Weekly 5-fluorouracil combined with PALA: toxic and therapeutic effects in colorectal cancer

A total of 51 patients (eight previously treated) received PALA added to 5-fluorouracil (5FU) given weekly. After 32 patients, the PALA schedule was changed from every other week to weekly, 24 hours preceding 5FU in accordance with preclinical leads (see text). Both schedules were associated with moderately severe toxic effects related primarily to PALA (skin rash) or to the combined effects of both drugs (diarrhea, vomiting, conjunctivitis, and neurotoxicity). Overall nine partial responses were observed, including three in patients previously treated with 5FU. However, future studies with this combination utilizing the current or other previously published schedules are not warranted in colorectal cancer. Since toxicity is a prominent impediment, the possibility of therapeutic synergy may perhaps be explored at drastically reduced doses of PALA, combined with other modulating measures.     

Treatment with 5-fluorouracil enhances radiosensitivity of the human pancreatic cancer cell line MiaPaCa-2

Background and Purpose: Several clinical studies have suggested that the combination of radiation therapy and 5-fluorouracil (5-FU) may improve outcome of patients with pancreatic cancer. However, there are few experimental studies supporting this treatment. Aim of the Study: To examine the radiosensitivity of human pancreatic cancer cells and its modulation by 5-FU. Material and Methods: MiaPaCa-2, PANC-1 and NP-18 cells growing as monolayer culture were treated with radiation and 5-FU. In addition, 5-FU was studied administered either pre- or postradiation, both as pulse or continuous exposure. Cell survival was determined by the in vitro clonogenic assay. Results: In MiaPaCa-2 cell line, both radiation and 5-FU alone reduced cell survival. The addition of 5-FU to radiation caused a significant net decrease of cell survival. Pulse exposure of 5-FU decreased survival after 2 Gy and mean inactivation dose by 1.64; continuous exposure decreased survival after 2 Gy and mean inactivation dose by about 2.4. Timing of 5-FU exposure did not modify survival. However, when adjusting for 5-FU killing effect and cell multiplicity, only continuous exposure significantly enhanced radiation cell killing. Conclusion: Both pulse and continuous exposure increase radiation cell killing, but only continuous exposure may radiosensitize MiaPaCa-2 cells.     

氯喹增强LOVO细胞对5-氟尿嘧啶化疗敏感性的作用

目的研究结肠癌细胞LOVO在氯喹(CQ)作用下对化疗药物5-氟尿嘧啶(5-FU)的敏感性变化及机制。方法用MTT法分别检测CQ和5-FU作用24 h和48 h对LOVO细胞的增殖抑制作用。实验分为4组:空白对照组、CQ组、5-FU组和联合作用组。CQ、5-FU及两者联合作用于LOVO细胞后,通过细胞划痕实验和Transwell分别检测细胞迁移和侵袭的变化,采用流式细胞术和TUNEL方法检测细胞凋亡,采用Western blot检测细胞自噬及凋亡相关蛋白的表达情况。结果在不同浓度CQ、5-FU作用下LOVO细胞增殖受到抑制,且呈现剂量依赖性,48 h时CQ和5-FU的IC50分别为11.8μg/ml和2.5μmol/L。与空白对照组相比,CQ、5-FU作用后细胞迁移率和细胞侵袭能力显著降低,且两药联用组的抑制效应更显著。流式细胞术检测显示,CQ组、5-FU组细胞凋亡率分别为(17.85±1.04)%、(17.36±0.96)%,显著高于空白对照组(4.11±0.23)%,两药联用组凋亡率为(25.03±2.27)%,两药联用组与CQ、5-FU单用组相比,细胞凋亡率更高,差异均有统计学意义(P<0.01)。TUNEL检测显示,CQ组、5-FU组细胞凋亡指数显著高于空白对照组,两药联用组与CQ、5-FU单用组相比细胞核凋亡指数更高。Western blot检测显示CQ和5-FU处理后导致P53、Bax、caspase3、caspase9和P62蛋白表达水平显著升高,而Bcl-2、LC3和Beclin-1蛋白表达水平则显著降低,差异均有统计学意义(P<0.01)。结论 CQ能抑制结直肠癌细胞LOVO的增殖、迁移、侵袭和自噬,促进其凋亡,并能增强细胞对化疗药物5-FU的敏感性。     

