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Immunological Mornitoring After TIL Treatment of Advanced Human Malignancy

Chinese Journal of Cancer Biotherapy.1995;0(03):-.

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

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Conceptual consideration of cancer,challenges and opportunities for cancerbiotherapy

Jianren GU ; Xuetao CAO

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

The outcome of current cancer therapy is far from satisfactory despite the considerable advancements have been achievedin life science and medicine.When looking for more potent drugs or technologies for cancer therapeutics,we shouht re-evaluate ourtraditional perception of caneer and have introspection whether we have some misunderstandings of cancer biology and the relationshipbetween cancer and host.In this commentary,the authors outlined their conceptual consideration of cancer,and proposed theirviewpoints about challenges and opportunities for cancer biotherapy.

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Antigen-pulsed dendritic cell vaccine combined with hyperthermia in treatment of advanced non-small cell lung cancer

Liming ZHU ; Yiping ZHANG ; Xinmin YU ; Qinghua DENG ; Zhibing WU ; Shenglin MA

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

Objective:To evaluate the safety and clinical efficacy of antigen-pulsed dendritic cell (APDC) vaccine combined hyperthermia in treating advanced non-small cell lung cancer (NSCLC). Methods: Fourteen patients with advanced NSCLC were enrolled in this study. All patients met the selecting standard and signed informed consent. Human dendritic cells were derived from peripheral blood monocytes by co-culturing them with granulocyte macrophage-colony stimulating factor and interleukin-4. DCs vaccine was prepared from antigen pulsed immature dendritic cells in vitro. The vaccine therapy was given once every week following local hyperthermia by NRL-001 Double RF Tumor Hyperthermia system (39.5 ℃-41 ℃ for 60-120 min). Every three-week was defined as a treatment cycle. Results: All patients received 16 cycles of combined treatment. The main adverse effect included fever, chill, myalgia, transient fatigue, itching, chest distress, local rashes, and blister. Seven of 14 patients had stable condition after treatment and another seven had a progressing condition, with a clinical beneficial rate of 50%. Median time to progress was 2.7 months in the patients and the overall survival period was 2.5 to 29.3 months, with the median survival time being 4.9 months; the one year survival rate was 21.4% in our group. Conclusion: The results suggest that combination of APDC vaccine therapy and local hyperthermia is well tolerated by NSCLC patients and is clinically beneficial to the patients; the clinical value of this therapy needs to be further studied.

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Phosphorylated H2AX in predicting sensitivity of hepatic cancer cell line HepG2.215 to chemotherapeutic drugs

Lin YU ; Qing SUN

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

Objective:To study the feasibility and reliability of using phosphorylated H2AX(?H2AX)as a predictor for sensitivity of hepatic carcinoma cell HepG2.215 to chemotherapy agents: etoposide, doxorubicin, mitomycin, and cisplatin. Methods: HepG2.215 cells were exposed to etoposide, doxorubicin, mitomycin or cisplatin of 1, 2, 4 and 20 concentration index (CI). Untreated HepG2.215 cells were taken as control. The proportion of HepG2.215 cells expressing ?H2AX was measured by flow cytometry, the number of ?H2AX foci in HepG2.215 cells was measured by immunocytochemistry, and cell proliferation was measured by MTT. The correlation between the number of ?H2AX foci and the percentage of HepG2.215 cells expressing ?H2AX in HepG2.215 cells was analyzed; the correlation of CI with the percentage of HepG2.215 cells expressing ?H2AX or ?H2AX foci and inhibitory rate of cell proliferation was analyzed; and the correlation of inhibitory rate of cell praliferation with the percentage of HepG2.215 cells expressing ?H2AX or ?H2AX foci was also analyzed. Results: There was a positive correlation between the number of ?H2AX foci and the percentage of HepG2.215 cells expressing ?H2AX in HepG2.215 cells after treatment with the above 4 agents (all P

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Inhibitory effect of water-soluble preparation of curcumin on colon carcinoma C26 cell line-induced angiogenesis in mice

