Tumor infiltrating lymphocytes(TILs) were isolated by enzymatic digestion and discontinuous gradient centrifugation from 8 human advanced tumors (4 stomach carcinoma, 2 liver cancer, 1 non-small-cell lung carcinoma and 1 colon cancer). These cells were cultured in complete RPMI 1640 medium supplemented with l000U/ml of rhIL2 for 4-6 weeks, till the cell number reach over l09/total, reinfused to the same patients i.v. meanwhile, the patients received 105U of rhIL2 i.m for 5 days. One week before and one month after TIL infusion periphery blood from the patients was collected and the mononuclear cells were isolated. Cytotoxicity against a panel of tumor cell targets by MTT colorometric assay and lymphocyte phenotype by two-color flow cytometry were mornitored. The results showed that there was significant increase in the killing ability to the tested tumor targets to different extent, especially the killing to the target cells which shared the same histological type with the patients tumor. (43 against 1249 lytic units p

Objective:To study the inhibitory effect of E1A gene on the growth of tumors in nude mice implanted with nasopharygeal carcinoma CNE2 cells and its promotion effect on the radiosensitivity of CNE2-implanted tumors,and to investigate the related mechanism.Methods: E1A gene was transfected into CNE2 cells using adenovirus system,and stable E1A positive clones were established.The inhibitory effect of E1A on tumor formation-ability of CNE2 cells was observed in nude mice.The efficacy of E1A gene therapy with or without radiotherapy against CNE2 cell-implanted tumors was evaluated.The effect of E1A gene therapy on the expression of P53 was detected by RT-PCR.Results: CNE2 cells stably transfected with E1A gene(CNE2-Ad-E1A)were successfully established.The tumor formation time was later and tumor size was smaller in CNE2-Ad-E1A cell-implanted mice compared with those in CNE2 cell-and CNE2-Ad-?-gal cell-implanted mice(CNE2 cells stably transfected with Ad-?-gal).Radiotherapy,E1A gene therapy and E1A gene+radiotherapy all suppressed the growth of implanted tumors,with the tumor suppression rates being(60.32?5.34)%,(70.53?6.12)%,and(97.15?4.87)%,respectively.E1A gene therapy significantly increased the expression of P53 gene in tumor tissues.Conclusion: E1A can inhibit the growth of tumors in mice implanted with nasopharygeal carcinoma cells,and enhance its sensitivity to radiotherapy,which may be related to the increased expression of P53 gene in tumor tissues.

Objective:To explore the inhibitory effects of gefitinib(epidermal growth factor receptor inhibitor) combined with celecoxib(cyclooxygenase-2 inhibitor) against human lung cancer A549 cells and the possible mechanism.Methods: A549 cells were cultured in RPMI 1640 medium and were divided into 4 groups: normal control group,5 ?mol/L gefitinib group,25 ?mol/L celecoxib group,and 5 ?mol/L gefitinib+25 ?mol/L celecoxib group.The morphological changes of A549 cells were observed under inverted microscope 48 h after treatment;the effects of drugs on growth of A549 Cells were detected by MTT assay;the apoptosis and cell cycles of A549 cells were measured by Annexin V/PI and Hoechst 33258 staining,respectively;and the expression of EGFR protein,COX-2 protein,and EGFR mRNA were determined by immunofluorescence and real-time PCR.Results: Compared with gefitinib and celecoxib groups,many granules and vacuoles were observed in the gefitinib and celecoxib combination group,and cells became round and there was defluxion.Both gefitinib and celecoxib inhibited the growth of A549 cells in a time-and dose-dependent manner.After treatment for 48 h,the inhibitory rate was(58.2?4.6)% in the combination group,which was significantly higher than those of the other two groups.Apoptosis rate in the combination group was also significantly higher than those in the other two groups(33.9% vs 6.0%,8.8%),and the cell proportion in S phase significantly decreased and in G0/G1 phases significantly increased(P

