Establishment of organoid models based on a nested array chip for fast and reproducible drug testing in colorectal cancer therapy

The conventional microwell-based platform for construction of organoid models exhibits limitations in precision oncology applications because of low-speed growth and high variability. Here, we established organoid models on a nested array chip for fast and reproducible drug testing using 50% matrigel. First, we constructed mouse small intestinal and colonic organoid models. Compared with the conventional microwell-based platform, the mouse organoids on the chip showed accelerated growth and improved reproducibility due to the nested design of the chip. The design of the chip provides miniaturized and uniform shaping of the matrigel that allows the organoid to grow in a concentrated and controlled manner. Next, a patient-derived organoid(PDO) model from colorectal cancer tissues was successfully generated and characterized on the chip. Finally, the PDO models on the chip, from three patients, were implemented for high-throughput drug screening using nine treatment regimens. The drug sensitivity testing on the PDO models showed good quality control with a coefficient of variation under 10% and a Z’ factor of more than 0.7. More importantly, the drug responses on the chip recapitulate the heterogeneous response of individual patients, as well as showing a potential correlation with clinical outcomes. Therefore,the organoid model coupled with the nested array chip platform provides a fast and reproducible means for predicting drug responses to accelerate precise oncology.

Trans-arterial chemoperfusion for the treatment of liver metastases of breast cancer and colorectal cancer: Clinical results in palliative care patients

AIM To evaluate the clinical value and efficiency of transarterial chemoperfusion(TACP) in patients with liver metastases from breast cancer(BC) and colorectal cancer(CRC).METHODS We treated 36 patients with liver metastases of BC(n = 19, 19 females) and CRC(n = 17; 8 females, 9 males) with repeated TACP. The treatment interval was 4 wk. TACP was performed with gemcitabine(1000 mg/m2) and mitomycin(10 mg/m2), administered within 1 h after positioning the catheter tip in the hepatic artery. Before treatment, the size, location, tumour volume, vascularization and number of liver tumours were evaluated using magnetic resonance imaging(MRI). Tumour response was evaluated according to the Response Evaluation Criteria in Solid Tumors guidelines.RESULTS TACP using gemcitabine and mitomycin for metastases from CRC and BC was performed without any serious side effects. The follow-up MRI showed a therapeutic response in 84.2% of the BC patients-stable disease 47.4% and partial response 36.8%. A progression was seen in 15.8%.CRC patients showed a therapeutic response in 52.9% of cases. A progression of the disease was documented in 47.1% of the patients with CRC. These data show that TACP in patients with liver metastases of BC leads to a significantly better therapeutic response compared with CRC patients(P = 0.042). The median survival time was 13.2 mo for the BC patients, which is significantly longer than for CRC patients at 9.3 mo(P = 0.001).CONCLUSION TACP for liver metastases of BC appears to be a safe and effective palliative treatment with improved outcomes in comparison to patients with CRC.

Liver-first approach of colorectal cancer with synchronous hepatic metastases: A reverse strategy

Recently, there has been a change in the strategy of how synchronous colorectal hepatic metastases are attributed to the development of more valuable protocols of chemotherapy and radiotherapy for neoadjuvant treatment of colorectal neoplasms and their hepatic metastases. There is a consensus that patients with synchronous colorectal hepatic metastases have lower survival than those with metachronous colorectal hepatic metastases. Currently, controversy remains concerning the best approach is sequence in a patient with colorectal cancer and synchronous hepatic metastases resection. To obtain a better patient selection, the authors have suggested the initial realization of systemic chemotherapy in the circumstance of patients with colorectal tumor stage Ⅳ, since these patients have a systemic disease. The rationale behind this liver-first strategy is initially the control of synchronous hepatic metastases of colorectal carcinoma, which can optimize a potentially curative hepatic resection and longstanding survival. The liver-first strategy procedure is indicated for patients with colorectal hepatic metastases who require downstaging therapy to make a curative liver resection possible. Thus, the liver-first strategy is considered an option in cases of rectal carcinoma in the early stage and with limited or advanced synchronous colorectal hepatic metastases or in case of patients with asymptomatic colorectal carcinoma, but with extensive liver metastases. Patients undergoing systemic chemotherapy and with progression of neoplastic disease should not undergo hepatic resection, because it does not change the prognosis and may even make it worse. To date, there have been no randomized controlled trials on surgical approach of colorectal synchronous hepatic metastases, despite the relatively high number of available manuscripts on this subject. All of these published studies are observational, usually retrospective, and often non-comparative. The patient selection criteria for the liver-first strategy should be individualized, and the approach of these patients should be performed by a multidisciplinary team so its benefits will be fully realized.

