Circulating plasma microRNAs in colorectal neoplasia: A pilot study in assessing response to therapy

IntroductionCurrent serological surveillance markers to monitor colorectal cancer (CRC) or colorectal advanced adenomas (CAA) are hampered by poor sensitivity and specificity. The aim of this study is to identify and validate a panel of plasma microRNAs which change in expression after resection of such lesions.MethodsA prospectively maintained colorectal surgery database was queried for patients in whom both pre- and post-procedural serum samples had been obtained. An initial screening analysis of CRC and CAA patients (5 each) was conducted using screening cards for 380 miRNAs. Four identified miRNAs were combined with a previously described panel of 7 miRNAs that were diagnostically predictive of CRC and CAA. Differential miRNA expression was assessed using quantitative real-time polymerase chain reaction(qRT-PCR).ResultsFifty patients were included (n = 27 CRC, n = 23 CAA). There was no difference in age, gender, or race profile of CRC patients compared to CAA patients. Six miRNA were significantly increased after CRC resection (miR-324, let7b, miR-454, miR-374a, miR-122, miR-19b, all p<0.05), while three miRNAs were significantly increased following CAA resection (miR-454, miR-374a, miR-122, all p<0.05). Three miRNA were increased in common for both (miR-454, miR-374a, miR-122).DiscussionThe expression of miRNAs associated with neoplasia (either CRC or CAA) was significantly increased following surgical resection or endoscopic removal of CRC or CAA. Future studies should focus on the evaluation of these miRNAs in CRC and CAA prognosis.     

miR-33a-5p in small extracellular vesicles as non-invasive biomarker for oxaliplatin sensitivity in human colorectal cancer cells

microRNAs (miRNAs) contained in small extracellular vesicles (sEVs) are candidates for non-invasive biomarkers. Oxaliplatin (L-OHP) has been approved for advanced colorectal cancer (CRC) chemotherapy. However, the response to L-OHP differs among CRC patients. In addition, CRC cells often acquire the resistance to L-OHP. This study aimed at the prediction of L-OHP sensitivity by measuring extracellular miRNAs levels. Firstly, we compared intracellular miRNAs expressions in L-OHP-sensitive CRC cells (SW620 and HCT116 cells) with those in acquired and intrinsic L-OHP-resistant cells. In microarray and real-time RT-PCR analyses, the intracellular miR-33a-5p, miR-210–3p, and miR-224–5p expressions were lower in acquired and intrinsic L-OHP-resistant CRC cells than sensitive cells. Furthermore, in SW620 cells, L-OHP sensitivity was decreased by miR-33a-5p inhibitor. On the other hand, miR-210–3p or miR-224–5p inhibitor did not affect L-OHP sensitivity in SW620 cells. Secondly, the amount of miR-33a-5p, miR-210–3p, and miR-224–5p in sEVs was compared. The amount of miR-33a-5p and miR-210–3p in sEVs secreted from acquired and intrinsic L-OHP-resistant cells tended to be small. miR-224–5p was not detected in sEVs secreted from three types of CRC cells examined. To the best of our knowledge, this is the first study demonstrating that miR-33a-5p and/or miR-210–3p in sEVs would be candidates for biomarkers of L-OHP sensitivity. In particular, miR-33a-5p is a promising candidate because it would be directly involved in L-OHP sensitivity.     

Bulky DNA adduct levels in normal-appearing colon mucosa,and the prevalence of colorectal adenomas

Abstract

Purpose: Examine the association between bulky DNA adduct levels in colon mucosa and colorectal adenoma prevalence, and explore the correlation between adduct levels in leukocytes and colon tissue.

Methods: Bulky DNA adduct levels were measured using ³²P-postlabelling in biopsies of normal-appearing colon tissue and blood donated by 202 patients. Multivariable logistic regression was used to examine associations between DNA adducts, and interactions of DNA adduct-DNA repair polymorphisms, with the prevalence of colorectal adenomas. Correlation between blood and tissue levels of DNA adducts was evaluated using Spearman’s correlation coefficient.

Results: An interaction between bulky DNA adduct levels and XPA rs1800975 on prevalence of colorectal adenoma was observed. Among individuals with lower DNA repair activity, increased DNA adduct levels were associated with increased colorectal adenoma prevalence (OR?=?1.41 per SD increase, 95%CI: 0.92–2.18). Conversely, among individuals with normal DNA activity, an inverse association was observed (OR?=?0.60 per SD increase, 95%CI: 0.34–1.07). Blood and colon DNA adduct levels were inversely correlated (ρ?=??0.20).

