Cerivastatin enhances the cytotoxicity of 5-fluorouracil on chemosensitive and resistant colorectal cancer cell lines

Cerivastatin is one of the synthetic 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors used for the treatment and prevention of hypercholesterolaemia. The observation that patients receiving this drug had a lower incidence at cancer led to our interest in using it as a putative anticancer agent. In this study, we tested the cytotoxicity of cerivastatin on a panel of 5-fluorouracil (5FU) sensitive and resistant cell lines in vitro. Cerivastatin was cytotoxic to both 5FU sensitive and resistant cells. Cerivastatin significantly augmented the cytotoxic effect of 5FU on drug sensitive (6-22-fold) and resistant (229-310-fold) cell lines. Cerivastatin and 5FU acted synergistically. Cerivastatin inhibited nuclear factor kappaB DNA binding activity. The enhancing effect of cerivastatin on 5FU was partially mevalonate pathway independent. Cerivastatin may allow successful 5FU therapy in chemoresistant patients.     

The disturbances of the zinc concentration in the blood in selected neoplasms

A study was carried out on 92 patients (58 males and 34 females) aged 42–76 treated for malignant neoplasm of the gastrointestinal tract (54 patients with colorectal carcinoma, 38 with gastric carcinoma). In all patients, the zinc serum concentration was measured and the results obtained were referred to some epidemiological-clinical factors (sex, age, primary cause of cancer, the stage of clinical progression, and histological type). The results showed that the most pronounced hypozincemia occurred in male patients with mucous membrane carcinoma of the stomach.     

Phase I trial of interleukin-2 and high-dose arginine butyrate in metastatic colorectal cancer

Interleukin-2 (IL-2) and sodium butyrate allow rats to be cured of peritoneal carcinomatosis from colon cancer. We performed a phase I trial of IL-2 and high-dose arginine butyrate (ArgB) in patients with advanced metastatic colorectal cancer. Patients and methods: From April to July 1997, six patients were included in the trail; they had a median age of 52 years, four had a performance status of 0, two had a performance status of 1 with normal biological functions. All patients had received at least two prior lines of chemotherapy. A fixed dose of 18 MIU/m² IL-2,was administered by subcutaneous injection and ArgB was delivered via continuous intravenous infusion on days 1–6 with escalating doses starting at 2 g kg⁻¹ day⁻¹. Results: The planned dose escalation was not possible because of toxicities. A daily ArgB dose of 2 g/kg was delivered for nine cycles. Level 2 (4 g/kg) could not be delivered in three of the six patients because of liver toxicity. The dose-limiting toxicities were fatigue and liver function disturbances. The maximum tolerated dose for ArgB was 3 g kg⁻¹ day⁻¹, in combination with IL-2 at 12 MIU m² day⁻¹. No clinical response was seen. Pharmacokinetic analysis showed large intra- and interindividual variations. Conclusion: This schedule with a high dose of ArgB proved to be highly toxic with liver insufficiency. We will be running another trial with lower doses of ArgB calculated from the schedule used in the experimental model, starting at a dose of 20 mg kg⁻¹ day⁻¹ for ArgB and 200 000 UI kg⁻¹ day⁻¹ IL-2, every 8 h. Received: 13 May 1999 / Accepted: 28 October 1999     

Cancer vaccines: single-epitope anti-idiotype vaccine versus multiple-epitope antigen vaccine