Effect of a nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester on invasion of human colorectal cancer cell line SL-174T

AIML To investigate the effect and mechanism of action of the nitric oxide synthase （NOS） inhibitor NG-nitro-L-arginine methyl ester （L-NAME） on invasion and metastasis of human colorectal cancer cell line SL-174T. METHODS： Human colorectal cancer cel4 line SL-174T was cultured and treated separately with four different dosages of L-NAME for 72 h, Nitric oxide （NO） production was measured with Griess reagent, The effect of L-NAME on invasion and migration of SL-174T cells were evaluated by using Transwell chambers attached with polycarbonate filters and reconstituted basement membrane （Matrigel）, RT-PCR was performed to determine the mRNA levels of matrix metalloproteinase-2 （MMP-2） and tissue inhibitor metalloproteinase-2 （TIMP-2）,RESULTS： L-NAME could significantly inhibit NO production of SL-174T in a dose-dependent manner. After being treated for 72 h with 0.2, 0.4, 0.8, and 1.0 mmol/L L- NAME, respectively, the ability of the L-NAME treated SL- 174T cells to invade the reconstituted basement membrane decreased significantly （t = 8.056, P〈0.05; t= 14.467, P〈0.01; t= 27.785, P〈0.01; and t= 29.405, P〈0.01, respectively） and the inhibition rates were 10.29%, 19.62%, 34.08%, and 42.23%, respectively. Moreover, L-NAME could inhibit migration of SL-174T cells, and the inhibition rates were 20.76%, 24.95%, 39.43%, and 46. 85% for L-NAME at 0.2, 0.4, 0.8, and 1.0 mmol/L, respectively （t = 15.116, P〈0.01）. In addition, after treatment with L-NAME, expression of MMP-2 mRNA was significantly decreased （t = 71.238, P〈0.01） and that of TIMP-2 mRNA was markedly increased （t = -13.020, P〈0.01）. CONCLUSION： L-NAME exerts anti-invasive and anti- metastatic effects on SL-174T cell line via downregulating MNP-2 mRNA expression and upregulating TIMP-2 mRNA expression.

Autophagy-related proteins Beclin-1 and LC3 predict cetuximab efficacy in advanced colorectal cancer

AIM:To investigate the utility of Beclin-1 and LC3,two autophagy-related proteins,in predicting the cetuximab efficacy in advanced colorectal cancer(ACRC) . METHODS:The data of 85 patients with ACRC treated at the Sun Yat-sen University Cancer Center from March 1,2005 to December 31,2008 were studied,including 45 cases treated with cetuximab-containing chemotherapy and 40 cases treated with non-cetuximab-containing chemotherapy.Beclin-1 and LC3 expression was evaluated by immunohistochemistry,and KRAS status was evaluated by polymerase chain reaction. RESULTS:Beclin-1 and LC3 expression in ACRC wassignificantly correlated(r=0.44,P<0.01);however,LC3 was more highly expressed in cancerous tissues than in normal tissues(Z=-2.63,P<0.01) .In the cetuximab-containing chemotherapy group,patients with low LC3 expression had higher objective response rates(ORRs) than those with high LC3 expression(52.9%vs 17.9%,P=0.01) ,and patients with low Beclin-1 expression had a longer median progressionfree survival(PFS) than their counterparts with higher Beclin-1 expression(9.0 mo vs 3.0 mo,P=0.01) . However,neither of these predictive relationships was detected in the group treated with non-cetuximabcontaining chemotherapy.Patients with wild-type KRAS had higher ORRs(42.3%vs 9.1%,P=0.049) and disease control rates(DCRs)(73.1%vs 36.4%,P= 0.035) ,and longer median PFS(5.5 mo vs 2.5 mo,P= 0.02) than those with mutant KRAS in the cetuximabcontaining chemotherapy group.Neither Beclin-1(P= 0.52) nor LC3(P=0.32) expression was significantly correlated with KRAS status. CONCLUSION:Patients with low Beclin-1 expression had a longer PFS than those with high Beclin-1 expression,and patients with low LC3 expression had a higher ORR in ACRC patients treated with cetuximab-containing chemotherapy.

