1.Apoptosis of breast cancer cell line MCF-7 cells induced by gemcitabine and radiation.
Journal of Central South University(Medical Sciences) 2006;31(5):710-713
OBJECTIVE:
To investigate the apoptosis of human breast cell line MCF-7 cells induced by gemcitabine and radiation.
METHODS:
The MTT method was applied to study the growth inhibition of MCF-7 treated with gemcitabine, radiation, gemcitabine and radiation. The apoptosis index (AI) was analyzed by flow cytometry. The morphology of the MCF-7 cells apoptosis was observed by transmission electron microscopy.
RESULTS:
When MCF-7 cells were treated with gemcitabine at different concentrations for 24 h, the cell growth inhibition rate was increased in a concentration-dependent manner. The apoptotic indexes (AI) of MCF-7 of four groups by flow cytometry revealed. The AI of (R+D) group was significantly different from those of the radiation group and the gemcitabine group (P<0.05). Condensed chromation, nuclear fragmentation and apoptotic body of MCF-7 cells were found by transmission electron microscope.
CONCLUSION
The apoptosis of human breast cancer cell line, MCF-7 cells, could be induced by gemcitabine. Gemcitabine can significantly enhance the radiation-induced apoptosis of MCF-7 cells.
Antimetabolites, Antineoplastic
;
pharmacology
;
Apoptosis
;
drug effects
;
radiation effects
;
Breast Neoplasms
;
pathology
;
Deoxycytidine
;
analogs & derivatives
;
pharmacology
;
Female
;
Flow Cytometry
;
Humans
;
Radiation
;
Radiation-Sensitizing Agents
;
pharmacology
;
Tumor Cells, Cultured
2.Effect of wild-type p53 gene transfection on the growth and radiotherapeutic sensitivity of human glioma cells.
Wei, XIANG ; Xianli, ZHU ; Hongyang, ZHAO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2005;25(4):448-50
To evaluate the effect of wild-type p53 gene on the growth and radiotherapeutic sensitivity of human glioma cells, plasmid PC53-SN3 carrying wild-type p53 gene was transfected into U251 cells. p53 gene expression in transfected cells was detected by RT-PCR, and the cell growth inhibition and apoptosis in the absence or presence of irradiation were assessed by MTT and flow cytometry. The transfection of p53 gene into U251 cells was confirmed by RT-PCR. MTT showed that p53 gene alone induced strong inhibitory effect on the growth of U251 cells (inhibition rate (IR), (79.60 +/- 5.69)%). The killing effect of irradiation alone on U251 cells was not strong (IR: (17.06 +/- 4.35)% (17.39 +/- 1.67)% (18.73 +/- 4.68)%) and increased with the irradiation doses (3, 6, 9 Gy). When combined treatment of wild-type p53 gene transfection and irradiation was used, the effect was significantly increased (IR:(80.60 +/- 5.35)%. (90.30 +/- 1.67)%, (91.30 +/- 2.01)%). The apoptosis rate of U251 cells induced by p53 gene transfection was 17.38%. The rate induced by irradiation increased (4.61%, 4.84%, 5.40%) with the irradiation doses (3, 6, 9 Gy). The apoptosis rate was also significantly increased (17.80%, 20.03%, 22.34%) after combined treatment of p53 and irradiation with different doses (3, 6, 9 Gy). It is concluded that wild-type p53 gene and irradiation could result in synergistic inhibitory effect on the growth of human glioma cells.
Apoptosis/*radiation effects
;
Brain Neoplasms/genetics
;
Brain Neoplasms/*pathology
;
Genes, p53/*radiation effects
;
Glioma/genetics
;
Glioma/*pathology
;
Transfection
;
Tumor Cells, Cultured
3.Low-dose radiation response of the p21(WAF1/CIP1) gene promoter transduced by adeno-associated virus vector.
Mitsuru NENOI ; Kazuhiro DAINO ; Sachiko ICHIMURA ; Shin Ichiro TAKAHASH ; Teruo AKUTA
Experimental & Molecular Medicine 2006;38(5):553-564
In cancer gene therapy, restriction of antitumor transgene expression in a radiation field by use of ionizing radiation-inducible promoters is one of the promising approaches for tumor-specific gene delivery. Although tumor suppressor protein p53 is induced by low doses (<1 Gy) of radiation, there have been only a few reports indicating potential utilization of a p53-target gene promoter, such as that of the p21 gene. This is mainly because the transiently transfected promoter of p53-target genes is not much sensitive to radiation. We examined the response of the p21 gene promoter to low-dose radiation when transduced into a human breast cancer cell line MCF-7 by use of recombinant adeno-associated virus (rAAV) vectors. It was shown that the p21 gene promoter transduced by rAAV vectors was more highly radiation-responsive than that transiently transfected by electroporation. A significant induction of the p21 gene promoter by radiation of low doses down to 0.2 Gy was observed. When cells were transduced with the p21 gene promoter-driven HSVtk gene by rAAV vector, they were significantly sensitized to repetitive treatment with low dose radiation (1 Gy) in the presence of the prodrug ganciclovir. It was therefore considered that the p21 gene promoter in combination with a rAAV vector is potentially usable for the development of a low-dose radiation-inducible vector for cancer gene therapy.
