Bevacizumab plus infusional 5-fluorouracil,leucovorin and irinotecan for advanced colorectal cancer that progressed after oxaliplatin and irinotecan chemotherapy:A pilot study

AIM： To evaluate the combination of bevacizumab with infusional 5-fluorouracil （5-FU）, leucovorin （LV） and irinotecan （FOLFIRI） in patients with advanced colorectal cancer （CRC） pretreated with combination regimens including irinotecan and oxaliplatin. METHODS： Fourteen patients （median age 56 years） with advanced CRC, all having progressed after oxaliplatin- and irinotecan-based combination chemotherapy, were enrolled in this study. Patients were treated with 2 h infusion of irinotecan 150 mg/m2 on d 1, plus bevacizumab 5 mg/kg iv infusion for 90 min on d 2, and iv injection of LV 20 mg/m2 followed by a bolus of 5-FU 400 mg/m2 and then 22 h continuous infusion of 600 mg/m2 given on two consecutive days every 14 d. RESULTS： The median number of cycles of chemotherapy was six （range 3-12）. The response rate was 28.5%, one patient had a complete response, and three patients had a partial response. Eight patients had stable disease. The median time to progression was 3.9 mo （95% CI 2.0-8.7）, and the median overall survival was 10.9 mo （95% CI 9.6-12.1）. Grade 3/4 neutropenia occurred in five patients, and two of these developed neutropenic fever. Grade 3 hematuria and hematochezia occurred in one. Grade 2 proteinuria occurred in two patients. However, hypertension, bowel perforation or thromboembolic events did not occur in a total of 90 cycles. CONCLUSION： Bevacizumab with FOLFIRI is well tolerated and a feasible treatment in patients with heavily treated advanced CRC.

Oxaliplatin,Fluorouracil and Leucovorin (FOLFOX) as First-line Chemotherapy for Metastatic or Recurrent Colorectal Cancer Patients

OBJECTIVE To investigate the efficiency and safety of the oxaliplatin, fluorouracil （5-FU） and leucovorin regimen （FOLFOX）in previously untreated patients with metastatic or recurrent colorectal cancer. METHODS Previously untreated patients with metastatic or recurrent colorectal cancer received 100 mg/m^2 of oxaliplatin intravenously （Ⅳ） over 2 h on day 1, and Ⅳ 400 mg/m^2 of leucovorin over 2 h followed by a bolus of 400 mg/m^2 of 5-FU. Then 2,600-3,000 mg/m^2 of 5-FU was administered by continuous infusion over 46 h. RESULTS An evaluated response rate was determined for 97 of 105 treated patients. The overall response rate was 35.1%, 9 patients （9.3%） had a complete response and 25 patients （25.8%） a partial response. Thirtytwo patients （33.0%） developed stable disease and 32.0% of the patients progressed. The median time to progression （TTP） was 7.7 months and the median overall survival 20.5 months. One and 2-year survival rates were 68% and 32%. Toxic effects based on the National Cancer Institute-Common Toxicity Criteria （NCI-CTC）, reaching grade 3/4 were： neutropenia 12.3%, anemia 11.3%, vomiting 4.1% and diarrhea 7.2%. Grade 3 neuropathy was 5.1%. The overall survival rate of patients who had received a radical resection was superior to the patients who had not received a operation, or had received a palliative resection （P=0.0658）. The serum levels of CEA, ALP and LDH had no relationship with survival （P〉0.05）. CONCLUSION The FOLFOX regimen containing oxaliplatin, 5-FU plus leucovorin was an efficacious regimen with good tolerability in previously untreated metastatic or recurrent colorectal cancer patients.

