Proteins of the retinoblastoma pathway,FEN1 and MGMT are novel potential prognostic biomarkers in pancreatic adenocarcinoma

Background

We studied the expression of some major proteins involved in cell-cycle regulation and DNA repair, the roles of which are not well known in pancreatic ductal adenocarcinoma (PDAC), but which have a significant impact on carcinogenesis of many other cancers.

Molecular correlates with MGMT promoter methylation and silencing support CpG island methylator phenotype-low (CIMP-low) in colorectal cancer

Background

The CpG island methylator phenotype (CIMP or CIMP‐high) with widespread promoter methylation is a distinct epigenetic phenotype in colorectal cancer. In contrast, a phenotype with less widespread promoter methylation (CIMP‐low) has not been well characterised. O‐6‐methylguanine‐DNA methyltransferase (MGMT) promoter methylation and silencing have been associated with G>A mutations and microsatellite instability‐low (MSI‐low).

Aim

To examine molecular correlates with MGMT methylation/silencing in colorectal cancer.

Methods

Utilising MethyLight technology, we quantified DNA methylation in MGMT and eight other markers (a CIMP‐diagnostic panel; CACNA1G, CDKN2A (p16), CRABP1, IGF2, MLH1, NEUROG1, RUNX3 and SOCS1) in 920 population‐based colorectal cancers.

Results

Tumours with both MGMT methylation and loss were correlated positively with MSI‐low (p = 0.02), CIMP‐high (⩾6/8 methylated CIMP markers, p = 0.005), CIMP‐low (1/8–5/8 methylated CIMP markers, p = 0.002, compared to CIMP‐0 with 0/8 methylated markers), KRAS G>A mutation (p = 0.02), and inversely with 18q loss of heterozygosity (p = 0.0002). Tumours were classified into nine MSI/CIMP subtypes. Among the CIMP‐low group, tumours with both MGMT methylation and loss were far more frequent in MSI‐low tumours (67%, 12/18) than MSI‐high tumours (5.6%, 1/18; p = 0.0003) and microsatellite stable (MSS) tumours (33%, 52/160; p = 0.008). However, no such relationship was observed among the CIMP‐high or CIMP‐0 groups.

Conclusion

The relationship between MGMT methylation/silencing and MSI‐low is limited to only CIMP‐low tumours, supporting the suggestion that CIMP‐low in colorectal cancer may be a different molecular phenotype from CIMP‐high and CIMP‐0. Our data support a molecular difference between MSI‐low and MSS in colorectal cancer, and a possible link between CIMP‐low, MSI‐low, MGMT methylation/loss and KRAS mutation.     

Prognostic significance of DNA repair proteins MLH1, MSH2 and MGMT expression in non-small-cell lung cancer and precursor lesions

Aims:  To investigate the role of DNA repair proteins and their prognostic significance in non-small-cell lung cancer (NSCLC).
Methods and results:  A retrospective analysis of 108 cases of stage I–II NSCLC was undertaken. Immunohistochemical expression of DNA repair proteins MLH1, MSH2 and MGMT was assessed using tissue microarrays of paraffin-embedded samples of invasive carcinoma and precursor lesions. Results were analysed in relation to clinicopathological parameters and patient survival. Reduced expression of MLH1 was found in 58.5% of tumours and occurred less frequently in poorly differentiated tumours ( P  = 0.044) and large cell carcinomas ( P  = 0.004). MSH2 and MGMT expression was reduced in 18.1% and 77.8% of cases, respectively. There was an inverse relationship between MLH1 and MSH2 expression ( P  = 0.012). Normal expression of MLH1, MSH2 and MGMT was found in all cases of squamous metaplasia and squamous dysplasia. Only a single case of carcinoma in situ (12.5%) showed reduced MLH1, none showed reduced MSH2 and 25% showed reduced MGMT. Survival analyses showed no prognostic significance based on expression of MLH1 ( P  = 0.92), MSH2 ( P  = 0.78) or MGMT ( P  = 0.57).
Conclusions:  Reduction in expression of DNA repair proteins MLH1, MSH2 and MGMT is relatively common in NSCLC, appears to be a late event in the development of invasive malignancy and does not influence survival in this patient cohort.     

MGMT基因启动子甲基化检测在脑胶质瘤化疗中的意义

背景与目的：如何预测和克服肿瘤细胞对化疗药物的耐药性,实施个体化治疗是肿瘤化疗急需解决的问题。与基因启动子甲基化密切相关的DNA损伤修复基因O^6-甲基鸟嘌呤-DNA甲基转移酶（O^6-methylguanine-DNA methyhransferase,MGMT）表观沉默与肿瘤对烷化剂药物化疗敏感性密切相关。本研究探讨检测MGMT基因启动子CpG岛甲基化在判断脑胶质瘤患者预后及预测肿瘤对烷化剂药物耐药性中的意义。方法：甲基化特异性PCR（MSP）法检测脑胶质瘤组织及肿瘤细胞株MGMT基因启动子甲基化状态,蛋白印迹和免疫组化法测定蛋白表达。MTF法检测肿瘤细胞株对烷化剂药物敏感性,将患者随访资料针对MGMT甲基化状态绘制Kaplan-Meier生存曲线,并进行log—rank检验分析。结果：39例脑胶质瘤患者组织MGMT基因启动子甲基化发生率为46．2％,蛋白表达阳性率为61．5％,且肿瘤组织中MGMT基因甲基化状态与蛋白表达显著相关（P〈0．05）：6例正常组织均未检测出基因甲基化。MGMT基因过甲基化的脑胶质瘤SHG44细胞株用5-Aza-CdR处理后完全脱甲基化．MGMT蛋白恢复了表达,同时细胞株对烷化剂药物敏感性也发生逆转．由敏感转变为耐受。在采用手术、放疗和烷化剂尼莫司汀化疗等综合治疗的39例脑胶质瘤患者中,MGMT基因甲基化的患者生存率显著高于MGMT基因未甲基化患者（P〈0．05）。结论：MGMT基因甲基化状态与蛋白表达及肿瘤细胞对烷化剂药物敏感性密切相关,有可能替代MGMT蛋白检测成为判断脑胶质瘤患者预后和预测肿瘤对烷化剂化疗耐药性的标志分子。