Glycogen Synthase Kinase 3β Inhibitor (2'Z,3'E)-6-Bromo-indirubin-3'-Oxime Enhances Drug Resistance to 5-Fluorouracil Chemotherapy in Colon Cancer Cells

Objective： To explore the effects and mechanism of glycogen synthase kinase 3β （GSK-313） inhibitor （2＇Z,3＇E）-6-bromo-indirubin-3＇-oxime （BIO） on drug resistance in colon cancer cells. Methods： The colon cancer SW480 and SW620 cells were treated with BIO, 5-fluorouracil （5-FU） and BIO/5-FU, separately. Cell cycle distribution, apoptosis level and efflux ability of rhodamine 123 （Rh123） were detected by flow cytometry. The protein expressions of P-glycoprotein （P-gp）, multidrug resistance protein 2 （MRP2）, thymidylate synthase （TS）, β-catenin, E2F-1 and βcl-2 were detected by Western blot. β-catenin and P-gp were stained with double immunofluorescence and observed under a confocal microscope. Results： BIO up-regulated β-catenin, P-gp, MRP2 and TS, enhanced the efflux ability of Rh123, decreased Bcl-2 protein and gave the opposite effect to E2F-1 protein in SW480 and SW620 ceils. Furthermore, BIO significantly inhibited cell apoptosis, increased S and G2/M phase cells, and reduced the cell apoptosis induced by 5-FU in SW480 cells, whereas the effects were slight or not obvious in SW620 cells. Conclusion： GSK-3β was involved in drug resistance regulation, and activation of β-catenin and inhibition of E2F-1 may be the most responsible for the enhancement of 5-FU chemotherapy resistance induced by GSK-β inhibitor β10 in colon cancer.

Feasibility and Efficacy Study of Biweekly Irinotecan Combined with Oral Tegafur/Uracil in Advanced Colorectal Cancer

Background: We evaluated the feasibility and efficacy of irinotecan (CPT-11) plus tegafur/uracil (UFT) combination chemotherapy in patients with advanced colorectal cancer. Patients and Methods: PK parameters were concurrently measured to confirm the presence of drug interactions in this treatment schedule. CPT-11 was administered intravenously at the dose of 150 mg/m2 on days 1, 15. UFT was administered at the dose of 375 mg/m2/day (B.I.D.) on days 3 - 7, 10 - 14, 17 - 21, 24 - 28 repeated every 5 weeks. Results: 31 patients were enrolled. PK parameters for CPT-11, FT, 5-FU and uracil are available from 5 patients. The overall response rate was 16.1%. The median time to treatment discontinuation was 3.9 months. There was no significant difference in PK parameters of CPT-11 between day 1 and day 15 and of UFT between day 3 and day 10. Conclusion: CPT-11 plus UFT combination chemotherapy exhibited a tolerable toxicity profile with acceptable efficacy. Pharmacokinetic analysis showed that there were no drug interactions in this treatment schedule.

下调miR-208a通过靶向SFRP1介导Wnt信号通路对结直肠癌细胞5-FU耐药的改善作用

目的:探讨下调微小RNA-208a(miR-208a)对结直肠癌细胞5-氟尿嘧啶(5-FU)耐药的影响,阐明其相关分子机制。方法:采用实时荧光定量PCR(RT-qPCR)法检测结直肠癌5-FU耐药细胞株HT-29/5-FU及其亲本HT-29细胞中miR-208a和分泌型卷曲相关蛋白1(SFRP1)mRNA表达水平。以HT-29/5-FU细胞为研究对象,将miR-208a抑制物(miR-208a inhibitor)质粒及其阴性对照质粒(inbibitor-NC)和SFRP1小干扰质粒(si-SFRP1)及其阴性对照质粒(si-NC)分别或同时转染至HT-29/5-FU细胞中,联合5-FU处理,将细胞分为空白组、inhibitor-NC组、miR-208a inhibitor组、miR-208a inhibitor+si-NC组和miR-208a inhibitor+si-SFRP1组。MTT法检测各组细胞增殖活性并计算耐药指数,AnnexinⅤ-FITC/PI双染法结合流式细胞术检测不同浓度5-FU作用后各组细胞凋亡率,Western blotting法检测各组细胞中SFRP1、β-连环蛋白(β-catenin)、P-糖蛋白(P-gp)和ATP结合盒B亚家族成员1转运蛋白(ABCB1)蛋白表达水平。双荧光素酶报告基因实验验证miR-208a与SFRP1的靶向关系。结果:与HT-29细胞比较,HT-29/5-FU细胞中miR-208a表达水平升高(P<0.05),SFRP1 mRNA表达水平降低(P<0.05)。与inhibitor-NC组比较,miR-208a inhibitor组细胞增殖活性降低(P<0.05),耐药指数降低,细胞凋亡率升高(P<0.05),细胞中β-catenin、P-gp和ABCB1蛋白表达水平降低(P<0.05)。双荧光素酶报告基因实验提示SFRP1是miR-208a靶基因,且miR-208a可负向调控SFRP1的表达。与miR-208a inhibitor+si-NC组比较,miR-208a inhibitor+si-SFRP1组细胞增殖活性升高(P<0.05),耐药指数升高,细胞凋亡率降低(P<0.05),细胞中β-catenin、P-gp和ABCB1蛋白表达水平升高(P<0.05)。结论:下调miR-208a可通过靶向上调SFRP1表达抑制Wnt信号通路的转导,进而改善HT-29/5-FU细胞对5-FU的耐药。