Oxaliplatin combined with 5-fluorouracil, leucovorin regimen for patients with advanced colorectal cancer

Objective： To observe the efficacy and tolerability of continuously infusing 5-fluorouracil （5-FU） / folic acid combined with oxaliplatin （L-OHP/5-FU/LV regimen） as first line treatment in advanced colorectal cancer. Methods： 23 patients of advanced colorectal cancer were treated with 5-FU 500 mg/d, civ, d 1-d5, d8-d12, leucovorin 100 mg/d, iv gtt, d1, d8, folic acid tablet 60 mg/d, po, d2-d5, d9-d12, and oxaliplatin 65 mg/（m^2·d）, iv gtt, dl, d8, repeated every 21 days （one cycle）. The effect was evaluated after two cycles. Results： Complete response in 2 cases and partial response in 10 cases were observed with an overall response rate of 47.18%. Adverse effects were mainly grade 1-2, including nausea, vomiting, diarrhea, dental ulcer, peripheral neuritis and myelosuppression. Conclusion： L-OHP/5-FU/LV regimen is an effective and better tolerated alternative treatment in advanced colorectal cancer and yields promising clinical application.

Effects of oxaliplatin, leucovorin and fluorouracil on serum tumor markers, VEGF, CRP and matrix metalloproteinases in patients with advanced esophageal cancer

Objective: To investigate the effects of oxaliplatin, leucovorin and fluorouracil on serum tumor markers, VEGF, CRP and matrix metalloproteinases in patients with advanced esophageal cancer. Methods: From March 2012 to March 2017 a total of 248 patients with advanced esophageal cancer were selected as the study subjects. According to random data table, they were divided into control group (n=123) and observation group (n=125) according to random data table. The control group was treated with cisplatin combined with fluorouracil, leucovorin chemotherapy, and patients in the observation group received oxaliplatin, leucovorin and fluorouracil chemotherapy, all patients were treated for 2 cycles. The changes of serum tumor markers, VEGF, CRP and matrix metalloproteinase levels in the two groups before and after treatment was compared. Results: Before treatment, there was no significant difference of the levels of serum CA125, CA19-9, CEA, VEGF, CRP, MMP-2 and MMP-9 between the control group and the observation group. Compared with the group before treatment, the levels of CA125, CA19-9, CEA, VEGF, CRP, MMP-2 and MMP-9 in the two groups were significantly lower. After treatment, the level of CA125, CA19-9, CEA, VEGF, CRP, MMP-2 and MMP-9 in the observation group was significantly lower than those of the control group. Conclusion:Oxaliplatin, leucovorin and fluorouracil chemotherapy can effectively reduce the levels of serum tumor markers, VEGF, CRP and matrix metalloproteinase in patients with advanced esophageal cancer, it has important clinical value.

Value of Myocardial Strain in Monitoring Fluorouracil-Based Chemotherapy-Related Cardiac Dysfunction in Gastrointestinal Cancer Patients

Objective To investigate the predictive value of myocardial strain for cardiotoxicity associated with fluorouracil-based chemotherapies in gastrointestinal cancer patients.Methods Patients with diagnosis of gastrointestinal cancers,who were hospitalized for chemotherapy involving antimetabolic drugs,were eligible in this prospective study.Echocardiography was performed before and after each chemotherapy cycle during hospitalization until the completion of chemotherapy.Cancer therapy-related cardiac dysfunction(CTRCD)was identified if there was a decrease in left ventricular ejection fraction(LVEF)by at least 5%to an absolute value of<53%from the baseline,accompanied by symptoms or signs of heart failure;or a decrease in LVEF of at least 10%to an absolute value of<53%from the baseline,without symptoms or signs of heart failure.Subclinical cardiac impairment is defined as a decrease in the left ventricular global longitudinal strain(GLS)of at least 15%from baseline.Clinical data and myocardial strain variables were collected.Changes of echocardiographic indexes after chemotherapy at each cycle were observed and compared to those of pre-chemotherapy.Cox regression analysis was used to determine the associated indexes to CTRCD,and receiver operating characteristic(ROC)curves were plotted for evaluation of their predicting efficacy.Results Fifty-one patients completed 4 cycles of chemotherapy and were enrolled in the study analysis.LVEF,GLS,GLS epicardium(GLS-epi),and GLS endocardium(GLS-endo)were decreased after the 4 cycles of chemotherapy.Throughout the chemotherapy period,6 patients(11.8%)progressed to CTRCD.The Cox regression analysis revealed that the change in left atrial ejection fraction(LAEF)and LAS during the reservoir(LASr)phase after the first cycle of chemotherapy(C1v-LAEF and C1v-LASr,respectively)were significantly associated with the development of CTRCD[C1v-LAEF(HR=1.040;95%CI:1.000-1.082;P=0.047);C1v-LASr(HR=1.024;95%CI:1.000-1.048;P=0.048)].The sensitivity and specificity were 50.0%and 93.3%,respectively,for C1v-LAEF predicting CTRCD when C1v-LAEF>19.68%was used as the cut-off value,and were 66.7%and 75.6%,respectively,for C1v-LASr predicting CTRCD when C1v-LASr>14.73%was used as the cut-off value.The areas under the ROC curve(AUC)for C1v-LAEF and C1v-LASr predicting CTRCD were 0.694 and 0.707,respectively.Conclusion GLS changes among patients with subclinical impairment of cardiac function who were treated with fluorouracil-based chemotherapies,and C1v-LAEF and C1v-LASr of the left atrium are early predictors of cardiac function deterioration.