芦荟大黄素联合氟尿嘧啶对人胃癌细胞增殖及凋亡的影响

目的 探讨芦荟大黄素(AE)与5-氟尿嘧啶(5-FU)体外协同作用及对人胃癌细胞增殖及凋亡的影响.方法 MTT法检测药物对细胞增殖抑制作用;光镜观察细胞生长状态,计数细胞分裂指数;电镜观察药物对细胞超微结构的影响;TUNEL法检测细胞凋亡.结果 AE单独应用对细胞的增殖抑制作用弱,与5-FU合用明显抑制细胞生长,呈剂量依赖性;药物作用48 h后,联合用药组细胞分裂指数显著降低,可见大量细胞皱缩,胞膜界限模糊,核膜消失,核质浓缩及凋亡小体形成;TUNEL法分析显示,联合用药组细胞凋亡率为(69.4±4.1)%,与AE组[(15.7 4±1.9)%]、5-FU组[(33.5±2.8)%]比较差异显著(P<0.05).结论 AE与5-FU联合应用能有效抑制胃癌细胞增殖,促进细胞凋亡,二药具有协同增效作用.     

金雀异黄素联合5-氟尿嘧啶抑制人结肠癌细胞株 SW480增殖的分子机制

目的：初步探讨金雀异黄素提高结肠癌细胞对5-氟尿嘧啶（5-FU）的敏感性,促进结肠癌细胞凋亡的分子机制。方法选择 SW480结肠癌细胞株为实验对象,将金雀异黄素80μmol／L 和5-FU 30μg／mL 单用或联用48 h 作为药物实验组,另设不加药组为对照组。分别采用实时定量 PCR 和Western 印迹法检测各组细胞的生存素、Bcl-2、p21、caspase3和 caspase 9基因和蛋白表达情况。采用比较阈值法（2－ΔΔCT 法）来确定基因的相对表达量。以β-actin 作为内参照,进行半定量分析计算蛋白相对表达量。电泳迁移率改变分析（EMSA）检测各组细胞 NF-κB 的 DNA 结合活性。多组间样本均数比较采用单因素方差分析,组间两样本均数比较采用 LSD 检验。结果联合用药组的 caspase3、caspase9和p21基因表达（1．903±0．122、2．726±0．050、2．541±0．393）与蛋白表达（0．534±0．077、1．161±0．172、0．463±0．016）高于对照组（1．001±0．052、1．000±0．014、1．001±0．037和0．080±0．043、0．248±0．059、0．139±0．021）、金雀异黄素组（1．559±0．038、2．394±0．095、1．686±0．061和0．335±0．052、0．478±0．059、0．304±0．018）和5-FU 组（1．198±0．063、1．051±0．043、1．399±0．055和0．194±0．015、0．337±0．036、0．231±0．011）,联合用药组的生存素基因与蛋白表达（0．165±0．018和0．216±0．014）低于对照组、金雀异黄素组和5-氟尿嘧啶单组（1．001±0．033、0．775±0．044、0．395±0．030和0．594±0．079、0．375±0．014、0．295±0．014）,差异均有统计学意义（基因表达,F ＝802．865、52．760、39．992、187．288、37．435;蛋白表达,F ＝10．466、44．483、19．490、200．011、45．238;P 均＜0．01）, caspase3、p21在两单药组的表达与对照组相比,差异也有统计学意义（LSD 检验,P ＜0．05）。EMSA 检测显示,与对照组（1067．97±36．01）和5-FU 组（718．83±23．18）相比,金雀异黄素组（461．64±15．41）和联合用药组（585．28±7．82）的 NF-κB 蛋白 DNA 结合活性均有明显降低（LSD 检验,P ＜0．05）。结论金雀异黄素联合5-FU 协同促进结肠癌细胞凋亡,可能通过金雀异黄素抑制 NF-κB 的 DNA 结合活性,进一步上调 caspase3、caspase9、p21以及下调生存素起作用,提示金雀异黄素可能为结肠癌化学治疗提供辅助。     

Antiproliferative effects of 5-fluorouracil and interferon-alpha in combination on a hepatocellular carcinoma cell line in vitro and in vivo

