1.Serial Analysis of Gene Expression in Immortalized BEP2D Cells and Malignant Transformed BEP2D Cells Induced by α-particle
Shi-Li GE ; Gang LI ; Wei CHEN ; Tie-Zhu LOU ; De-Chang WU
Chinese Journal of Cancer 2001;20(1):12-17
Objective: The authors used the serial analysis of gene expression (SAGE) method to analyze transcripts present in the immortalized BEP2D cells and the malignant transformed BEP2D cells. Methods: As a step toward understanding the complex gene expression differences between the immortalized BEP2D cells and the malignant transformed BEP2D cells induced by α particle,SAGE method was introduced into this experiment. Technological methods included total RNA extraction, mRNA isolation, full length of dscDNA synthesis, PCR, transformation and sequencing. SAGE SoftWare was used to analyze tag sequences and compare the abundance of tags between two SAGE libraries. Results: Two independent SAGE libraries were constructed from the immortalized BEP2D cells and the malignant transformed BEP2D cells induced by 1.5 Gy α particles. A total of 2 331 SAGE tags were identified for the sequences, representing 252 unique transcripts. Though up to now the obtained information is limited, comparison of the two SAGE libraries indicated a remarkable similarity in the expression profiles. Of the 252 transcripts detected, 12 transcripts (4.8% ) matched no reliable known genes in UniGene library. Combination of Northern blot hybridization, the expression level of TGF β induced Smad7 gene in the malignant transformed cells was higher than that in the immortalized cells. Conversely, Chemokine receptor CCR11 gene was expressed at lower levels in the malignant transformation cells. Conclusion: (1) Out of results given by SAGE,the tendency of the abundance of gene expression and the gene expression differentiation between two cell lines were described. (2) The expression level of TGF β induced Smad7 gene in the malignant transformed BEP2D cells was higher than that in the immortalized cells and chemokine receptor CCR11 gene was expressed at lower levels in the malignant transformed BEP2D cells. (3)SAGE was a powerful method in a quantitative and simultaneous analysis of a large number of transcripts in any particular cell system, especially in defining functions of known genes.
2.Tumor antigen-pulsed dendritic cell vaccine for treatment of patients with advanced malignant tumor:a clinical observation
Liming ZHU ; Yiping ZHANG ; Xiaojia WANG ; Haijun ZHONG ; Yun FAN ; Xinmin YU ; Qinghua DENG ; Xiangming KONG ; Jieer YING ; Tie LI ; Guangyuan LOU ; Zhibing WU ; Zhongzhu TANG ; Shengling MA
Chinese Journal of Cancer Biotherapy 2006;0(06):-
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.