肿瘤阳性显像剂5-^(18)F-fluorouracil的标记与动物实验

目的 探讨18F fluorouracil(FU)作为肿瘤阳性显像剂的可能性。方法 标记与合成了18F FU ,研究其在正常与荷瘤裸鼠中的生物分布 ,进行正常与荷瘤兔的PET显像。结果 HPLC及其他质控分析结果均证实18F FU动物实验与临床应用的可行性 ,生物分布及显像研究表明它在肿瘤组织中有较多的摄取。结论 18F FU是一种有潜力的肿瘤阳性显像剂。

亲肿瘤显像剂5-^(18)F-Fluorouracil(^(18)F-FU)的标记合成研究

用18 F F2 与Uracil的直接标记法合成了 5- 18F -Fluorouracil,探索18F -FU -PET显像对结肠直肠癌肝转移病人化疗前后疗效评价的意义。产品的放射化学纯 >95% ,放射性得率为 60 % ,pH值为 7,无菌性试验和热源试验均为阴性 ,这一结果为临床应用提供了保证。

PhaseⅡstudy of oxaliplatin combined with S-1 and leucovorin(SOL)for Chinese patients with metastatic colorectal cancer

Background:Fluoropyrimidine and oxaliplatin are widely used for patients with colorectal cancer.This phase II study was conducted to evaluate the efficacy and safety of the combination of S-1,oxaliplatin,and leucovorin(SOL) in the treatment of Chinese patients with metastatic colorectal cancer(mCRC).Methods:Eligible patients with untreated mCRC from four hospitals in China received intravenous oxaliplatin(85 mg/m2) on day 1,oral S-1 twice daily(80-120 mg per day) on day 1-7,and leucovorin twice daily(50 mg per day)simultaneously with S-1,every 2 weeks.Results and discussion:Forty patients were enrolled in our study.In total,296 cycles of SOL were administered.The overall response rate was 50.0%.At a median follow-up of 27 months,progression-free survival and overall surviva were 7.0 months(95%confidence interval[CI]6.0-10.6 months) and 22.2 months(95%CI 15.1-29.3 months),respectively.The most common grade 3/4 non-hematological adverse events were diarrhea(n = 8,20.0%),nausea(n = 3,7.5%),and vomiting(n = 3,7.5%).The most common grade 3/4 hematological toxicities were thrombocytopenia(n = 3,7.5%),neutropenia(n = 1,2.5%),and abnormal alanine transaminase/aspartate transaminase levels(n = 1,2.5%).There was one treatment-related death.Conclusions:The data indicate that the SOL regimen is effective and moderately tolerated in Chinese patients with mCRC.Trial registration:Clinical trial

Interaction between cisplatin,5-fluorouracil and vincristine on human hepatoma cell line (7721)

AIM To evaluate the killing effects of CDDP, 5-Fu and VCR on human hepaoma cell line (7721).METHODS The median-effect principle was used.RESULTS Killing effects of the individual drug were enhanced as the median concentration increased. Antagonism was produced when two drugs were used at a higher concentration (CI＞1), and synergism was achiened when CI＜1. Finally, the effect was influenced by both the ratios of drug concentration and the sequence of administration.CONCLUSION The drug administration order and drug concentrations are significant factors that need to be considered in clinical practice.INTRODUCTIONThe combined chemotherapy for malignant carcinoma is desired to produce efficacious synergism between each drug, alleviate side effects of drugs and delay drug resistance. Clinically, the interaction (namely synergism, summation and antagonism) of different anticancer drugs in combination is usually evaluated by Chou-Talalay′s combination index (i.e., median-effect principle)[1-9]. In this paper the combination effect between Cisplatin (Cis), 5-Fluorouracil (5-Flu) and Vincristine (VCR) on human hepatoma cell line 7721, was analyzed in vitro.

