DWI, and combined T2WI and DWI) and APCT and reached a conclusion by consensus for different analytic results. Imaging features from rectal MRI were also analyzed. The diagnostic performances of CT and an additional liver scan to detect liver metastasis were compared. Multivariate logistic regression to determine independent predictors of liver metastasis among rectal MRI features and tumor markers was performed. This retrospective study was approved by the Institutional Review Board, and the requirement for informed consent was waived.

Results

All sets of liver images were more effective than APCT for detecting liver metastasis, and DWI was the most effective. Perivascular stranding and anal sphincter invasion were statistically significant for liver metastasis (p = 0.0077 and p = 0.0471), while extramural vascular invasion based on MRI (mrEMVI) was marginally significant (p = 0.0534).

Conclusion

The addition of non-contrast-enhanced liver imaging, particularly DWI, to initial rectal MRI in rectal cancer patients could facilitate detection of liver metastasis without APCT. Perivascular stranding, anal sphincter invasion, and mrEMVI detected on rectal MRI were important imaging predictors of liver metastasis.     

直肠癌MRI研究进展

MRI是目前直肠癌诊断、分期的首选影像学方法。在判断肿瘤对邻近器官、结构的浸润程度上具有明显优势,尤其是对有较高复发风险的低位肿瘤。常规MRI尤其是高分辨MRI能够清晰显示直肠相关解剖,结合扩散加权成像（Diffusion weighted imaging,DWI）通过确定肿瘤边界,直肠系膜有无受侵,淋巴结及远处转移情况,可以准确有效的进行术前诊断、分期;DWI有助于鉴别辅助治疗后失活与存活组织、筛选出辅助治疗有效的患者,在评估治疗后疗效、提示患者预后方面发挥重要作用,也为临床制定治疗方案提供依据。同时也发现准确进行淋巴结分期、鉴别复发仍然存在困难,需要在以后进一步探讨,提高评估的准确性。本文就近年来MRI在直肠癌术前评价、术后疗效评估、复发监测及表观弥散系数（Apparent diffusion coefficient,ADC）的应用做一综述。     

Urinary Function following Laparoscopic Lymphadenectomy for Male Rectal Cancer

Objectives

Urinary function can be protected following open lateral node dissection (LND) with pelvic autonomic nerve preservation (PANP) for advanced rectal cancer. However data regarding urinary function after laparoscopic LND with PANP have not been reported. The goal of this study was to determine the effects of laparoscopic LND with PANP on urinary function in male patients with rectal cancer.

Methods

Urine flowmetry was performed using an Urodyn flowmeter. Patients were also asked to complete the standardized International Prostate Symptom Score (IPSS) questionnaire before surgery and 6 months after. In total, this study consisted of 60 males with advanced rectal cancer.

Results

No significant differences were seen in maximal urinary flow rate, voided volume or residual volume before and after surgery. The total IPSS score increased significantly after surgery and at least 41 patients (68.3%) reported there was no change in one of the seven IPSS questions.

Conclusions

Laparoscopic LND with PANP was relatively safe in preserving urinary function.     

直肠癌MRI研究进展

MRI是目前直肠癌诊断、分期的首选影像学方法。在判断肿瘤对邻近器官、结构的浸润程度上具有明显优势,尤其是对有较高复发风险的低位肿瘤。常规MRI尤其是高分辨MRI能够清晰显示直肠相关解剖,结合扩散加权成像(Diffusion weighted imaging,DWI)通过确定肿瘤边界,直肠系膜有无受侵,淋巴结及远处转移情况,可以准确有效的进行术前诊断、分期;DWI有助于鉴别辅助治疗后失活与存活组织、筛选出辅助治疗有效的患者,在评估治疗后疗效、提示患者预后方面发挥重要作用,也为临床制定治疗方案提供依据。同时也发现准确进行淋巴结分期、鉴别复发仍然存在困难,需要在以后进一步探讨,提高评估的准确性。本文就近年来MRI在直肠癌术前评价、术后疗效评估、复发监测及表观弥散系数(Apparent diffusion coefficient,ADC)的应用做一综述。     

Development of a Clinically-Precise Mouse Model of Rectal Cancer

Currently-used rodent tumor models, including transgenic tumor models, or subcutaneously growing tumors in mice, do not sufficiently represent clinical cancer. We report here development of methods to obtain a highly clinically-accurate rectal cancer model. This model was established by intrarectal transplantation of mouse rectal cancer cells, stably expressing green fluorescent protein (GFP), followed by disrupting the epithelial cell layer of the rectal mucosa by instilling an acetic acid solution. Early-stage tumor was detected in the rectal mucosa by 6 days after transplantation. The tumor then became invasive into the submucosal tissue. The tumor incidence was 100% and mean volume (±SD) was 1232.4 ± 994.7 mm³ at 4 weeks after transplantation detected by fluorescence imaging. Spontaneous lymph node metastasis and lung metastasis were also found approximately 4 weeks after transplantation in over 90% of mice. This rectal tumor model precisely mimics the natural history of rectal cancer and can be used to study early tumor development, metastasis, and discovery and evaluation of novel therapeutics for this treatment-resistant disease.     

