Journal Detail Information

1

Cite

Share

Construction and expression of extracellular domain of human death receptor 5 and detection of its biological activity

Changgong ZHANG ; Yuguo SONG ; Wenzhu LI ; Li WANG ; Caixia CHEN ; Guohong ZHUANG

Chinese Journal of Cancer Biotherapy.2006;0(06):-.

Objective: To construct the expressing vector of the extracellular domain of death receptor 5 (DR5), express it E.coli, identify the purified DR5 protein, and study its biological activity. Methods: The extracellular domain of DR5(eDR5) was assembled by overlapping PCR. The expression vector pET-22b(+)/ DR5 was constructed and transformed into E.coli BL21(DE3). The expression of eDR5 protein was induced by IPTG and purified by Ni 2+ -affinity chromatographic column. The purity and specificities were detected by SDS-PAGE and ELISA, respectively. The blocking effects of purified eDR5 on FMU1.5-induced apoptosis of U343, U373 cells were observed. Results: The extracellular domain of DR5 was obtained by overlapping PCR. The eDR5 protein was expressed in both supernatants and inclusion bodies with a yield more than 30% of total bacterial proteins. The purity of eDR5 was more than 95% and the yield reached 9 mg/ml. The result of ELISA showed the purified protein was eDR5. Purified eDR5 partially blocked the apoptosis of U343 cells induced by FMU1.5 and TRAIL. Conclusion: The successful construction, expression, and purification of the extracellular domain of DR5 protein lays a foundation for further study of DR5 function.

2

Cite

Share

Effect of telomerase-antisense DNA on apoptosis of hepatoma cells induced by arsenic trioxide

Zhihua DENG ; Ziyan HAN ; Qi WANG ; Shuisheng SHI

Chinese Journal of Cancer Biotherapy.2006;0(06):-.

Objective: To observe the effect of telomerase-antisense DNA on apoptosis of hepatoma cells induced by arsenic trioxide, in an effort to look for a new anti-hepatic cancer agent with high efficiency, low cytotoxicity. Methods: We designed and synthesized a 20nt telomerase-antisense DNA targeting telomerase template and observed its influence on the telomerase activity of hepatoma cells. H-E staining, flow cytometry, and DNA agarose electrophoresis were used to study the preventive effect of telomerase-antisense DNA on hepatoma cells apoptosis induced by arsenic trioxide. Flow cytometry was used to detect the expression of Fas, Fas-L, and bcl-2. Results: Telomerase-antisense DNA (5 ?mol/L) effectively inhibited the telomerase activity of hepatoma cells after 24 hours (P

3

Cite

Share

Efficacy of 3LL/GM-CSF tumor vaccine combined with pacilitaxel chemotherapy in treatment of mice bearing transplanted Lewis lung carcinoma

Xiujuan ZHENG ; Lei ZHANG ; Yi LIN ; Yi ZHANG ; Hongming HU ; Yiwei CHU

Chinese Journal of Cancer Biotherapy.2006;0(06):-.

Objective: To evaluate the efficacy of 3LL/GM-CSF tumor vaccine combined with pacilitaxel chemotherapy in treatment of mice bearing transplanted Lewis lung carcinoma. Methods: The tumor vaccine 3LL/GM-CSF was prepared by infecting Lewis lung carcinoma cell line 3LL with adenovirus encoding GM-GSF. Mice model of Lewis lung carcinoma was established by subcutaneous injection of 2?104 3LL cells into C57BL/6(H-2b)mice. The sensitivity of Lewis lung carcinoma cell line-3LL to the treatment of pacilitaxel was detected in vivo and in vitro. The mice tumor model was first treated with pacilitaxel chemotherapy and then with 3LL/GM-CSF, or first with 3LL/GM-CSF and then with pacilitaxel. Tumor growth and the long-term survival of mice were observed after treatment. The immune memory and cytotoxicity against target cells were studied in the mice. Results: Pacilitaxel at 100 nmol/L killed 32.10% 3LL cells after 24 hour in vitro; but pacilitaxel at 5-25 mg/kg only had a poor effect on growth of 3LL cells in vivo. The tumors rebated in 70% of mice treated with pacilitaxel chemotherapy and 3LL/GM-CSF vaccination successively, and the survival of these mice was obviously longer than that of pure pacilitaxel chemotherapy group (70.0 days vs 27.5 days). The killing rate of 3LL/GM-CSF after pacilitaxel chemotherapy was 41.35% on day 3. Meanwhile, the survival mice could resist the re-attack of 3LL cells (2?104). We also noticed that first treatment with 3LL/GM-CSF and then pacilitaxel chemotherapy had no effect on tumors. Conclusion: Application of tumor vaccine shortly after pacilitaxel chemotherapy can induce specific immune responses and prolong the survival of experimental mice, which provide a basis for future clinical practice.