Preparation and distribution of 5-fluorouracil ^(125)I sodium alginate-bovine serum albumin nanoparticles

AIM To prepare 5 FU sodium alginate 125 I bovine serum albumin nanoparticles (BSA NP), to determine the radioactive count in different organs of rats at different time points after oral administration of 5 FU 125 I sodium alginate BSA NP and to calculate the kinetic parameters of its metabolism. METHODS Emulsion solidification method was used to prepare 5 FU 125 I sodium alginate BSA NP, and to determine its diameter under transmission electronic microscope (TEM). Then the rate of NP and external drug releasing velocity were measured. Radioactive counting in different organs of rats was made after oral administration of the NP by GAMA Counter, and the kinetic parameters of drug metabolism were calculated by handling the data with the two department model. RESULTS The average arithmatic diameter of the NP was 166nm ± 34nm , the rate of 5 FU was 32 8% and the cumulative external releasing ratio amounted to 84 0% within 72 hours. The NP was mainly distributed in the liver, spleen, lungs and kidneys after NP oral administration to rats. The micro radioautographic experiment showed that NP was distributed in the Kupffers cells of liver, liver parenchymal cells and the phagocytes of spleen and lungs. The kinetic parameters of matabolism were: T 1/2 =9 42h, C max =2 45×10 7Bq, T max =2 18h, AUC=148×10 9Bq. CONCLUSION NP is difficult to pass through the blood-cerebral barrier,and 125 I sodium alginate-BSA NP enters the body circulation by gastroin testinal passage.

Rosiglitazone enhances fluorouracil-induced apoptosis of HT-29 cells by activating peroxisome proliferator-activated receptor γ

AIM： To examine whether and how rosiglitazone enhances apoptosis induced by fluorouracil in human colon cancer （HT-29） cells. METHODS： Human colon cancer HT-29 cells were cultured in vitro and treated with fluorouracil and/or rosiglitazone. Proliferation and growth of HT-29 cells were evaluated by MTF assay and trypan blue exclusion methods, respectively. The apoptosis of HT-29 cells was determined by acridine orange/ethidium bromide staining and flow cytometry using PI fluorescence staining. The expressions of peroxisome proliferator-activated receptor γ （PPARγ）, Bcl-2 and Bax in HT-29 cells were analyzed by Western blot. RESULTS： Although rosiglitazone at the concentration below 30 μmol/L for 72 h exerted almost no inhibitory effect on proliferation and growth of HT-29 cells, it could significantly enhance fluorouracil-induced HT-29 cell proliferation and growth inhibition. Furthermore, 10 μmol/L rosilitazone did not induce apoptosis of HT-29 cells but dramatically enhanced fluorouracil-induced apoptosis of HT-29 cells. However, rosiglitazone did not improve apoptosis induced by fluorouracil in HT-29 cells pretreated with GW9662, a PPARγ antagonist. Meanwhile, the expression of Bax and PPAR7 was upregulated, while the expression of Bcl-2 was down regulated in HT-29 cells treated with rosiglitazone in a time-dependent manner. However, the effect of rosiglitazone on Bcl-2 and Bax was blocked or diminished in the presence of GW9662. CONCLUSION： Rosiglitazone enhances fluorouracilinduced apoptosis of HT-29 cells by activating PPARγ.