In this study, we compared the immunogenicity and tumor-protective activity of anti-idiotypic antibodies mimicking a single tumor-associated epitope and tumor-associated antigen expressing multiple potentially immunogenic epitopes. We focused our study on the colorectal-carcinoma(CRC)-associated antigen GA733 (also known as CO17-1A/KS1-4/KSA/EpCAM). Monoclonal anti-idiotypic antibody (Ab2) BR3E4 was produced against murine anti-CRC mAb CO17-1A (Ab1) in rats. Full-length native GA733 protein was isolated from human tumor cells, and the extracellular domain protein (GA733-2E) was isolated from supernatants of recombinant baculovirus-infected insect cells by immunoafffinity chromatography. The immunomodulatory activity of the Ab2 was compared with that of the antigen, both in rabbits and in mice. Mice, like humans but not rabbits, express a GA733 antigen homologue on some of their normal tissues. Thus, these in vivo models allow the comparison of the immunogenicity of Ab2 and antigen in the presence (mice) and absence (rabbits) of normal tissue expression and immunological tolerance of the GA733 antigen homologue. In rabbits, aluminum-hydroxide(alum)-precipitated native GA733 antigen was superior to alum-precipitated Ab2 in inducing specific humoral immunity. In mice, alum-precipitated recombinant GA733-2E antigen, but not alum-precipitated Ab2, induced specific humoral immunity. However, when the Ab2 was administered to mice in Freund's complete adjuvant, specific humoral immune responses were elicited. Ab2 in complete Freund's adjuvant and GA733-2E in alum were compared for their capacity to induce antigen-specific cellular immunity in mice. Whereas lymphoproliferative responses were obtained with the recombinant antigen only, delayed-type hypersensitivity responses were obtained with both recombinant antigen and Ab2, although these responses were lower than after antigen immunization. The recombinant antigen in alum did not protect mice against challenge with antigen-positive syngeneic murine CRC cells. Similar studies with Ab2 BR3E4 mimicking the CO17-1A epitope were not possible because the tumor cells do not express this epitope after transfection with the human GA733-2 cDNA. However, similar studies with Ab2 mimicking the epitope defined by mAb GA733, which is expressed by the transfected tumor cells, indicated a lack of tumor-protective activity of this Ab2. In contrast, the full-length antigen expressed by recombinant adenovirus inhibited the growth of established tumors in mice. In conclusion, soluble antigen is a more potent modulator of humoral and cellular immune responses than Ab2, both administered in adjuvant. However, for induction of protective immunity, the immunogenicity of the antigen must be further enhanced, e.g., by expression of the antigen in a viral vector. Received: 27 December 1999 / Accepted: 27 January 2000     

应用PCR产物直接银染测序技术检测大肠癌p53基因点突变

应用PCR-SSCP结合PCR产物直接银染测序技术对24例大肠癌p53基因第5-7外显子进行点突变的研究。结果检出5例(26.7%)阳性,均为错义突变;其中3例为碱基GC到 AT的转换, 1例为GC到TA的颠换,另1例为AT到CG的颠换,后者尚未见报道。突变位点分布在p53基因第141、175、245、248和258位密码子,其中4例发生在CpG位点。本文对p53基因点突变谱的分析为大肠癌的病因学研究提供了科学依据, 并讨论了PCR产物直接银染测序技术的优越性。 Abstract:Mutations in exon 5~7 of p53 were screened in 24 cases of colorectal carcinoma by a combination of PCR-SSCP and PCR-product DNA silver sequencing.The results showed that all 5(26.7%) cases of point mutations detected were missense mutations,including 3 cases of GC to AT transitions,1 case of GC to TA transversion and another case of AT to CG transversion.The latter has not been reported before.The mutations occurred at codons 141,175,245,248 and 258 respectively,and 4 cases of these five mutations occurred at CpG dinucleotides.The analysis of p53 mutation spectra can provide clues to the etiology of colorectal carcinoma.The advantages of DNA silver sequencing are also discussed.     

结直肠癌组织miR-137-3p、miR-410-3p表达水平与上皮间充质转化和预后的关系分析

摘要 目的：探讨结直肠癌（CRC）组织微小RNA-137-3p（miR-137-3p）、微小RNA-410-3p（miR-410-3p）的表达与上皮间充质转化（EMT）和预后的关系。方法：选取2017年2月至2019年2月本院收治的115例接受CRC根治术治疗的患者，采用实时荧光定量聚合酶链式反应（qRT-PCR）检测癌组织及癌旁组织中miR-137-3p、miR-410-3p表达、EMT标志物E-钙粘蛋白（E-cadherin）、波形蛋白（Vimentin）的mRNA表达并进行相关性分析，分析癌组织中miR-137-3p、miR-410-3p表达与CRC临床病理特征的关系。术后随访3年，采用Kaplan-Meier法分析miR-137-3p、miR-410-3p高表达和低表达患者的预后情况。结果：癌组织中miR-137-3p表达及E-cadherin mRNA表达水平低于癌旁组织，miR-410-3p表达及Vimentin mRNA表达水平高于癌旁组织（P＜0.05）。癌组织中miR-137-3p表达与E-cadherin mRNA表达水平、miR-410-3p表达与Vimentin mRNA表达水平分别呈正相关（P＜0.05），癌组织中miR-137-3p表达与Vimentin mRNA表达水平、miR-410-3p表达与E-cadherin mRNA表达水平分别呈负相关（P＜0.05）。miR-137-3p、miR-410-3p表达与TNM分期、肿瘤分化程度、淋巴结转移有关（P＜0.05）。Kaplan-Meier生存曲线分析显示，miR-137-3p低表达、miR-410-3p高表达患者3年累积生存率下降（P＜0.05）。结论：CRC组织中miR-137-3p表达下调、miR-410-3p表达上调，miR-137-3p、miR-410-3p表达水平与EMT密切相关，且miR-137-3p高表达、miR-410-3p低表达患者的预后更好。     