X-Rays
;
Tumor Cells, Cultured
;
Transgenes/*radiation effects
;
Transduction, Genetic
;
Promoter Regions (Genetics)/*radiation effects
;
Humans
;
Genetic Vectors/*radiation effects
;
Gene Therapy/methods
;
Electroporation/methods
;
Dose-Response Relationship, Radiation
;
Cyclin-Dependent Kinase Inhibitor p21/*genetics
;
Adenoviridae
;
3' Untranslated Regions/physiology
4.The effects of gamma ray irradiation on the anti-tumor activity of peripheral blood mononuclear cells in vitro.
Xianqing ZHANG ; Daocheng WU ; Shuie MU ; Aijunn XIA ; Xiaofeng HUANG ; Xiaolan WANG
Journal of Biomedical Engineering 2009;26(1):144-152
This study sought to shed light on the killing effect of peripheral blood mononuclear cells (PBMCs) irradiated by gamma ray at a dose of 1 Gy on cultured human gastric tumor cell line MKN-28. The radiation dose rate of 17 Gy/min was used. The groups in the experiment were MKN-28 cell control group, PBMCs control group, MKN-28 tumor cells with irradiated or non-irradiated PBMCs co-cultured groups. Radiation dosage was one Gray, acridine orange/ethidium bromide (AO/EB) staining was used for observation of the killing effects of PBMCs on tumor cells in different period. Cells were harvested 240 h later and observed by transmission electron microscopy. The result showed the living period of irradiated PBMCs was shorter than that of non-irradiated PBMCs. In the irradiated and non-irradiated groups,a few PBMCs were still alive after being cultured for 240 h, but the cell volume was larger than that of lymphocytes. These cells were identified as monocytes (95%) or DCs (5%) by transmission electron microscopy. The co-culture of irradiated PBMCs and MKN-28 cells showed that tumor cells were eliminated after 96 h. As compared with the non-irradiated goup, the irradiated PBMCs had more potent ability for killing tumor. The results demonstrate that 1 Gy gamma irridiation can improve the killing effect of PBMCs on the tumor cells, and that 1 Gy gamma irritation can also induce shorter living period of lymphocytes in PBMCs cultured in vitro, but such irritation has little effect on the living period of monocytes and DCs in PBMCs.
Cell Survival
;
Coculture Techniques
;
Gamma Rays
;
Humans
;
Leukocytes, Mononuclear
;
cytology
;
immunology
;
radiation effects
;
Stomach Neoplasms
;
immunology
;
pathology
;
Tumor Cells, Cultured
5.Adaptive response to ionizing radiation induced by low dose of gamma ray in human hepatoma cell lines.
Yonsei Medical Journal 1994;35(1):77-83
When cells are exposed to a low dose of a mutagenic or clastogenic agent, they often become less sensitive to the effects of a higher dose administered subsequently. Such adaptive responses were first described in Escherichia coli. Studies on mammalian cells have been limited to human lymphocytes exposed to low doses of an alkylating agent. In this study, the adaptive response to 1 cGy of gamma rays was investigated in human tumor cells using two human hepatoma cell lines, Hep G2 and Hep 3B. Experiments were carried out by delivering 1 cGy followed by 50 cGy of gamma radiation and chromatid breaks were scored as an endpoint. The results of this study indicate that prior exposure to 1 cGy of gamma rays reduces the number of chromatid breaks induced by subsequent higher doses (50 cGy). The time necessary for the expression of the adaptive response was determined by varying the time interval between the two doses from 1 hour to 72 hours. In G2 chromatids, the adaptive response was observed both at short time intervals, as early as 1 hour, and at long time intervals. In S chromatids, however, the adaptive response was shown only at long time intervals. When 3-aminobenzamide, an inhibitor of poly (ADP-ribose) polymerase, was added after 50 cGy, adaptive responses were abolished in all the experimental groups. Therefore, it is suggested that the adaptive response can be observed in human hepatoma cell lines, which is first documented through this study.