4

Cite

Share

Expression of vascular basement membrane-derived multifunctional peptide in Pichia pastoris and identification of its bioactivity

Wenjian DAI ; Man ZHANG ; Shuping PENG ; Jiangu CAO

Chinese Journal of Cancer Biotherapy.2006;0(06):-.

Objective: To express vascular basement membrane-derived multiple peptide (VBMDMP) in Pichia pastoris and to identify its bioactivity. Methods: PCR technique was used to obtain the target fragment GST-VBMDMP, which was then cloned into pPIC9K vector. The resultant vector pPIC9K-GST-VBMDMP was transfected into Pichia pastoris cells with spheroplast and the positive recons was identified. The His + Muts transformant was shake-cultured in MGY at 30 ℃. The culture supernatant (1 ml) was collected for preservation at-70 ℃ every 24 hours while maintaining the concentration of methanol at 0.5% in the culture medium. SDS-PAGE analysis was used to detect the protein expression after 8 days and then animal and cell experiments were performed with GST-VBMDMP. Results: SDS-PAGE analysis found that protein express increased during 1-7 days and decreased after 8 days in MGY medium; GST-VBMDMP fused protein was obtained by Glutathione Sepharose 4B and it inhibited the artery endothelial cell tube structure formation in C57BL/6 mouse; GST-VBMDMP( 2, 6, 10 mg/kg)also significantly inhibited the primary cancer Lewis mouse (tumor inhibitor rates being 96.6%, 82.1%, and 61.2%, respectively)and metastatic lung tumors(tumor inhibitor rates being 96.8 %, 87.9 %, and 75.3%, respectively)(P

5

Cite

Share

Therapeutic effect of replication-deficient adenovirus carrying p16 gene on gastric cancer xenografts in nude mice

Weiguo WANG ; Changqing SU ; Jianguo SHI ; Huizhen HU ; Linfang LI ; Juming MA

Chinese Journal of Cancer Biotherapy.1994;0(01):-.

Objective:To construct a replication-deficient adenovirus carrying p16 gene and to investigate its anti-tumor activity on human gastric cancer xenografts in nude mice.Methods:p16 cDNA was amplified by PCR and was inserted into the plasmid pSuCMV,the latter was then used to recombine the replication-deficient adenovirus AdCMV-p16 in 293 cells.Human SGC-7901 gastric cancer xenograft models were established in nude mice and were divided into 3 groups:AdCMV-p16,Ad-LacZ,and control groups.Mice in AdCMV-p16 group received intratumoral injections of 2?108 pfu/100 l AdCMV-p16(injected every other day for 5 times).Mice in control group received the same volume of virus preserving solution.The tumor volumes were measured at predefined time points.The anti-tumor effect of AdCMV-p16 was observed by p16 immunochemical study and TUNEL detection of cell apoptosis.Results:The replication-deficient adenovirus expressing p16 gene evidently inhibited the growth of human gastric cancer xenografts in nude mice(P0.05),only with a inhibition rate of 4.26%.The pathological examination showed that apoptoses were the main changes in AdCMV-p16 group,and p16 gene was found in the cancer cells.Conclusion:The replication-deficient adenovirus harboring p16 gene can recover the expression of p16 in gastric cancer cells and subsequently inhibit the growth of human gastric cancer.