Therapeutic targets of Traditional Chinese Medicine for colorectal cancer

OBJECTIVE: To explore the role of Traditional Chinese Medicine(TCM) in the prevention and treatment of colorectal cancer and identify possible therapeutic targets of TCM to provide clues for the use of TCM for colorectal cancer prevention and treatment in the clinic and to find novel directions for new drug discovery for colorectal cancer.METHODS: We used Pub Med and Google to search for and collect scientific publications for a full evaluation of current evidence in the literature indicating the potential role of Chinese herbal medicines and their respective ingredients as effective candidatesforcolorectalcancerpreventionandtreatment.RESULTS: We extracted a detailed description of potential therapeutic Chinese herbal medicines and their constituent ingredients that target different mechanisms in colorectal cancer such as gene mutation, dysregulation of signaling pathways, metabolism disorders, and the inflammatory microenvironment, including both conventional and non-conventional approaches.CONCLUSION: TCM may be a promising complementary and alternative therapy for the treatment of colorectal cancer.

Targeting KRAS G12C mutations in colorectal cancer

With the advent of Kirsten rat sarcoma viral oncogene homologue G12C(KRAS G12C)inhibitors,RAS is no longer considered undruggable.For the suppression of RAS,new therapeutic approaches have been suggested.However,current clinical studies have indicated therapeutic resistance after short-lived tumour suppression.According to preclinical studies,this might be associated with acquired genetic alterations,reactivation of downstream pathways,and stimulation for upstream signalling.In this review,we aimed to summarize current approaches for combination therapy to alleviate resistance to KRAS G12C inhibitors in colorectal cancer with a focus on the mechanisms of therapeutic resistance.We also analysed the relationship between various mechanisms and therapeutic resistance.

Effect of mild moxibustion on cancer-related fatigue,serum ghrelin and adiponectin in patients undergoing chemotherapy after colorectal cancer surgery

Objective To observe the effect of mild moxibustion on cancer-related fatigue,serum ghrelin and adiponectin in patients undergoing chemotherapy after colorectal cancer surgery.Methods Seventy patients were divided into a control group and an observation group according to the random number table method,with 35 cases in each group.Patients in both groups were treated with conventional FOLFOX4 chemotherapy regimen.Patients in the control group also received routine treatments such as symptomatic treatment of complications,health education,nutritional support,and exercise intervention.Patients in the observation group received mild moxibustion on the basis of interventions used in the control group.The Piper fatigue scale-revision(PFS-R),Karnofsky performance status(KPS)and spleen and kidney yang deficiency syndrome were scored,and serum ghrelin and adiponectin levels were measured before and after treatment.Results After treatment,total score of PFS-R in the control group did not change significantly(P>0.05),while in the observation group it was significantly reduced(P<0.05)and lower than that in the control group(P<0.05);KPS scores in both groups were increased(both P<0.05),and it was significantly higher in the observation group than in the control group(P<0.05);total score of spleen and kidney yang deficiency syndrome in the control group did not change significantly(P>0.05),while it was significantly reduced in the observation group(P<0.05)and lower than that in the control group(P<0.05);serum ghrelin and adiponectin levels in the control group had no significant changes(both P>0.05),but the levels were significantly increased in the observation group(both P<0.05)and higher than those in the control group(both P<0.05).Conclusion Based on the routine treatments,mild moxibustion can relieve fatigue,improve quality of life,and improve the symptoms of spleen and kidney yang deficiency in patients undergoing chemotherapy after colorectal cancer surgery,which may be related to the regulation of serum ghrelin and adiponectin levels.