UGT1A1 predicts outcome in colorectal cancer treated with irinotecan and fluorouracil

AIM:To evaluate effects of UDP-glucuronosyltransferase1A1(UGT1A1) and thymidylate synthetase(TS) gene polymorphisms on irinotecan in metastatic colorectal cancer(mCRC).METHODS:Two irinotecan-and fluorouracil-based regimens,FOLFIRI and IFL,were selected as second-line therapy for 138 Chinese mCRC patients.Genomic DNA was extracted from peripheral blood samples before treatment.UGT1A1 and TS gene polymorphisms were determined by direct sequencing and restriction fragment length polymorphism,respectively.Gene polymorphisms of UGT1A1*28,UGT1A1*6 and promoter enhancer region of TS were analyzed.The relationship between genetic polymorphisms and clinical outcome,that is,response,toxicity and survival were assessed.Pharmacokinetic analyses were performed in a subgroup patients based on different UGT1A1 genotypes.Plasma concentration of irinotecan and its active metabolite SN-38 and inactive metabolite SN-38G were determined by high performance liquid chromatography.Differences in irinotecan and its metabolites between UGT1A1 gene variants were compared.RESULTS:One hundred and eight patients received the FOLFIRI regimen,29 the IFL regimen,and one irinotecan monotherapy.One hundred and thirty patients were eligible for toxicity and 111 for efficacy evaluation.One hundred and thirty-six patients were tested for UGT1A1*28 and *6 genotypes and 125 for promoter enhancer region of TS.Patients showed a higher frequency of wild-type UGT1A1*28(TA6/6) compared with a Caucasian population(69.9% vs 45.2%).No significant difference was found between response rates and UGT1A1 genotype,although wild-type showed lower response rates compared with other variants(17.9% vs 24.2% for UGT1A1*28,15.7% vs 26.8% for UGT1A1*6).When TS was considered,the subgroup with homozygous UGT1A1*28(TA7/7) and non-3RG genotypes showed the highest response rate(33.3%),while wild-type UGT1A1*28(TA6/6) with non-3RG only had a 13.6% response rate,but no significant difference was found.Logistic regression showed treatment duration was closely linked to clinical response.In toxicity comparison,UGT1A1*28 TA6/6 was associated with lower incidence of grade 2-4 diarrhea(27.8% vs 100%),and significantly reduced the risk of grade 4 neutropenia compared with TA7/7(7.8% vs 37.5%).Wild-type UGT1A1*6(G/G) tended to have a lower incidence of grade 3/4 diarrhea vs homozygous mutant(A/A) genotype(13.0% vs 40.0%).Taking UGT1A1 and TS genotypes together,lower incidence of grade 2-4 diarrhea was found in patients with non-3RG TS genotypes,when TA6/6 was compared with TA7/7(35.3% vs 100.0%).No significant association with time to progression(TTP) and overall survival(OS) was observed with either UGT1A1 or TS gene polymorphisms,although slightly longer TTP and OS were found with UGT1A1*28(TA6/6).Irinotecan PK was investigated in 34 patients,which showed high area under concentration curve(AUC) of irinotecan and SN-38,but low AUC ratio(SN-38G /SN-38) in those patients with UGT1A1*28 TA7/7.CONCLUSION:A distinct distribution pattern of UGT1A1 genotypes in Chinese patients might contribute to relatively low toxicity associated with irinotecan and 5-fluorouracil in mCRC patients.

Effects of tacrolimus on proliferation, apoptosis, and fluorouracil sensitivity of liver cancer cell line of SMMC-7721

BACKGROUND: The effect of tacrolimus (FK506) and 5- fluorouracil (5-FU) on hepatocellular carcinoma remains elusive. The aim of this study was to assess the effect of ta- crolimus on the proliferation, and apoptosis in liver cancer cell line of SMMC-7721 and its sensitivity to fluorouracil (5- FU). METHODS: The liver cancer cell line of SMMC-7721 was cultured in vitro, and the MTT assay was used to examine the antiproliferative effect of FK506. Flow cytometry (FCM) was used to examine the effect of 5-FU alone or in combination with FK506 on the apoptosis and cell cycle of SMMC-7721 cells. RESULTS: FK506 produced concentration-dependent anti- proliferative effect on SMMC-7721 cells at all experimental concentrations(P <0.05), but no effect on induction of apo- ptosis. 5-FU induced apoptosis in a concentration-depen- dent manner, whereas the percentage of G0/G1-phase cells and proliferation index (PI) were increased with the in- creased concentration of 5-FU. Pretreatment with FK506 for 2 hours enhanced the effect of 5-FU on the induction of apoptosis. CONCLUSIONS: FK506 inhibits the growth of SMMC-7721 cells and enhances their sensitivity to 5-FU. This may be as- sociated with the synergic effect of FK506 and 5-FU in in- ducing apoptosis and G0/G1-phase stasis.