Background and Aim: We investigated the antiproliferative effects of interferon‐alpha (IFN‐α) and 5‐fluorouracil (5‐FU) in combination on a hepatocellular carcinoma (HCC) cell line. Method: In the in vitro study, IFN‐α and/or 5‐FU was added to the culture of the poorly differentiated‐type HCC cell line, HAK‐1B, and their antiproliferative effects and additional or synergic effects in combination treatment were examined. In the in vivo study, HAK‐1B cells were transplanted into nude mice and the changes in tumor volume and weight, apoptosis, BrdU and cyclin A positive cells, and artery‐like blood vessels were investigated. Expressions of angiogenesis factors and IFN‐α receptor (IFNAR‐2) were examined in the developed tumors. Results: In vitro growth of HAK‐1B cells was suppressed dose‐dependently to 5‐FU, but the addition of IFN‐α did not induce additional or synergic effects. In vivo growth in terms of tumor diameter and weight was suppressed at most in the IFN‐α + 5‐FU (combination) group, that is, the tumor volume became 29.3% and the tumor weight became 54.7% of the control. In the combination group, numbers of BrdU‐positive S‐phase cells and cyclin A positive cells increased together with the increase in apoptotic cells, but there was no significant relation between the tumor shrinkage effects and angiogenesis factors or artery‐like blood vessels. In the combination group, INFAR‐2 decreased significantly in comparison to the other groups. Conclusion: The synergic growth‐suppression effects in the current in vivo study using the combination treatment are attributable to the enhanced induction of S‐phase arrest and of apoptosis.     

Use of 5-fluorouracil and survival in patients with microsatellite-unstable colorectal cancer

BACKGROUND & AIMS: 5-Fluorouracil improves mortality in stage III colorectal cancer patients. In vitro studies suggest that microsatellite instability influences cell survival after 5-fluorouracil treatment. We investigated the survival influence of 5-fluorouracil in patients with microsatellite instability-high tumors. METHODS: We collected data and tumors on 204 consecutive stage II and III colorectal cancer patients from registries at the University of California and Veterans Administration hospitals in San Diego, California, from 1982 to 1999. Archival DNA was extracted, and microsatellite instability was assessed by National Cancer Institute-recommended markers. Cox proportional hazard modeling was used to determine survival associations for microsatellite instability and 5-fluorouracil treatment status. RESULTS: We identified 36 microsatellite instability-high (17.6%) and 168 non-microsatellite instability-high tumors (82.4%). Microsatellite instability-high tumors were significantly associated with proximal colon location, presence of mucin, and surrounding lymphoid reaction. Univariate and multivariate analyses showed no survival difference between microsatellite instability-high and non-microsatellite instability-high groups (hazard ratio, 1.04; P = 0.88). Dichotomized by use of 5-fluorouracil, there was increased risk of death in patients who received no adjuvant chemotherapy (hazard ratio, 2.02; P = 0.02). However, the benefit of 5-fluorouracil was different between microsatellite instability-high and non-microsatellite instability-high groups. Patients with non-microsatellite instability-high tumors who received 5-fluorouracil had better survival compared with patients who were not treated (P < 0.05). Conversely, patients with microsatellite instability-high tumors who were treated with 5-fluorouracil had no survival difference compared with patients without treatment (P = 0.52). CONCLUSIONS: There is improved survival in patients with non-microsatellite instability-high tumors after 5-fluorouracil-based chemotherapy that does not extend to patients with microsatellite instability-high tumors. The microsatellite instability status of a patient's colorectal cancer may indicate differences in 5-fluorouracil-based chemosensitivity; this is consistent with in vitro studies.     

Anti-mitogenic and apoptotic effects of 5-HT1B receptor antagonist on HT29 colorectal cancer cell line

Purpose  

There is lack of evidence about impact of 5-HT receptors on colorectal cancers. The current study was designed to investigate the role of serotonin and its receptors in colorectal cancer cell line and tissues.     

Gefitinib as a first line of therapy in non-small cell lung cancer with brain metastases

Two Japanese women were diagnosed as having well-differentiated adenocarcinoma of the lung with brain metastases. Since it was considered they could not tolerate conventional chemotherapy, we administered gefitinib without any previous systemic therapy. In both patients gefitinib acted dramatically on all the lesions including the brain metastases, resulting in a marked decrease of the elevated CEA levels, and improvement of their quality of life. Retrospective evaluation of epidermal growth factor receptor expression levels by immunohistochemistry revealed positive results in both cases. Though gefitinib has been recommended for patients previously treated with chemotherapy, it should be considered feasible as a first line therapy.     