奥沙利铂单药或与氟尿嘧啶-甲酰四氢叶酸联合应用治疗晚期大肠癌Ⅱ期临床试用报告

目的：观察奥沙利铂治疗晚期大肠癌的临床疗效和安全性。方法：根据ＧＣＰ指导原则开展多中心Ⅱ期随机临床研究：Ａ组奥沙利铂单药１３０ｍｇ／ｍ２静脉滴注２ｈ，每３周一次；Ｂ组奥沙利铂１３０ｍｇ／ｍ２静脉滴注２ｈＤ１，加甲酰四氢叶酸钙（ＣＦ）２００ｍｇ／ｍ２＋５氟尿嘧啶３００ｍｇ／ｍ２静脉滴注４ｈＤ１到Ｄ５，每３周一次。治疗３周期后评定疗效，有效病例在４周后确认疗效。结果：共收入１１４例，在可统计近期疗效的１００例中，ＣＲ１例，ＰＲ２６例，ＳＤ３２例，ＰＤ４１例，总有效率２７０％。Ａ组３６例单药治疗有效率为１３９％；Ｂ组６４例有效率为３４４％。不良反应主要为恶心、呕吐、贫血和感觉神经毒性。对白细胞和血小板影响较小。结论：经考核奥沙利铂对大肠癌疗效和不良反应与欧洲的结果相近，和氟尿嘧啶合用疗效突出；多数病人耐受良好，是一治疗大肠癌有希望的新药。进一步考核，奥沙利铂和氟尿嘧啶联合有望成为治疗晚期大肠癌的首选方法。

人增殖抑制基因(HSG)对肿瘤细胞系化疗敏感性的作用

目的:将人增殖抑制基因 (hHSG)用电子穿孔的转基因方式转染到体外培养的人肿瘤细胞系中,观察hHSG基因对内源性hHSG表达水平不同的肿瘤细胞系的化疗敏感性的影响。方法:首先用免疫组化方法检测不同组织来源的肿瘤细胞系中hHSG的表达水平,然后选择内源性hHSG表达水平较低的肺腺癌 (A549 )和内源性hHSG表达水平较高的宫颈癌(HeLaS3)细胞系,用电穿孔方法转染含有hHSG的真核表达载体 (pEGFP hHSG) 24h后,加入放线菌酮(CHX),采用细胞计数、MTT法观察hHSG对肿瘤细胞增殖抑制作用及对CHX的化疗敏感性的影响。结果:hHSG在不同组织来源的肿瘤细胞系都有不同程度的表达, pEGFP hHSG转染到两种内源性hHSG表达水平不同的肿瘤细胞系后,这两种肿瘤细胞的生长增殖都明显受到抑制,同时外源性的hHSG也增强了这些肿瘤细胞系对CHX的敏感性。结论:外源性hHSG可不依赖其内源性表达水平而抑制肿瘤细胞的增殖,与CHX并用可增强肿瘤细胞对CHX的敏感性,具有协同作用。

吉西他滨区域性动脉灌注联合全身化疗治疗晚期胰腺癌

目的评价吉西他滨区域性动脉灌注联合全身化疗对晚期胰腺癌的治疗效果。方法对13例经手术病理或临床证实的晚期胰腺癌患者采用5-FU+MMC行静脉化疗,以介入方法用吉西他滨作区域性动脉灌注化疗。结果13例可评价疗效者中部分缓解(PR)4例,病情进展(SD)6例,PD3例;临床受益反应评价有效率为76.9%;疼痛缓解率75.0%。中位生存时间为6.3个月,所有患者未出现严重毒副反应。结论吉西他滨区域性动脉灌注联合全身化疗可缓解晚期胰腺癌患者癌性疼痛,改善患者一般状态,提高生存质量,延长生存期,且患者耐受良好。