Jianming LI ; Heping YANG ; Zhonghong BAI ; Ruancheng XU ; Hui YAN ; Fangju WANG

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

Objective:To investigate the inhibitory effect of the water-soluble preparation of curcumin on tumor angiogenesis induced by alginate-encapsulated colon carcinoma C26 cell line in mice. Methods: The water-soluble preparation of curcumin was made by forming inclusion compound of curcumin with hydroxypropyl-?-cyclodextrin (C-HP-?-CD). Subcutaneous imbedding of alginate-encapsulated colon carcinoma C26 cells was used to create tumor bearing model in BALB/c mice. Then the mouse models were intraperitoneally injected with C-HP-?-CD, TNF-70 (as positive control), or HP-?-CD(as negative control). The angiogenesis in the alginate beads was observed under light microscope, floresencent microscope, and transmission electron microscope. The fluorescein isothiacyanate-dextran (FITC-Dextran) tracing technique was used to observe the inhibitory effect of the inclusion compound of curcumin on angiogenesis induced by colon carcinoma C26 cells in alginate beads.Results: The C-HP-?-CD and the mouse model of subcutaneous alginate-encapsulated colon carcinoma C26 cells were successfully created. Transmission electron microscope showed angiogenesis in alginate-encapsulated colon carcinoma C26 cells; claval endothelial cells were seen in some lumens. Optical microscope and the fluorescence microscope showed that the angiogenesis density of the positive group and curcumin groups were obviously lower than those of the negative control group; no obvious difference at vessel morphosis was found among the 3 alginate-encapsulated groups. FITC-Dextran tracing technology found that the fluorescent intensity of the negative control group was significantly higher than those of the positive control group and the curcumin group (P

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Fusion gene pEGFP-C3-B7.2-hTERT induces immune response in mice and restrains transplanted hepatoma

Min AI ; Shengli YANG ; Aihua JIANG

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

Objective:To construct a fusion DNA vaccine pEGFP-C3-B7.2-hTERT and to observe the capability of pEGFP-C3-B7.2-hTERT DNA vaccine to induce specific anti-tumor immune responses and to inhibit growth of the hepatoma transplanted from H22 cells. Methods: The B7.2 and hTERT cDNA (amplified by RT-PCR), together with pEGFP-C3 as the vector were used to construct fusion gene plasmid pEGFP-C3-B7.2-hTERT, which was then used to vaccinated C57BL/6 mice for 3 times at a 7 d interval. Animals vaccinated with pEGFP-C3-B7.2, pEGFP-C3-hTERT, pEGFP-C3 and PBS were taken as controls. Splenocytes CTLs of immunized mice, the levels of IL-2, IFN-? in the culture supernatant, the levels of antibodies against hTERT, and the changes of the T lymphocyte subsets in the peripheral blood were all examined. The mice harboring hepatocarcinoma H22 cells were challenged with pEGFP-C3-B7.2-hTERT and the tumor forming time and the survival periods of mice were observed. Results: Agarose gel electrophoresis, nuclease digestion and sequencing confirmed the successful construction of pEGFP-C3-B7.2-hTERT. The CTLs activity of splenocytes from the mice immunized with pEGFP-C3-B7.2-hTERT fusion gene was significantly higher than those of the other groups (P

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Inhibitory effect of antisense oligodeoxynucleotide targeting survivin on growth of human gastric cancer cells

Lantao XU ; Yu CUI

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

Objective:To investigate the inhibitory effect of antisense oligodeoxynucleotide (ASODN) targeting survivin gene on gastric cancer cell line SGC-7901. Methods: Cultured SGC-7901 cells were divided into 6 groups: control group, lipofectin group, oligodeoxynucleotide(ODN) group, 1?10-5mol/L ASODN group, 3?10-5mol/L ASODN group and 5?10-5mol/L ASODN group. After incubated with ASODN, the expression of survivin mRNA in the SGC-7901 cells was examinated by PCR; the apoptosis of SGC-7901 cells was studied by the morphological observation using inverted microscope, electron microscope, flow cytometry and DNA agarose electrophoresis; telomerase activity was determined using PCR-based TRAP (telomere repeat amplification protocol) assay; MTT assay was used to study the inhibitory effect of ASODN on the proliferation of SGC-7901 cells. Results: ASODN obviously inhibited the expression of survivin mRNA in the SGC-7901 cells. The apoptotic rate of gastric cell was significantly increased after incubated with ASODN ([7.01?0.21]% vs [25.0?0.52]%, P

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Anti-tumor effect of dendritic cell based vaccine against autologous lung cancer cells

Jun WANG ; Xiaolin PU ; Fuyin LIU ; Weifei FAN ; Lijuan MENG ; Min YANG ; Lin XU

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

Objective:To prepare dendritic cell (DC)-cytokine induced killer (CIK) tumor vaccine using DCs of patients with nonsmall-cell lung cancer (NSCLC) harboring autologous tumor antigens and to observe its inhibitory effect on autologous tumor cells in vitro. Methods: Peripheral blood mononuclear cells (PBMCs) extracted from NSCLC patients were cultured into DCs and CIKs in presence of different cytokines. Cancer cells from the patients were cultured in vitro and were used to prepare different tumor antigens: tumor cell lysates, necrotic tumor cells and normally-grown tumor cells. The DCs were stimulated by OK-432 and the phenotypes of the DCs were analyzed by flow cytometer. Then the DCs were co-cultured with CIKs. The anti-tumor effect of DC-CIK was evaluated by MTT assay. Results: Compared with the DCs harboring the antigens of necrotic tumor cells and normally-grown tumor cells, DCs pulsed with tumor cell lysates had significantly higher expression of surface molecules such as CD1a (85.1?2.7), CD80 (80.0?4.4), CD83 (75.4?5.3), and HLA-DR(80.5?7.8, all P