Objective:To investigate the inhibitory effect of Trichinella spiralis polypide protein on H7402 cell line in vitro and to assess its anti-tumor effect against hepatocellular carcinoma(HCC).Methods:The Trichinella spiralis polyp- ide protein was isolated from adult worms and newborn mixture.Trichinella spiralis polypide protein was prepared through homogenization and centrifugation,and its concentrations were determined by UV-visable spectrometers.H7402 cells trea- ted with Trichinella spiralis polypide protein(at 0.035,0.070,0.140 mg/ml)served as treatment group,and normal he- patic cell line HL-7702 treated with Trichinella spiralis polypide protein served as control group.Cell proliferation was de- tected by MTT;the effect of Trichinella spiralis polypide protein on migration and invasion ability of H7402 cell line was assessed by scarification test and invasion test.Cell apoptosis was observed by TUNEL.Apoptosis and cell cycle were ana- lyzed by FCM.Results:Trichinella spiralis polypide protein was successfully prepared.The protein at 0.035,0.070,0. 140 mg/ml inhibited H7402 cells proliferation by(22.40?13.80)%,(29.45?16.80)%,and(39.38?17.80)%, respectively.Apoptosis was observed by TUNEL and FCM,with an apoptosis rate of(39.07?0.90)%;the cell cycle was blocked at S phase.Scarification test and invasion test suggested that Trichinella spiralis polypide protein inhibited the migration and invasion of H7402 cell line,with the inhibition rate being(63.79?13.71)%.Conclusion:Trichinella spiralis polypide protein can inhibit the proliferation,migration,invision of H7402 cell line,and it might be a potential anti-cancer agent.

Objective:To investigate the enhancing effect of low dose adenovirus on rAAV2 expression in drug-sensi- tive and drug-resistant tumor cells and to explore the related mechanism.Methods:Human small cell lung cancer NCI- H446 cells,human lung adenocarcinoma A549 cells,human gastric cancer SGC7901 cells,human oral epithelial cancer KB cells,and VCR-resistant KB cells(KB/VCR)were transfected with rAAV2-GFP alone or combined with AdS-RFP or with AdS-TERT-RFP.GFP expression in the infected tumor cells was observed and analyzed by fluorescence microscope and fluorescence activated cell sorting(FACS).GFP protein and phosphorylation level of ERK and AKT in the infected tumor cells were detected by Western blotting.Quantitative analysis of DNA copies of GFP and mRNA expression of GFP, HSPG,?_v integrin,FGFR-1 in the infected tumor cells were performed by Real-time PCR.Results:The results of FACS demonstrated that the mean intensity of GFP fluorescence and the GFP positive rate after infection with rAAV2-GFP com- bined with Ad5-RFP or Ad5-TERT-RFP increased by 0.3-3 and 4-8 folds,respectively.GFP expression in the tumor cells with combined infection showed 4-6 folds increase and the phosphorylation level of ERK and AKT also increased.GFP mRNA expression in the combined infection tumor cells increased by 3.83-7.33 folds;there was no differences in the GFP DNA copies between rAAV2-GFP group and rAAV2-GFP plus Ad5-RFP group.Cellular receptors HSPG,?_v integrin and FGFR-1 mRNA expression was slightly increased in rAAV2-GFP plus Ad5-RFP.Conclusion:Lower dose of recombinant adenovirus can obviously enhance the expression of rAAV2 in drug-sensitive and drug-resistant tumor cells,which might be related to the activation of signal transduction pathway and increase of intracellular transcription.