草酸铂及羟基喜树碱联合亚叶酸钙和氟尿嘧啶治疗进展期结直肠癌的比较

目的：比较草酸铂及羟基喜树碱联合亚叶酸钙、氟尿嘧啶治疗进展期结直肠癌的近期疗效、不良反应和生存期．方法：经病理证实的进展期结直肠癌病例60例,40例(OLF组)采用草酸铂联合亚叶酸钙和氟尿嘧啶方案化疗92个周期,20例(HLF组)采用羟基喜树碱联合亚叶酸钙和氟尿嘧啶方案化疗40个周期,观察化疗不良反应,2-3周期后评价疗效,随访观察无疾病进展生存期、总生存期,统计1a生存率．结果：OLF和HLF组近期有效率分别为30．0%(12/40)和25．0%(5/20),无统计学差异(χ~2 =0．531,P=0．811)；中位无疾病进展生存期分别为6．4和7．3 mo,无统计学差异(u=1．5088,P＞0．05)；中位总生存期分别为10．2和10．8 mo,无统计学差异(u=0．3487,P＞0．05)；1 a生存率分别为34．09%和38．55%,无统计学差异(u= 0．3275,P＞0．05)．两组间Ⅲ,Ⅳ度不良反应均以骨髓抑制和消化道反应为主,其中腹泻发生率HLF组高于OLF组,有统计学差异(χ~2=7．876,P=0．044)．结论：OLF和HLF两方案治疗进展期结直肠癌疗效相似,前者外周神经毒性较常见,后者腹泻更常见．     

Hypofractionated radiotherapy with concurrent 5-fluorouracil radiosensitisation for recurrent or locally advanced colorectal cancer A phase II study

In a pilot study we treated 19 patients suffering from recurrent or locally advanced inoperable colorectal cancer, with concurrent hypofractionated radiotherapy (4 – 5 Gy/fraction, 2 fractions per week) and 5-Fluorouracil bolus, 1 hour before RT at doses of 300 mg/m². For 16 patients treated with radical intent the Normalised Total Dose for α/β = 10 Gy ranged between 56 – 74 Gy (median 62 Gy). The schedule used was very well tolerated. Moderate grade II haematological toxicity was observed in 11% of cases and diarrhoea grade II/III resulting in 2 – 4 weeks treatment delay was observed in 26% of cases. One case with bowel perforation and one with painful subcutaneous fibrosis was observed during 12 – 27 months of follow-up. Out of 16 patients treated with radical intent 4 (25%) showed complete response and the overall response rate was 56% (9/16). The one year symptom free survival was obtained in 11/16 (69%) radically treated patients. It is concluded that hypofractionated radiochemotherapy with 5-Fluorouracil for recurrent or locally advanced colorectal cancer is an effective regimen and has acceptable acute and late toxicity. Further investigation is required.

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Résumé. Dans un projet pilote, nous avons traité 19 patients souffrant d'un cancer colo-rectal récidivant ou localement avancé inopérable par l'administration simultanée d'une radiothérapie hypofractionnée (4 – 5 Gy/fraction à raison de 2 fractions par semaine) et bolus de 5-Fluorouracil à la dose de 300 mg/m² une heure avant la radiothérapie. Chez 16 patients traités dans un but radical, la dose totale normalisée a été de 56 – 74 Gy (moyenne 62 Gy). Le schéma utilisé a été particulièrement bien toléré. Une toxicité hématologique modérée de grade II a été observée chez 11% des patients et une diarrhée de grade II/III entra?nant un délai dans le traitement de 2 à 4 semaines a été observée chez 26% des patients. Une perforation intestinale et une fibrose sous-cutanée douloureuse ont été observées au cours de la période postopératoire de 12 à 27 mois. Quatre des 16 patients traités à des fins curatives (25%) ont montré une résponse complète et une réponse globale a été observée chez 56% des patients (9/16). Une survie sans sympt?mes à un an a été obtenue chez 11 des 16 patients (69%). On conclut que l'assocation de 5-Fluorouracil et d'une radiothérapie hypofractionnée constitue un schéma thérapeutique effectif en cas de cancer colorectal récidivant ou localement avancé avec une toxicité aigu? et tardive acceptable. Des investigations complémentaires sont nécessaires.

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Accepted: 20 June 1996     

脂质体包裹EGFR反义寡脱氧核苷酸抑制人大肠癌细胞的增殖

目的研究用表皮生长因子受体（ＥＧＦＲ）反义寡脱氧核苷酸（ＡＳＯＤＮ）抑制人大肠癌细胞株ＨＲ８３４８的生长增殖，探讨大肠癌治疗的新途径．方法人工合成的ＥＧＦＲ反义ＯＤＮ及无关对照ＯＤＮ经脂质体包裹后作用ＨＲ８３４８细胞，采用ＭＴＴ法，［３Ｈ］ＴｄＲ掺入试验，细胞周期分析，裸鼠体内接种等方法测定细胞体内外增殖的变化．结果经ＥＧＦＲ反义ＯＤＮ处理的ＨＲ８３４８细胞与对照组ＨＲ８３４８细胞相比，体外增殖速度减慢（细胞增殖抑制率达７１％）、ＤＮＡ合成受抑（细胞增殖指数为０３６１ｖｓ０７８４，Ｓ期细胞数为３４８％ｖｓ５９９％）、裸鼠体内成瘤率下降（２５％ｖｓ１００％）．结论ＥＧＦＲ反义ＯＤＮ可有效抑制大肠癌细胞的体内外生长增殖，可用于实验性肿瘤基因治疗研究     