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Recombinant mutant human-TNF in reversing drug-resistance in ovarian cancer cell line SKOV3/DDP and the related mechanism

Suju WEI ; Haiying LIU ; Jian SHI ; Wenfeng HAN

Chinese Journal of Cancer Biotherapy.1995;0(02):-.

Objective:To investigate the reversing effect of recombinant mutant human tumor necrosis factor (rmh-TNF) on cisplatin(DDP)-resistant human ovarian cancer cell line SKOV3/DDP in vitro and the related mechanism. Methods: DDP-resistant human ovarian cancer cell line SKOV3/DDP was cultured in vitro. The cytotoxic effect of rmh-TNF to SKOV3/DDP cells was examined by MTT assay and the nontoxic dose of rmh-TNF was identified. The changes of DDP resistance was observed after cells were treated with nontoxic dose of rmh-TNF by MTT assay. The expre-ssion of GST-? protein was examined by flow cytometry at different periods after rmh-TNF intervention; RT-PCR was used to analyze the expression of MDR1gene in SKOV3/DDP cells before and after rmh-TNF treatment. Results:(1)rmh-TNF at 50-122.34 U/ml showed no evident inhibitory effect on the growth of SKOV3/DDP cells (the cell survival rate higher than 90%); and 100 U/ml was chosen for the reversing experiment ( nontoxic dose).(2)IC50 values of SKOV3/DDP cells were (23.29?0.43), (8.97?0.69) and (6.43?0.79) ?g/ml after treatment with DDP for 24, 48 and 72 h, respectively; and the values decreased to (19.50?0.50),(4.34?0.43) and (2.44?0.02)?g/ml after combined treatment with 100 U/ml rmh-TNF, respectively.(3)Expression of GST-? protein and MDR1gene decreased with the prolongation of rmh-TNF treatment. Conclusion: rmh-TNF has reversal effect on the DDP-resistant cell line SKOV3/DDP, and the mechanism may be associated with the down-regulation of GST-? protein and MDR1gene expression.

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PUMA gene transfection increases sensitivity of pancreatic cancer cell line AsPC-1 to 5-FU-induced apoptosis

Kejun ZHANG ; Dechun LI ; Dongming ZHU

Chinese Journal of Cancer Biotherapy.1995;0(02):-.

Objective:To investigate whether PUMA gene transfection can increase sensitivity of pancreatic cancer cells (PC) to 5-FU-induced apoptosis. Methods: PUMA-pCEP4 containing full length PUMA cDNA or pCEP4 was transfected into human pancreatic cancer cell line AsPC-1 by lipofectamine transfection, G418 selection was used to select positive cells. AsPC-1, AsPC-1/PUMA and AsPC-1/pCEP4 cells were separately treated with serial concentrations of 5-FU(0.01-100 ?mol/L). MTT assay was used to determine the cell survival rate in each group and IC50 of 5-FU was calculated. TUNEL,FCM and DNA ladder observation were employed to study cell apoptosis. Western blotting was performed to detect the expression of PUMA protein. Results: The 5-FU IC50 values of AsPC-1, AsPC-1/PUMA and AsPC-1/pCEP4 cells were (12?1.9)?mol/L,(1.6?0.4)?mol/L and (10.4?1.6) ?mol/L, respectively, with the sensitivity of AsPC-1/PUMA cells increased by 7.5 folds. 5-FU induced cell apoptosis of AsPC-1 cells in a dose-dependent manner, with the apoptosis of AsPC-1/PUMA cells more prominent than those of AsPC-1 and AsPC-1/pCEP4 cells. Low concentration of 5-FU (0.1 ?mol/L) induced few apoptosis of AsPC-1/pCEP4 cells([1.14?0.28]%) and AsPC-1 cells ([0.9?0.23]%), and induced apoptosis in AsPC-1/PUMA cells([6.47?1.42]%). High concentration of 5-FU (1.0 ?mol/L) induced apoptosis in all groups, with that in AsPC-1/PUMA cells([34.54?9.36]%) significantly higher than those in AsPC-1/pCEP4 cells([15.8?5.15]%) and AsPC-1 cells ([12.8?3.74]%, both P

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