Caspase-8 in apoptosis of hepatoma cell induced by 5-fluorouracil

OBJECTIVE: To explore the relationship between the changes in the activity of caspase-8 and apoptosis of HepG2 cells induced by 5-fluorouracil (5-Fu). METHODS: Human hepatoma HepG2 cells were treated with 5-Fu at the concentrations of 1×10^(-1), 1×10^(-2), 1×10^(-3), 1×10^(-4), 1×10^(-5) mol/L and for 1, 2, 4, 8, 16, 24 hours, respectively. The caspase-8 activity was detected using caspase-8 fluorescent assay kit. The apoptotic rate of HepG2 cells induced by 5-Fu with or without the caspase-8 inhibitor IETD-FMK was measured by flow cytometry. RESULTS: Afer the HepG2 cells were trealed with 10^(-2) mol/L 5-Fu, the caspase-8 activity increased gradually and reached the peak level (313.9±6.9) at 16 hours, then fell down. Compared with the control group, the activity was still significantly higher (274.2±3.9 vs 68.3±3.6, P<0.01). With the increasing concentraion of 5-Fu, the caspase-8 activity was also increased; the activity in high concentration 5-Fu was significantly higher than that in low concentration 5-Fu (370.5±4.7 vs 313.7±6.9; 225.7±5.4 vs 183.3±4.8; 183.3±4.8 vs 124.0±6.2, P<0.01). The caspase-8 activity was the highest at 1×10^(-1) mol/L 5-Fu (370.5±4.7). The caspase-8 activity in low concentration 5-Fu was higher than in the blank control group and inhibitor group (124.0±6.2 vs 68.5±3.4; 124.0±6.2 vs 41.0±2.1, P<0.01). IETD-FMK could block the activation of caspase-8 and reduce the apoptosis of HepG2 cells induced by 5-Fu. The apoptotic rate of HepG2 cells in the 5-Fu group was significantly different from that in the inhibitor group (P<0.01). CONCLUSIONS: 5-Fu can induce apoptosis of HepG2 cells via caspase-8 signal transduction pathway, which can be blocked by IETD-FMK. 5-Fu promotes the increase of caspase-8 activity in a time- or concentration-dependent manner.

Pneumatosis cystoides intestinalis after fluorouracil chemotherapy for rectal cancer

Pneumatosis cystoides intestinalis (PCI) is a relatively rare condition characterized by intraluminal gas in the gastrointestinal tract. Several chemotherapeutic agents have been reported to be associated with PCI, although fluorouracil-related PCI is extremely rare. We report a case of a 76-year old man who received adjuvant chemotherapy for rectal cancer with fluorouracil (FU) and leucovorin (LV). After 1 cycle of the treatment, he presented with diarrhea and abdominal pain. Abdominal radiogram revealed the presence of free air under the diaphragm and intramural gas in the intestine. Laparotomy was performed, showing a suspected diagnosis of perforation in the gastrointestinal tract. Intraoperative findings revealed penumatosis of the intestine without evidence of perforation. He was treated supportively and his symptoms improved. In conclusion, we should consider the possibility of PCI occurring in patients with malignancies during chemotherapy treatment.

Pharmacological Study on Antitumor Activity of 5-Fluorouracil-1-Acetic Acid and Its Rare Earth Complexes

The antitumor activity of 5 fluorouracil 1 acetic acid(HFAA) and its lanthanide complexes(La(FAA) 3, Eu(FAA) 3) were studied. The results show that HFAA, La(FAA) 3 and Eu(FAA) 3 with the concentrations of 1 0×10 -5 ～1 0×10 -2 μg·ml -1 inhibit the colony formation of leukemia cells(L 1210 ) and the growth of transplanted tumor sarcoma 180(S 180 ), hepatic carcinoma(HEPA) and ehrlich ascites tumor(EC) as well. The maximum inhibitory rate of Eu(FAA) 3 for S 180 is 38 4%, that HFAA and La(FAA) 3 for EC are 22 4% and 43 4%, respectively. The life prolongation rate of Eu(FAA) 3 for HEPA bearing mice is as long as 284%.