STED Super-Resolution Microscopy of Clinical Paraffin-Embedded Human Rectal Cancer Tissue

Formalin fixed and paraffin-embedded human tissue resected during cancer surgery is indispensable for diagnostic and therapeutic purposes and represents a vast and largely unexploited resource for research. Optical microscopy of such specimen is curtailed by the diffraction-limited resolution of conventional optical microscopy. To overcome this limitation, we used STED super-resolution microscopy enabling optical resolution well below the diffraction barrier. We visualized nanoscale protein distributions in sections of well-annotated paraffin-embedded human rectal cancer tissue stored in a clinical repository. Using antisera against several mitochondrial proteins, STED microscopy revealed distinct sub-mitochondrial protein distributions, suggesting a high level of structural preservation. Analysis of human tissues stored for up to 17 years demonstrated that these samples were still amenable for super-resolution microscopy. STED microscopy of sections of HER2 positive rectal adenocarcinoma revealed details in the surface and intracellular HER2 distribution that were blurred in the corresponding conventional images, demonstrating the potential of super-resolution microscopy to explore the thus far largely untapped nanoscale regime in tissues stored in biorepositories.     

HEPIS的表达谱及其在直肠癌中的表达分析

HEPIS是一个新基因,关于HEPIS的表达和功能尚不十分清楚.本研究首先构建了HEPIS的表达载体pEGFP-C3-HEPIS,采用双荧光素酶报告实验检测了 HEPIS对Wnt、CRE、NF-KB和HRE信号通路的作用,随后采用RNA原位杂交(RISH)检测了 HEPIS在12种不同器官(乳腺,结肠,食道,肾,肝,肺,卵巢,胰腺,前列腺,直肠,胃和宫颈)的癌组织和正常组织的表达差异,并且检测了 HEPIS在40例直肠腺癌和直肠正常组织的表达.研究表明,在293T细胞转染pEGFP-C3-HEPIS后,无论是转录水平还是翻译水平,HEPIS均显著增加,且HEPIS可以显著抑制Wnt、CRE和NF-KB 3条信号通路,对NF-κB信号通路的抑制最为显著.RNA原位杂交实验证实HEPIS在12个器官的癌组织和正常组织中表达存在明显的表达差异.采用GEPIA对HEPIS在直肠癌及正常组织的表达差异进行了分析,结果表明HEPIS在正常直肠组织高表达,进一步采用RISH分析了 HEPIS在40例直肠癌和癌旁组织的表达差异,分析表明HEPIS在正常直肠组织的表达显著高于癌组织(P＜0.01),与GEPIA分析结果一致,说明HEPIS可能是直肠癌的一个潜在抑癌基因.     

Patient-Derived Organoids Predict Chemoradiation Responses of Locally Advanced Rectal Cancer

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The cluster-randomized Quality Initiative in Rectal Cancer trial: evaluating a quality-improvement strategy in surgery

Background

Following surgery for rectal cancer, two unfortunate outcomes for patients are permanent colostomy and local recurrence of cancer. We tested whether a quality-improvement strategy to change surgical practice would improve these outcomes.

Methods

Sixteen hospitals were cluster-randomized to the intervention (Quality Initiative in Rectal Cancer strategy) or control (normal practice) arm. Consecutive patients with primary rectal cancer were accrued from May 2002 to December 2004. Surgeons at hospitals in the intervention arm could voluntarily participate by attending workshops, using opinion leaders, inviting a study team surgeon to demonstrate optimal techniques of total mesorectal excision, completing postoperative questionnaires, and receiving audits and feedback. Main outcome measures were hospital rates of permanent colostomy and local recurrence of cancer.

Results

A total of 56 surgeons (n = 558 patients) participated in the intervention arm and 49 surgeons (n = 457 patients) in the control arm. The median follow-up of patients was 3.6 years. In the intervention arm, 70% of surgeons participated in workshops, 70% in intraoperative demonstrations and 71% in postoperative questionnaires. Surgeons who had an intraoperative demonstration provided care to 86% of the patients in the intervention arm. The rates of permanent colostomy were 39% in the intervention arm and 41% in the control arm (odds ratio [OR] 0.97, 95% confidence interval [CI] 0.63–1.48). The rates of local recurrence were 7% in the intervention arm and 6% in the control arm (OR 1.06, 95% CI 0.68–1.64).