Analysis of colorectal cancer glyco‐secretome identifies laminin β‐1 (LAMB1) as a potential serological biomarker for colorectal cancer

The high mortality rate in colorectal cancer is mostly ascribed to metastasis, but the only clinical biomarker available for disease monitoring and prognosis is the carcinoembryonic antigen (CEA). However, the prognostic utility of CEA remains controversial. In an effort to identify novel biomarkers that could be potentially translated for clinical use, we collected the secretomes from the colon adenocarcinoma cell line HCT‐116 and its metastatic derivative, E1, using the hollow fiber culture system, and utilized the multilectin affinity chromatography approach to enrich for the secreted glycoproteins (glyco‐secretome). The HCT‐116 and E1 glyco‐secretomes were compared using the label‐free quantitative SWATH‐MS technology, and a total of 149 glycoproteins were differentially secreted in E1 cells. Among these glycoproteins, laminin β‐1 (LAMB1), a glycoprotein not previously known to be secreted in colorectal cancer cells, was observed to be oversecreted in E1 cells. In addition, we showed that LAMB1 levels were significantly higher in colorectal cancer patient serum samples as compared to healthy controls when measured using ELISA. ROC analyses indicated that LAMB1 performed better than CEA at discriminating between colorectal cancer patients from controls. Moreover, the diagnostic performance was further improved when LAMB1 was used in combination with CEA.     

MUC1对人结肠癌细胞HCT116生物学行为的影响

目的:观察MUC1对人结肠癌细胞HCT116增殖、侵袭及化疗敏感性的影响。方法:采用MUC1表达阴性的结肠癌细胞株HCT116,通过慢病毒转染、嘌呤霉素筛选、半定量RT-PCR和Western blot鉴定构建稳定表达MUC1的HCT116细胞株;实验分空病毒组和MUC1病毒组;CCK实验和软琼脂克隆形成实验检测两组细胞的增殖能力,Transwell侵袭实验检测两组细胞的侵袭能力;MTT法和流式细胞仪检测两组细胞对奥沙利铂的敏感性,酶底物法检测Caspase-3活性。结果:获得稳定表达MUC1的HCT116细胞株;两组细胞贴壁生长无差异;与空病毒组相比,MUC1病毒组细胞软克隆形成数增加,穿过小室的细胞数增加(P0.05);MUC1病毒组细胞对奥沙利铂的敏感性降低,MUC1病毒组Caspase-3活性水平低于空病毒组(P0.05)。结论:MUC1与结肠癌的非锚定依赖生长、侵袭和化疗敏感性有关。     

腹腔镜监控肠镜下（LMCP）息肉摘除术对比常规肠镜下息肉切除的随机对照临床研究

目的:探讨腹腔镜监控下的肠息肉摘除术(LMCP)治疗效果,比较LMCP肠息肉摘除术对比常规肠镜下息肉切除的临床治疗效果及预后情况。方法:将符合条件的所有手术患者随机分为两组,每组各41例,其中试验组使用腹腔镜监控下结肠镜息肉摘除术,对照组单纯使用肠镜行息肉切除术。所有病人均观察并记录其预后情况。结果:研究共纳入82例病人,男53例,女29例,平均年龄70岁。息肉平均大小为2.0 cm。所有患者术后无并发症。试验组和对照组的第一次通便时间分别为13.2 h和24.5h,统计学具有显著性差异P<0.001,风险比为1.81,95%置信区间为[1.13-3.00]。试验组和对照组的总住院天数分别为4.5天和8.0天,统计学具有显著性差异P<0.001,风险比为4.15,置信区间95%CI为[2.40-7.18]。结论:LMCP术对病人具有显著的获益,可以避免不必要的并发症,手术操作更安全。因此,LMCP是一种安全有效的方法,并且创伤更小,住院周期更短,是息肉切除术首选的方法。