*Adaptation, Physiological
;
Carcinoma, Hepatocellular/genetics
;
Chromosome Aberrations
;
*Gamma Ray
;
Human
;
Liver Neoplasms/genetics
;
Radiation Tolerance/*physiology
;
Support, Non-U.S. Gov't
;
Tumor Cells, Cultured/*radiation effects
6.Radiosensitivity on colorectal neoplasms by recombinant adenoviral-mediated wild-type p53 gene.
Bin-bin CUI ; Ming LIU ; Peng ZHAO ; Jia-hong ZHAO ; Xin-shu DONG
Chinese Journal of Surgery 2005;43(15):1002-1005
OBJECTIVETo study the radiosensitization on the cells of colorectal cancer transfected with recombinant adenovirus vector-mediated wild-type p53.
METHODSSW480 cells transfected by wild-type p53 were treated with 4 Gy and 6 Gy radiation. The expression of recombinant adenovirus vector-mediated wild-type p53 gene was detected by Western blotting. The inhibition rate of SW480 cells was examined by MTT, apoptotic rate by TdT-mediated dUTP nick end labeling (TUNEL) and proliferating cell nuclear antigen (PCNA) by immunohistochemical method.
RESULTSSW480 cells transfected by wild-type p53 were inhibited significantly by 4 Gy and 6 Gy radiation. The level of apoptosis increased and the expression of PCNA decreased.
CONCLUSIONCells of colorectal carcinoma transfected with wild-type p53 increases their radiation sensitivity.
Adenoviridae ; genetics ; Colorectal Neoplasms ; genetics ; pathology ; radiotherapy ; Genes, p53 ; radiation effects ; Genetic Vectors ; Humans ; Mutation ; Radiation Tolerance ; Transfection ; Tumor Cells, Cultured
7.Increase of NKG2D ligands and sensitivity to NK cell-mediated cytotoxicity of tumor cells by heat shock and ionizing radiation.
Joo Young KIM ; Young Ok SON ; Soon Won PARK ; Jae Ho BAE ; Joo Seop CHUNG ; Hyung Hoi KIM ; Byung Seon CHUNG ; Sun Hee KIM ; Chi Dug KANG
Experimental & Molecular Medicine 2006;38(5):474-484
In this study, we have investigated if current cancer therapeutic modalities including hyperthermia and ionizing radiation can increase the expression of NKG2D ligands in human cancer cell lines. The expressions of NKG2D ligands were induced by both heat shock and ionizing radiation in various cell lines including KM12, NCI-H23, HeLa and A375 cells with peaks at 2 h and 9 h after treatment, respectively, although inducibility of each NKG2D ligand was various depending on cell lines. During the induction of NKG2D ligands, heat shock protein 70 was induced by heat shock but not by ionizing radiation. These results were followed by increased susceptibilities to NK cell-mediated cytolysis after treatment with heat shock and ionizing radiation. These results suggest that heat shock and ionizing radiation induce NKG2D ligands and consequently might lead to increased NK cell-mediated cytotoxicity in various cancer cells.
Tumor Cells, Cultured
;
Receptors, Immunologic/*metabolism
;
Radiation, Ionizing
;
Neoplasms/immunology/*radiotherapy/therapy
;
*Ligands
;
Killer Cells, Natural/*immunology
;
Hyperthermia, Induced/methods
;
Humans
;
Hela Cells
;
*Heat-Shock Response/physiology
;
Heat
;
HSP70 Heat-Shock Proteins/metabolism/radiation effects
;
Gene Expression Regulation, Neoplastic/radiation effects
;
Cytotoxicity, Immunologic/*physiology/*radiation effects
;
Antigens, Surface/metabolism/radiation effects
8.Effect of anticancer drugs and desferrioxamine in combination with radiation on hepatoma cell lines.