Randomized trial comparing weekly bolus 5-fluorouracil plus leucovorin versus monthly 5-day 5-fluorouracil plus leucovorin in metastatic colorectal cancer

BACKGROUND/AIMS: The purpose of this study was to compare the efficacy and toxicity profiles of weekly intravenous (i.v.) bolus injection of 5-fluorouracil plus low-dose leucovorin with the Mayo Clinics' monthly 5-day schedule of 5-fluorouracil and leucovorin in the treatment of metastatic colorectal cancer. METHODOLOGY: A total of 96 patients with previously untreated metastatic colorectal cancer were randomized to receive either a weekly i.v. bolus injection of 5-fluorouracil 400 mg/m2 plus leucovorin 20 mg/m2 (weekly arm), or i.v. bolus injection of 5-fluorouracil 425 mg/m2 plus leucovorin 20 mg/m2 for 5 consecutive days every 4 or 5 weeks (monthly arm). Therapy was continued until disease progression or unacceptable toxicity appeared. In the presence of disease progression, the study regimen was stopped and second-line treatment was instituted after the patient was discontinued from this study. RESULTS: There was no significant difference of response rates between both regimens. The response rate were 14.3% in the weekly arm (2 CR and 5 PR, 95% CI: 2.6-25.2%) and 10.6% in the monthly arm (1 CR and 4 PR; 95% CI: 6.5-32.3%), respectively (P = 0.8957). The survival times were also similar between the two (P = 0.4207, log-rank test). The median survival were 15.8 months in the monthly arm and 18.4 months in the weekly arm. Hematologic toxicity was minimal in both arms. However, the monthly arm produced a higher toxicity in severe (grade 3-4) diarrhea (14.9% vs. 2%; P = 0.029) and stomatitis (8.5% vs. 0; P = 0.054). CONCLUSIONS: Weekly bolus injection of 5-fluorouracil and low-dose leucovorin achieved a similar response rate and survival as compared with the Mayo Clinics' monthly 5-day schedule, but severe toxicity was less commonly seen using the weekly regimen. As current chemotherapeutic treatment for metastatic colorectal cancer is largely palliative rather than curative, the weekly bolus regimen may be a more favorable approach in managing metastatic colorectal cancer.     

Should capecitabine replace 5-fluorouracil in the first-line treatment of metastatic colorectal cancer?

Fluoropyrimidines play a central role in the first-line treatment of metastatic colorectal cancer. Our aim was to review whether capecitabine was a safer, non-inferior, economically superior and more convenient alternative to 5-fluorouracil. Capecitabine has previously been compared to 5-fluorouracil-either as a monotherapy or in combination with oxaliplatin, irinotecan, or biological drugs-and has been found to have comparable efficacy and safety profiles. Furthermore, pharmacoeconomic data and patients’ preferences for oral chemotherapy further favor capecitabine. Therefore, capecitabine appears to be an effective and safe alternative to fluorouracil in the first-line treatment of metastatic colorectal cancer.     

Pilot trial of prolonged continuous-infusion 5-fluorouracil and weekly cisplatin in advanced colorectal cancer

Thirty-five patients with previously untreated, advanced, measurable metastatic colorectal carcinoma were treated with a 12-week course of continuous 5-fluorouracil (5-FU) and weekly cisplatin. Twenty of 32 evaluable patients responded (five complete and 15 partial responses), for an overall response rate of 63% (90% confidence limits, 43%-75%). Toxicity was generally mild and reversible and included mucositis (40%), painful erythema of the palmar and plantar skin (30%), diarrhea (21%), nausea and vomiting (15%), and leukopenia (6%). One patient died of sepsis secondary to mucositis and myelosuppression. This program is a well-tolerated outpatient regimen for metastatic colorectal carcinoma. The response rate is higher than expected for 5-FU and cisplatin and suggests clinical therapeutic synergism at this dose rate and schedule.