Conclusions: Among genetically susceptible individuals, higher bulky DNA adducts in the colon was associated with the prevalence of colorectal adenomas. The inverse correlation between blood and colon tissue measures demonstrates the importance of quantifying biomarkers in target tissues.     

Impact of hyperthermic intraperitoneal chemotherapy on Hsp27 protein expression in serum of patients with peritoneal carcinomatosis

Despite the strong rationale for combining cytoreductive surgery (CRS) with hyperthermic intraperitoneal chemotherapy (HIPEC) in patients with peritoneal carcinomatosis, thermotolerance and chemoresistance might result from heat shock protein overexpression. The aim of the present study was thus to determine whether the heat shock protein 27 (Hsp27), a potential factor in resistance to treatment, could have a higher level in serum from patients under this combined therapy. Patients receiving CRS plus HIPEC for peritoneal carcinomatosis (group 1), patients with cancer or a history of cancer undergoing abdominal surgery (group 2), and patients without malignancies undergoing abdominal surgery (group 3) were included. Hsp27 serum levels were determined before and at different times following CRS and HIPEC using enzyme-linked immunosorbent assay. In group 1 (n = 25), the high Hsp27 levels, observed at the end of surgery compared with before (p < 0.0001), decreased during HIPEC, but remained significantly higher than before surgery (p < 0.0005). In groups 2 (n = 11) and 3 (n = 15), surgery did not significantly increase Hsp27 levels. A targeted molecular strategy, inhibiting Hsp27 expression in tumor tissue, could significantly reduce resistance to the combined CRS plus HIPEC treatment. This approach should be further assessed in a clinical phase I trial.     

结直肠癌淋巴结微转移灶的临床病理研究

摘要目的：探讨结直肠癌淋巴结微转移灶的特点及其临床病理意义。方法：对我院2001年1月-2006年10月收治的56例无组织学淋巴结转移的结直肠癌患者的650个淋巴结进行微转移灶的免疫组织化学和组织学检测,并分析微转移灶与患者临床病理特征之间的关系。结果-通过对41,zm厚淋巴结连续切片进行观察,连续切片五片的微转移灶检出率较一片和两片的的微转移灶检出率显著升高（P〈0．05）。按解剖部位分组的患者微转移灶阳性率1组有66．67％（16／24）,2组有60％（9／15）,3组有41．2％（7／17）,无显著性差别（P〉0．05）。微转移灶与无组织学淋巴结转移的结直肠癌患者的肿瘤浸润深度（P=0．013）和肿块大小（P=0．037）存在显著性相关,但有微转移灶和无微转移灶的无组织学淋巴结转移的结直肠癌患者总体生存状况比较无显著差异（P〉0．05）。结论：微转移灶在无组织学淋巴结转移的结直肠癌淋巴结中较常见,与肿瘤大小和浸润深度有关,但与患者的预后无显著相关性。     

血清CA153、TK1、TSGF联合检测对乳腺癌诊断的价值

目的：探讨细胞表面糖蛋白（CA153）、胸苷激酶（TKl）、肿瘤生长因子（TSGF）等联合检测在乳腺癌诊断中的应用价值。方法：73例确诊为乳腺癌患者的血清标本作为实验组;66例乳腺良性疾病（包括乳腺纤维瘤、囊肿、增生等）及50例健康人群血清标本作为对照组。采用电化学发光法检测血清CAl53,免疫化学发光法检测TKl,化学显示法检测TSGF的表达。结果：血清CAl53、TKl及TSGF对乳腺癌的敏感性分别为54．8％、53．4％及79．5％,特异性分别为87．9％、81．8％及83．3％;血清CAl53、TKl、TSGF联合检测乳腺癌的敏感性为89．0％,特异性为92．4％。结论：与单项指标检测相比,多个指标联合检测提高了对乳腺癌早期诊断的敏感性．同时又有较好的特异性．有助于良、恶乳腺疾病的鉴别．具有一定的临床应用价值．     