Interpretation

Despite good participation by surgeons, the resource-intense quality-improvement strategy did not reduce hospital rates of permanent colostomy or local recurrence compared with usual practice. (ClinicalTrials.gov trial register no. {"type":"clinical-trial","attrs":{"text":"NCT00182130","term_id":"NCT00182130"}}NCT00182130.)Following surgery for rectal cancer, two unfortunate outcomes for patients are permanent colostomy and local recurrence of the cancer. Local recurrence is especially feared, because it is usually inoperable and patients can suffer a slow, painful death.¹ The use of total mesorectal excision, which involves dissection of the lymph node-bearing portion of the rectum,² has resulted in improved outcomes, with local recurrence rates as low as 1%–5% and rates of permanent colostomy of 10%–15%.^3–6 Population-based rates of local recurrence are unavailable for any North American jurisdiction, although a Canadian hospital series found that rates varied from 10% to 45% based on the practice volume and training of surgeons.⁷ A surgical report on health regions in the province of Ontario (population 13 million) found that rates of permanent colostomy varied from 31% to 41%.⁸ This geographic variation in outcomes, together with rates of inferior outcomes as compared to outcomes specific to total mesorectal excision, suggest that gaps exist in the quality of rectal surgery provided to patients with rectal cancer.Quality-improvement strategies for encouraging physicians to change practice include continuing medical education, the use of opinion leaders, and audit and feedback.^9–11 As well, improvement may be enhanced by using a participatory and supportive approach that focuses on the system and not on individual practitioners.^12,13 The small number of studies that have evaluated changes in surgeons’ practices often have targeted process measures, such as preoperative ordering of antibiotics, rather than patient outcomes, such as recurrence of cancer.^14,15We tested whether use of a surgeon-directed quality-improvement strategy would improve hospital rates of permanent colostomy and local recurrence of cancer among patients undergoing surgery for rectal cancer. We used the Quality Initiative in Rectal Cancer (QIRC) strategy, which integrates quality-improvement interventions and principles to encourage surgeons to provide optimal total mesorectal excision to patients with rectal cancer.¹⁶     

Differences in Survival between Colon and Rectal Cancer from SEER Data

Background

Little is known about colorectal cancer or colon and rectal cancer. Are they the same disease or different diseases?

Objectives

The aim of this epidemiology study was to compare the features of colon and rectal cancer by using recent national cancer surveillance data.

Design and setting

Data included colorectal cancer (1995–2008) from the Surveillance, Epidemiology, and End Results Program (SEER) database. Only adenocarcinoma was included for analysis.

Patients

A total of 372,130 patients with a median follow-up of 32 months were analyzed.

Main outcome measures

Mean survival of patients with the same stage of colon and rectal cancer was evaluated.

Results

Around 35% of patients had stage information. Among them, colon cancer patients had better survival than those with rectal cancer, by a margin of 4 months in stage IIB. In stage IIIC and stage IV, rectal cancer patients had better survival than colon cancer patients, by about 3 months. Stage IIB colorectal cancer patients had a poorer prognosis than those with stage IIIA and IIIB colorectal cancer. After adjustment of age, sex and race, colon cancer patients had better survival than rectal cancer of stage IIB, but in stage IIIC and IV, rectal cancer patients had better survival than colon cancer.

Limitations

The study is limited by its retrospective nature.

Conclusion

This was a population-based study. The prognosis of rectal cancer was not worse than that of colon cancer. Local advanced colorectal cancer had a poorer prognosis than local regional lymph node metastasis. Stage IIB might require more aggressive chemotherapy, and no less than that for stage III.     

肛提肌外腹会阴联合切除术治疗低位进展期直肠癌的临床研究

目的:探讨肛提肌外腹会阴联合切除术(ELAPE)在低位进展期直肠癌中的应用。方法:回顾性分析2011年1月至2013年12月我院胃肠外科30例接受ELAPE和18例接受传统腹会阴联合直肠癌切除术(APR)的低位直肠癌病人的临床资料,通过对手术用时、失血量、术中穿孔发生率、术后住院时间、术后并发症以及出院后随访相关指标等的比较来分析ELAPE的可行性。结果:和APR相比,ELAPE治疗低位直肠癌可降低术中直肠穿孔和CRM阳性的发生率,术后会阴部切口愈合延迟率以及出院后肿瘤局部复发、远处转移以及患者死亡的发生率,差异均有统计学意义(P0.05)。行ELAPE术的手术时间、术中出血量、平均住院天数,术后引流管拔管时间、尿潴留发生率以及出院随访的骶尾部不适率均稍大于行APR术的患者,但差异均尚无统计学意义(P0.05);进行两种手术的患者住院两组间围手术期死亡病例、会阴部血清肿的发生率、肠梗阻发生率、造口问题发生率以及出现并发症的情况差异不大(P0.05)。结论:和APR相比,ELAPE治疗低位进展期直肠癌可降低术中肠管穿孔发生率、CRM阳性率和住院期间发生会阴部切口愈合延迟率,短期随访预后良好,有望成为治疗进展期低位直肠癌的推荐术式。