6

Cite

Share

Expression of somatostatin receptor subtypes and epidermal growth factor receptor in non-small cell lung cancer and their clinical significance

Xin LIU ; Huifang TIAN ; Xiuwen WANG ; Jiyuan DING

Chinese Journal of Cancer Biotherapy.1994;0(01):-.

0.05).There was a negative correlation between the EGFR expression with the expression of SSTR2A and SSTR5 in NSCLC tissues.The 3-year survival rates were 64.5% and 65.9% in patients positive of SSTR5 and SSTR2A,respectively;and were 45.2% and 22.2% for those negative of SSTR5 and SSTR2A,respectively(P

7

Cite

Share

Effects of 5-aza-2'-deoxycytidine on apoptosis of ovarian cancer cells and on expression of mismatch repair genes

Aifeng ZHANG ; Shiqian ZHANG ; Ximei WU ; Linlin ZHANG ; Lele FU

Chinese Journal of Cancer Biotherapy.1994;0(01):-.

Objective:To observe the effect of 5-aza-2'-deoxycytidine on proliferation and apoptosis of ovarian cancer cell lines(SKOV3 and 3AO)and on expression of mismatch repair(MMR)genes(hMLH1 and hMSH2)in SKOV3 and 3AO cells.Methods:Human ovarian cancer cell lines SKOV3 and 3AO were treated for 3 d with 5-aza-CdR(0.5,5,50 ?mol/L),a specific demethylation agent,and then cultured in RPMI 1640 medium for another 7 d.The cells growth was observed by MTT assay before and after 5-aza-CdR treatment and the cell apoptosis was analyzed by flow cytometry.The expression of hMLH1 and hMSH2 mRNA was examined by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR).Results:5-aza-CdR(0.5,5,50 ?mol/L)obviously inhibited the growth of SKOV3 and 3AO cells compared in a concentration dependent manner.The apoptosis rates of SKOV3 cells were(10.59?1.57)%,(17.52?1.72)%,(34.10?1.45)% after treated with 0.5,5,50 ?mol/L 5-aza-CdR,respectively;and the apoptosis rates of 3AO cells were(11.11?2.21)%,(17.24?1.11)%,and(26.53?2.00)%,respectively,which were all markedly higher than those of control group(P

8

Cite

Share

Enhancement of cancer cell radiosensitivity by adenovirus vector carrying siRNA of Survivin gene

Yan ZHANG ; Ting SUN ; Xiangrong CAO

Chinese Journal of Cancer Biotherapy.1994;0(01):-.

Objective:To construct a recombinant adenoviral vector carrying the specific siRNA of Survivin gene,and to observe its effect on the expression of Survivin gene and on the radiosensitivity of cancer cells in tumor-bearing mice.Methods:The specific siRNA of Survivin gene was designed and synthesized,and a recombinant adenovirus AdEGFP-siRNA was subsequently constructed.SMMC 7721 xenograft models were established with nude mice and were divided into the following 5 groups:siRNA+radiotherapy and siRNA groups(intratumoral injection of AdEGFP-siRNA),siRNA(-)group(injected every other day with AdEGFP-siRNA[-],2?108 pfu/100 ?l per time,total 5 times),pure radiotherapy group and blank control groups(injected with the same volume of normal saline).On day 10,12,14,and 16,the mice in siRNA+radiotherapy and pure radiotherapy groups were given 5 Gy/time radiotherapy.The tumor volumes were measured regularly.The expression of Survivin in tumor tissues was determined immunohistochemically.Results:Adenovirus AdEGFP-siRNA harboring the specific siRNA of Survivin gene and enhanced green fluorescent protein gene(EGFP)was successfully recombined.The growth of SMMC 7721 xenografts in nude mice was inhibited after injecting AdEGFP-siRNA,with the inhibition rate being 56.2%.The inhibition rate in AdEGFP-siRNA therapy + radiotherapy increased to 82.6%.Immunohistochemistry study showed that the specific siRNA markedly silenced the expression of Survivin gene in hepatocarcinoma cells.Conclusion:The specific siRNA can markedly silence Survivin gene and subsequently inhibit the growth of cancer;meanwhile,it can also increase the radiosensitivity of cancer cells so as to improve the treatment effect.