结直肠癌组织异常表达miRNA的鉴定

目的：筛选结直肠癌组织异常表达的miRNAs。方法：采用Agilem基因芯片（V12．0）分析结直肠癌组织及其配对正常粘膜组织间差异表达的miRNAs,MiRNAs错误发生率（FDR）〈0．05和微矩阵显著性分析（SAM）q值〈0．05为差异显著。结果：鉴定出结直肠癌中32个差异表达的miRNAs,显著上调和下调各16个。实时定量PCR（RT—qPCR）证实基因芯片中4个表达上调的miRNAs在结直肠癌组织中也显著上调。结论：MiRNA基因芯片鉴定出了结直肠癌组织一系列新的差异表达的miRNAs。     

恶性滋养细胞肿瘤治疗、预后的评价-附169例临床分析

目的:对恶性滋养细胞肿瘤按2000年国际妇产科联盟(FIGO)预后评分标准评分并选择治疗方案的疗效进行评价.方法:恶性滋养细胞肿瘤患者169例,治疗前按2000年FIGO预后评分标准进行分期及评分,以此选择治疗方案.结果:169例患者中,绒毛膜癌49例,30例为高危,19例为低危;侵蚀性葡萄胎120例,8例为高危,112例为低危.治疗原则:低危患者以单药化疗为主,高危患者采用多药联合化疗.本组患者的1年、3年、5年生存率分别为98.8％、97.6％及96.9％.无一例因毒副反应或并发症而死亡.结论:治疗前应用2000年FIGO顸后评分系统选择恶性滋养细胞肿瘤治疗方案取得较好的临床效果,新分期系统对指导临床治疗有重大意义.     

局部进展期胃癌术前动静脉结合化疗的疗效观察

目的:局部进展期胃癌,即使没有出现远处转移或腹膜种植,预后也极不理想.我们对局部进展期胃癌患者进行术前mECF方案的动静脉结合化疗,以评价其治疗疗效.方法:选取自2009年1月至2011年6月间我院局部进展期胃癌Ⅱ-ⅢC期患者38例,均由影像学确定淋巴结高度可疑转移或浸润、包绕主要血管结构,且没有发现远处转移或腹膜种植,进行2个周期的术前动静脉结合化疗后,行手术治疗.记录化疗毒性反应、临床、病理缓解率、手术并发症发生率和病死率以及1年无病生存率.结果:化疗毒性反应低,3级反应不超过10％,仅有1例出现4级反应,表现为腹泻、恶心和呕吐.38例患者中临床CR有6例,占15.8％,PR17例,占44.7％,NC13例(34.2％),PD2例(5.3％),RR为60.5％(23/38).全部患者均施行了手术,37例患者进行了根治性手术,R0切除率为97％.病理缓解率为5例完全缓解(13.2％),21例部分缓解(55.3％),8例轻微缓解(21.1％),4例未缓解(10.5％).手术并发症发生率为7.9％,无治疗相关死亡发生.1年无病生存率为81.6％.结论:局部进展期胃癌患者术前进行mECF方案的动静脉化疗毒性反应低,疗效理想,之后行手术完整切除后临床效果突出,是理想的治疗方法.     

SNAIL- and SLUG-induced side population phenotype of HCT116 human colorectal cancer cells and its regulation by BET inhibitors

Epithelial-mesenchymal transition (EMT) is associated with cancer malignancies such as invasion, metastasis, and drug resistance. In this study, HCT116 human colorectal cancer cells were transduced with SLUG or SNAIL retroviruses, and EMT cells with mesenchymal morphology were established. The EMT cells showed a high invasive activity and resistance to several anticancer agents such as methotrexate, SN-38, and cisplatin. Furthermore, they contained about 1–10% side population (SP) cells that were not stained by Hoechst 33342. This SP phenotype was not stable; the isolated SP cells generated both SP and non-SP cells, suggesting a potential for differentiation. Gene expression analysis of SP cells suggested the alteration of genes that are involved in epigenetic changes. Therefore, we examined the effect of 74 epigenetic inhibitors, and found that two inhibitors, namely I-BET151 and bromosporine, targeting the bromodomain and extra-terminal motif (BET) proteins, decreased the ratio of SP cells to <50% compared with the control, without affecting the immediate efflux of Hoechst 33342 by transporters. In addition, compared with the parental cells, the EMT cells showed a higher sensitivity to I-BET151 and bromosporine. This study suggests that EMT development and SP phenotype can be independent events but both are regulated by BET inhibitors in SLUG- or SNAIL-transducted HCT116 cells.