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.

Rosiglitazone enhances 5-fluorouracil-induced cell growth inhibition in hepatocellular carcinoma cell line Hep3B

Background and Objective: Rosiglitazone is a peroxisome proliferators-activated receptor γ (PPARγ) ligand, which inhibits tumor growth by activating PPARγ signaling pathways. Fluorouracil (5-FU) is one of the commonly used chemotherapeutic drugs. However, patients develop drug resistance of 5-FU over time. The aim of this study was to investigate whether rosiglitazone can enhance 5-FU-induced cell growth inhibition and to explore its potential mechanisms.Methods: Cell viability was measured using MTT assay. Protein expression levels were detected by Western blot analysis. Small interference RNA was utilized to knockout PPARγ and PTEN in Hep3B cells. Results: After 48 h of treatment with 10, 20, and 40 μmol/L rosiglitazone, the viability of Hep3B cells was (78.0 ± 2.7)%, (37.3 ± 8.1)%, and (19.8 ± 2.2)%, respectively (compared with control group, P values were all < 0.001). After 48 h of treatment with 10 μmol/L 5-FU, the viability of Hep3B cells was about (82.6 ± 3.9)%. When cells were treated with 10 μmol/L 5-FU in combination with either 10, 20 or 40 μmol/L rosiglitazone, the cell viability was (51.6 ± 5.4)%, (14.8 ± 4.2)%, and (8.5 ± 0.9)%, with corresponding q value of 1.36, 1.23, and 1.19, respectively. These data suggested that the two drugs had synergic effect in inhibiting Hep3B cell growth, which was further confirmed in an in vivo mice model. Subsequent investigations showed that rosiglitazone activated PPARγ signaling pathways and increased the expression of PTEN. Conclusions: Rosiglitazone enhances 5-FU-induced cell growth inhibition of Hep3B cells.

Cooperative inhibitory effect of sinomenine combined with5-fluorouracil on esophageal carcinoma

AIM:To investigate the inhibitory effects of sinomenine(SIN)combined with 5-fluorouracil(5-FU)on esophageal carcinoma in vitro and in vivo.METHODS:Esophageal carcinoma(Eca-109)cells were cultured in DMEM.The single or combined growth inhibition effects of SIN and 5-FU on the Eca-109 cells were examined by measuring the absorbance of CCK-8dye in living cells.Hoechst 33258 staining and an Annexin V/PI apoptosis kit were used to detect the percentage of cells undergoing apoptosis.Western blotting was used to investigate the essential mechanism underlying SIN and 5-FU-induced apoptosis.SIN at 25mg/kg and 5-FU at 12 mg/kg every 3 d,either combined or alone,was injected into nude mice and tumor growth inhibition and side effects of the drug treatment were observed.RESULTS:SIN and 5-FU,both in combination and individually,significantly inhibited the proliferation of Eca-109 cells and induced obvious apoptosis.Furthermore,the combined effects were greater than those of the individual agents(P<0.05).Annexin V/PI staining and Hoechst 33258 staining both indicated that the percentage of apoptotic cells induced by SIN and 5-FU combined or alone were significantly different from the control(P<0.05).The up-regulation of Bax and downregulation of Bcl-2 showed that the essential mechanism of apoptosis induced by SIN and 5-FU occurs via the mitochondrial pathway.SIN and 5-FU alone significantly inhibited the growth of tumor xenografts in vivo,and the combined inhibition rate was even higher(P<0.05).During the course of chemotherapy,no obvious side effects were observed in the liver or kidneys.CONCLUSION:The combined effects of SIN and 5-FU on esophageal carcinoma were superior to those of the individual compounds,and the drug combination did not increase the side effects of chemotherapy.