9

Cite

Share

Herceptin combined with carboplatin inhibiting cervical cancer cells and the related mechanism

Wei WANG ; Xiaoyan XIN ; Yu LIU ; Puyuan LI ; Ye DING

Chinese Journal of Cancer Biotherapy.1994;0(01):-.

Objective:To evaluate the inhibitory effect of Herceptin on HeLa,SiHa cells and to investigate the synergistic mechanism of Herceptin and Carboplatin.Methods:HeLa and SiHa cells were treated with Herceptin(at 5,10,20,40,and 80 ?g/ml),Carboplatin(at 0.5,1,2,4,and 8 ?g/ml),and Her+CBP[(10+1),(20+2),and(40+4)?g/ml] separately.The untreated cells were taken as control.SP immunohistochemical method was used to detect the protein expression of HER-2/neu and downstream Ras oncogene.MTT method was used to study the inhibition effect of Herceptin on HeLa,SiHa cells and its synergetic effect with carboplatin.FCM was used to detect the cell apoptosis and cell cycle.The mRNA expression of her-2/neu and ras were assessed by RT-PCR;the protein expression of HER-2/neu and Ras were studied by Western blotting.Results:Herceptin significantly inhibited cervical cancer cells proliferation,and there was a synergistic effect when combined with carboplatin(P

10

Cite

Share

Effect of fetal liver AFT024 cells on multidrug resistant gene 1 transfection efficiency and in vitro expansion of CD34~+ cells derived from umbilical cord blood

Hualing ZHANG ; Zeqing WEN ; Shoumin LAN ; Changzhong LI ; Jianfeng LI

Chinese Journal of Cancer Biotherapy.1994;0(01):-.

Objective:To investigate the influence of fetal liver AFT024 cells on the transfection efficiency of multidrug resistant gene 1(MDR1)and the in vitro expansion of CD34+ cells derived from umbilical cord blood.Methods:CD34+ cells were isolated from human umbilical cord blood by MACS CD34 Progenitor Cell Isolation Kit and co-cultured with AFT024 cells(AFT024 group)or cultured alone(control group)for 7 days.During the subsequent 14 days,retrovirus carrying MDR1 gene was supplemented twice a week to transfect CD34+ cells.On the 7th,14th and 21st day after culture,the number of total nucleated cells(TNC)was counted,the ratio of CD34+ cells was assayed by flow cytometry(FCM)and the number of CD34+ cells was calculated,and colony-forming cells(CFC)were counted by methylcellulose cultures.RT-PCR method was used to detect the level of MDR1 mRNA in the transfected cells.The expression and function of P-glycoprotein(P-gp)were evaluated by FCM assay and Rhodamine-123 efflux assay,respectively.The gene transfection efficiency was calculated by drug-resistant colony-forming cells assay.Results:(1)The MDR1 mRNA level in AFT024 group than that in control group.The gene transfection efficiency in AFT024 group was significantly higher than that in control group(46.0% vs 15.2%,P0.05).On the 14th day,the expansion fold of TNCs in control group was significantly higher than that in AFT024 group(P0.05).The expansion folds of CD34+ cells and CFCs in the AFT024 group were significantly higher than that of the control group(P

DISPLAY OPTIONS

Cite

Share

Cite

Share

Cite

Share

Cite

Share

Cite

Share

Cite

Share

Cite

Share

Cite

Share

Cite

Share

Cite

Share