Won Ho KIM ; Chae Yoon CHON ; Young Myung MOON ; Jin Kyung KANG ; In Suh PARK ; Heung Jai CHOI
Yonsei Medical Journal 1993;34(1):45-56
Several anticancer chemotherapeutic agents (5-fluorouracil, adriamycin and cisplatinum) and desferrioxamine, an iron chelator, were tested with regard to cytotoxicity and to the combined effect on radiation induced cell killing using two human hepatoma cell lines (HepG2 and PLC/PRF/5). Survival fractions were measured by quantitative colorimetric assay (MTT assay) and dose-response curves were plotted. MTT assay could be successfully used in the assessment of radiosensitivity in addition to chemosensitivity, because a good linear relationship between optical densities and cell numbers was observed and cells approached exponential growth for the first 7 days of culture when 5 x 10(3) or less cells were inoculated per well in our study. Steepness of the final slope (D0), width of the shoulder (D0) and the extrapolation number (n) of radiation survival curves were 1061.72 rad, 226.43 rad and 1.25 respectively in HepG2 and 1091.38 rad, 268.42 rad and 1.29 respectively in PLC/PRF/5. After combining anticancer chemotherapeutic agents and desferrioxamine with radiation, the widths of the shoulders were decreased whereas sensitizer enhancement ratios were increased as the concentration of drugs increased in both cell lines. These results suggest that neither anticancer chemotherapeutic agents nor desferrioxamine enhance cell killing induced by radiation alone, but suggested the possibility that they inhibit the repair of radiation damage.
Antineoplastic Agents/*pharmacology
;
Carcinoma, Hepatocellular/*drug therapy/*radiotherapy
;
Deferoxamine/*pharmacology
;
Human
;
Liver Neoplasms/*drug therapy/*radiotherapy
;
Support, Non-U.S. Gov't
;
Tumor Cells, Cultured/drug effects/radiation effects
9.The killing effect of focused ultrsound activating protoporphyrin IX on S180 cells.
Yaohui REN ; Xiaobing WANG ; Pan WANG ; Qiao HAO ; Quanhong LIU
Journal of Biomedical Engineering 2007;24(3):626-630
The killing effect on S180 cells was studied using the combination of protoporphyrin IX and focused ultrasound at the frequency of 2.2 MHz and different intensities. Cell viability was determined by trypan blue exclusion test, morphology changes were evaluated by means of scanning electron microscopy and transmission electron microscopy after ultrasonic exposure. The results indicated that protoporphyrin IX(PPIX) alone showed no significant effect on S180 cells when compared with that of control group. Ultrasound alone and ultrasound combined with PPIX groups showed some anti-tumor effect, which became more noticeable as the ultrasound intensity and PPIX concentration increased, and when the concentration of PPIX increased to 120 microM, the ultrasound combined with PPIX exerted a more significant anti-tumor effect than did the ultrasound alone in the same experiment.
Animals
;
Apoptosis
;
drug effects
;
radiation effects
;
Mice
;
Mice, Inbred ICR
;
Photochemotherapy
;
methods
;
Photosensitizing Agents
;
pharmacology
;
Protoporphyrins
;
pharmacology
;
Sarcoma 180
;
pathology
;
therapy
;
Sonication
;
Tumor Cells, Cultured
;
Ultrasonics
10.Effects of 50 Hz magnetic fields on gene expression in MCF-7 cells.
Guang-di CHEN ; De-qiang LU ; Huai JIANG ; Zhen-ping XU
Journal of Zhejiang University. Medical sciences 2008;37(1):15-22
OBJECTIVETo investigate whether 50 Hz magnetic fields (MF) can change the gene expression profile in MCF-7 cells and to screen MF responsive genes.
METHODSIn vitro cultured MCF-7 cells were continuously exposed or sham-exposed to 0.4 mT of 50 Hz MF for 24 hours. Affymetrix Human Genome Genechips (U133A) were applied to analyze gene expression profiles in MF exposed and sham-exposed MCF-7 cells and the data were processed with Genechip data analysis software MAS 5.0 and DMT 3.0. Real-time RT-PCR assay was employed to examine the differentially expressed genes.
RESULTThirty differentially expressed genes were screened with 100 % consistency change calls in the MF exposed MCF-7 cells. Six independent real-time RT-PCR analyses showed that SCNN1A, METTL3 and GPR137B were slightly but statistically significantly changed in MCF-7 cells after exposure to 50 Hz MF (P<0.05), while other analyzed genes exhibited slight up-and down-fluctuations in expressions and no increase or decrease in each gene expression reached statistical significance (P>0.05).
CONCLUSIONThe present study identified three 50 Hz MF responsive genes in MCF-7 cells and the biological consequences of expression changes in these MF responsive genes need to be further investigated.0.4 mT 50 Hz MF exposure for longer duration might induce DNA double-strand breaks in human lens epithelial cells in vitro.
Cell Line, Tumor ; DNA Breaks, Double-Stranded ; radiation effects ; Electromagnetic Fields ; Gene Expression ; radiation effects ; Gene Expression Profiling ; Humans ; Polymerase Chain Reaction ; Radio Waves ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Cells, Cultured