Objective: To determine the inhibitory effects of anti-bone sialoprotein(BSP) antibody on human breast cancer cells adhering to bone matrix in vitro.Methods: Expression of BSP in 3 different cancer cell lines(bone seeking breast cancer cell MDA-MB-231-BO,lung adenocarcinoma cell SPCA-1 and colon carcinoma cell LOVO)was detected immunohistochemically.Cells adhering test was carried out to investigate the adhering of the 3 different cancer cell lines to mouse bone matrix in vitro and the inhibitory effect of anti-BSP antibody on MDA-MB-231-BO cells adhering to mouse bone matrix;Enzyme-linked immunosorbent assay was carried out for quantitative detection of TGF-?.Results: BSP immunostaining was positive in MDA-MB-231-BO cells and negative in SPCA-1 and Lovo cells.The number of MDA-MB-231-F10 cells adhering to mouse bone matrix was significantly more than SPCA-1 or Lovo cells(P

Objective: To investigate the role of Bcl-XL siRNA in sensitizing ovary cancer cells for TRAIL-induced apoptosis and the underlying mechanisms.Methods: Western blotting analysis was performed to confirm whether Bcl-XL siRNA could effectively down-regulate Bcl-XL protein after SKOV3 cells were transfected with Bcl-XL siRNA and then the cells were treated with TRAIL.Flow cytometry analysis and cell counting were used for assessment of apoptotic rate and survival rate,respectively.Western blotting analysis was performed to determine the changes of apoptosis-related protein in SKOV3 cells.Results: Compared with control group,Bcl-XL siRNA transfection effectively down-regulated the expression of Bcl-XL protein and suppressed the growth of SKOV3 cells;the suppression peaked at 96 h after transfection,being 43.9% that of the control group(P

Objective: To investigate the relationship between the expression of Raf-1 and tumor angiogenesis,and to discuss its clinical significance.Methods: Tissue microarray technique was used to detect the expression of Raf-1 in 87 specimens of human colon carcinoma,their corresponding adjacent tissues,and incision margins.The patients were from the Department of Pathology of Xijing Hospital between 2005 and 2006.Microvessel density(MVD) was detected using immunohistochemistry with CD34 labeling.The correlation between Raf-1 expression and MVD with the tumor size,metastasis,and differentiation was analyzed.Results: The positive rates of Raf-1 in colon carcinoma tissues,adjacent tissues and incision margins were 86.47%,37.34% and 11.03%,respectively;there were significant difference among the 3 values(P

Objective: To observe the safety and clinical efficacy of tumor antigen-pulsed dendritic cell(DC) vaccine in treatment of advanced malignant tumor.Methods: Ninety-one patients with non-small cell lung cancer,colon and rectal cancer,melanoma,renal carcinoma,breast cancer and other malignant tumors were enrolled in this study.All patients met the selecting standard and signed informed consent.Human dendritic cells were obtained from peripheral blood monocytes by culturing them with granulocyte macrophage-colony stimulating factor and interleukin-4.DC vaccine was prepared from tumor antigen pulsed immature dendritic cells in vitro.Patients received the vaccine therapy once every week and one cycle was defined as once every week for 3 weeks.Results: All the patients received 96 cycles of DC vaccine treatment.Symptoms of toxicity included fever,shivering,aching pain of muscle,asthenia,itching,stifle and transient fatigue;most of the symptoms automatically recovered.Clinical efficacy of the treatment was evaluated in 76 patients.Thirty-one of the 76 patients were stable after treatment and 45 were in progressive situation,with the clinical benefiting rate being 40.8%.Eighty-five patients were followed up.The median time for progression was 2.6 months;the overall survival time was 0.9-30.6 months;and the median survival period was 4.5 months,with the one year survival rate being 9.2%.Conclusion: The results suggest that the DC vaccine therapy is well tolerated in treating patients with advanced malignant tumors and has satisfactory clinical benefit;the clinical value of DC vaccine therapy needs to be further observed.

DC group.IL-12 secretion in IL-18/fusion group was higher than that in the fusion group,and IL-12 in the pulsed DC group was higher than that in the DC group.The in vitro killing rates of the 4 groups were 79.73%,50.68%,35.81% and 4.05%,respectively.Tumor forming time in IL-18/fusion group([12.82?2.85]d) was longer than those in the pulsed DC group([8.52?1.97]d,P