晚期乳腺癌术前局部灌注化疗的疗效观察

目的 探讨术前动脉灌注化疗对晚期乳腺癌临床治疗效果及病理特征的影响。方法 回顾性分析 5 2例晚期乳腺癌患者的临床资料 ,其中 2 2例术前行动脉内灌注化疗 (治疗组 ) ,3 0例术前未行动脉内灌注化疗 (对照组 )。结果 治疗组灌注化疗后 ,症状减轻 ,肿瘤缩小 ,有效率 (CR +PR )为 86.4%。术后病理检查均发现癌细胞核固缩、碎裂 ,胞浆凝固、坏死 ;细胞间质水肿、炎性细胞浸润、纤维组织增生 ;血管出现内膜增生 ,血栓形成。对照组癌细胞改变不明显。随访时间 2～ 7年。局部复发率治疗组 13 .6% ,对照组 3 3 .3 % (P <0 .0 1)。治疗组 5年生存率 5 9.1% ,对照组 2 6.7%(P <0 .0 5 )。结论 晚期乳腺癌术前动脉灌注化疗可以缩小肿瘤 ,降低肿瘤分期 ,改变癌细胞的组织学形态 ,提高生存率。

5-氟尿嘧啶缓释剂瘤内注射治疗胰腺癌的实验研究和临床研究

目的 观察5 -氟尿嘧啶( 5 FU)缓释剂对荷胰腺癌裸鼠肿瘤细胞及胰腺癌患者血清肿瘤标记物和细胞免疫的影响。方法 ( 1 ) 5 FU缓释剂的体外释放实验和体外抑瘤实验:测定浸出液药物的浓度,计算释放量;检测其浸出液对人胰腺癌细胞株PC3的抑制作用。( 2 )将荷胰腺癌细胞株PC3裸鼠6 0只,随机分成静脉对照组(A组)、5 FU静注组(B组)、基质植入组(C组)、大剂量5 FU缓释剂植入组(D组)和小剂量5 FU缓释剂植入组(E组)。治疗前及治疗后1 4 d测肿瘤大小。治疗2周后观察肿瘤组织学变化。免疫组化法测定bcl 2和Bax的蛋白表达水平;TUNEL法检测凋亡指数(AI)。( 3 )手术探查不能切除之胰腺癌6 9例随机分成3组。将5 FU缓释剂瘤内植入治疗组(治疗组)、术后行5 FU静脉化疗组(化疗组)和对照组。分别于术前1d和术后第1 4天采血,测定各组血清中NK细胞,T细胞亚群和CEA,CA5 0,CA1 9 9,CA1 2 5,CA2 4 2血清肿瘤标记物水平。结果 ( 1 ) 5 mg5 FU缓释剂第1天释放量最大,为0. 8 5 mg,第3天为0. 4 5 mg,其后在0. 2 5 mg水平维持稳定的缓慢释放;释放时间长达1 4 d以上。( 2 ) 5 FU缓释剂第1天的浸出液对人胰腺癌细胞株PC 3的抑制率达6 0. 2 7% ,第3天为3 4. 2 5% ,以后稳定在2 5. 0 0%左右。5 FU缓释剂瘤内注射治疗组裸鼠移植瘤生长速?

化疗及导向化疗治疗大肠癌与癌细胞凋亡研究

目的研究化疗及导向化疗对大肠癌组织及细胞的影响．方法大肠腺癌患者３７例，根据是否注射化疗药物分成Ａ，Ｂ两组．Ａ组１７例，术前不注射任何化疗药物或化疗药物的偶联物，将其作为对照组；Ｂ组２０例，其中７例手术前３ｄｉｖ丝裂霉素Ｃ（ＭＭＣ８ｍｇ），１３例手术前ｉｖ导向化疗药物抗ＣＥＡ单克隆抗体－ＭＭＣ交联物（含ＭＭＣ８ｍｇ，抗ＣＥＡ单克隆抗体１０ｍｇ）；其后对所有３７例患者行手术治疗并切取标本进行病理组织学观察，ＡｇＮＯＲ颗粒检测及肿瘤浸润淋巴细胞分类及计数．结果Ａ组病理退变程度１７例（１０００％）均为１级；Ｂ组病理退变程度１级４例（２００％），２级１２例（６００％），３级４例（２００％），两组之间差异有高度显著性（Ｐ＝０００００００３７６）．Ａ组癌细胞ＡｇＮＯＲ颗粒／核均数为４１１颗；Ｂ组癌细胞ＡｇＮＯＲ颗粒／核均数为２５８颗，其差异有高度显著性（Ｐ＜００１，ｔ＝５１４３）．肿瘤浸润淋巴细胞以Ｔ细胞ＣＤ＋３为主，同组中Ｔ淋巴细胞ＣＤ＋３（ａＰ＜００１，ｔ＝２０２２９）与Ｂ淋巴细胞ＣＤ＋２０之间的差异有高度显著性（ｂＰ＜００１，ｔ＝１９７６１）；Ａ组Ｔ淋巴细胞ＣＤ＋３为６０９１％，Ｂ组Ｔ?

细胞因子诱导的杀伤细胞降低乳腺癌耐药细胞系对阿霉素的耐受效应

目的 :探讨细胞因子诱导的杀伤细胞 (cytokine inducedkiller,CIK)与化疗药物对多药耐药 (multidrugre sistance,MDR)肿瘤细胞产生协同杀伤效应的机制。方法 :用细胞培养法加细胞因子在体外诱导并扩增CIK细胞 ,应用MTT法测定CIK细胞、阿霉素及二者联合对靶细胞的杀伤活性 ,用流式细胞术检测靶细胞上P糖蛋白 (P gp)的表达和细胞内药物积累 ,用细胞免疫组化方法检测P gp在MDR靶细胞MCF7adr内的分布。结果 :CIK细胞可使MCF7adr细胞P gp的表达活性下降 82 .5 % ,耐药靶细胞内阿霉素积累增加 3.2倍 ,使联合杀伤率比单纯加阿霉素(同等剂量 )组增加 7.8倍 ,比单纯加CIK细胞 (相同效靶比 )组增加 1.3倍。结论 :CIK细胞可明显抑制MCF7adr肿瘤细胞系的P gp表达 ,可协同阿霉素提高对MCF7adr肿瘤细胞系的杀伤活性。

肠胃清对人结直肠癌耐长春新碱细胞株HCT8／V的逆转作用

目的:探讨中药复方肠胃清对人结直肠癌耐长春新碱(vincristine,VCR)细胞株HCT8/V多药耐药的逆转作用。方法:采用细胞增殖抑制实验MTT法观察肠胃清药物血清对HCT8/V细胞株多药耐药性的逆转作用,采用高效液相色谱方法检测肠胃清药物血清作用后的耐药细胞内VCR药物浓度的改变。结果:本实验所采用的人结直肠癌耐VCR细胞株HCT8/V的耐VCR倍数为20.24倍,且对5-氟尿嘧啶、顺铂、羟基喜树碱和丝裂霉素这4种结构和作用机制各异的化疗药物有交叉耐药现象,具有多药耐药性,肠胃清能够逆转HCT8/V的耐药现象。高效液相色谱法结果显示,肠胃清药物血清处理HCT8/V48h后,细胞内的VCR浓度明显增高,且呈剂量依赖性。结论:肠胃清药物血清可提高HCT8/V细胞内化疗药物的浓度,有效逆转耐药细胞的多药耐药现象。

贝伐单抗联合化疗治疗晚期结直肠癌的临床观察

背景与目的:贝伐单抗(bevacizumab,BV)是抗血管内皮生长因子(vascular endothelialgrowth factor,VEGF)的人源化单抗,与化疗药物联用于治疗复发转移性结直肠癌,可明显改善疗效和生存期,不良反应可耐受。本文旨在观察贝伐单抗联合化疗治疗晚期结直肠癌的疗效及不良反应。方法:46例经病理组织学证实的晚期结直肠癌患者均接受贝伐单抗联合化疗治疗,贝伐单抗的剂量采用美国国立综合癌症网络(NCCN)推荐的5 mg/kg,每2周重复,或7.5 mg/kg,每3周重复。每2～3个疗程后评价疗效,同时观察并记录不良反应。结果:所有患者中,PR9例,SD30例,PD7例。客观有效率为20%,疾病控制率为85%。中位无进展生存期6.0个月,中位总生存期9.3个月。一线、二线、三线及以上应用贝伐单抗客观有效率分别为35%、6%和17%,疾病控制率分别为82%、88%和83%,中位无进展生存期分别为8.6、5.2和6.3个月,中位总生存期分别为12.2、8.3及8.1个月。不良反应为高血压3例,尿隐血阳性2例,蛋白尿1例,阴道出血1例,痰中带血1例,鼻衄1例,均为1～2级,对症治疗后均缓解:7例患者出现3～4级骨髓抑制,未出现粒缺性发热;2例患者因出现持续性2～4级肝功能受损未继续抗癌治疗。结论:贝伐单抗联合化疗治疗晚期结直肠癌的临床获益较肯定,且不良反应轻,患者耐受性较好,但需监测肝功能及血常规变化。