1.Synergistic lethal effects of cetuximab combined with chemotherapy and/or radiotherapy in laryngeal squamous carcinoma cells.
Rui HAN ; Hui HUANGFU ; Wei GAO ; Chunming ZHANG ; Na WANG ; Zhuo LI ; Binquan WANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2013;27(24):1375-1380
OBJECTIVE:
To determine the sensitivity of cetuximab induced apoptosis in laryngeal squamous carcinoma cells Hep-2, and to evaluate the synergistic killing effects and regulation mechanism of cetuximab alone or cetuximab in combination with chemotherapeutic agents (cisplatin) or radiation means on Hep-2 cells.
METHOD:
To investigate the cytotoxicities of cetuximab, cisplatin and radiation, cell counting kit-8 (CCK-8) assay was used for the detection of cell growth inhibition ratio, and fluorescence activated cell sorter FACS for the apoptotic rate and cell cycle distribution.
RESULT:
Cetuximab had inhibitive effect on Hep-2 cells within a certain range of concentration in a time- and dose-dependence manner. The inhibition concentration 50% (IC50) of cetuximab on Hep-2 cells for 24 h was 1 036.84 microg/ml. For application of cisplatin and radiation, the apoptotic rate of Hep-2 cell was higher by combining with cetuximab than their single or combined administration. Moreover, the cell cycle arrested at G0/G1 phase.
CONCLUSION
Laryngeal cancer Hep-2 cells was sensitive to the cetuximab induced apoptosis. Cetuximab combined with cisplatin and/or radiation can increase the antiproliferative effects on Hep-2 cells. These findings suggest the synergistic combination of cetuximab and cytotoxic agents was sequence depended.
Antibodies, Monoclonal, Humanized
;
pharmacology
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Apoptosis
;
drug effects
;
radiation effects
;
Cell Line, Tumor
;
Cetuximab
;
Cisplatin
;
pharmacology
;
Combined Modality Therapy
;
Humans
2.188Re-labeled herceptin inhibits proliferation of breast cancer cell line SKBR-3 in vitro.
Gui-ping LI ; Yi-fan ZHANG ; Yong-xian WANG
Journal of Southern Medical University 2006;26(10):1455-1457
OBJECTIVETo investigate the inhibitory effects of (188)Re-labeled herceptin on the proliferation in vitro of breast carcinoma cell line (SKBR-3) overexpressing HER-2/neu proto-oncogene.
METHODSHerceptin was radiolabeled with (188)Re through a direct labeling method. SKBR-3 cells were cultured with (188)Re-Herceptin at different radioactivity doses (3.7x10(4), 18.5x10(4), 37x10(4), 55.5x10(4) and 74x10(4) Bq/ml) or with (188)Re-nmIgG and (188)ReO(4)(-) for comparison. The cell proliferation inhibition was determined with MTT colorimetric assay.
RESULTS(188)Re-Herceptin could markedly inhibit the growth of SKBR-3 cells in a radioactivity dose-dependent fashion, while the effect of (188)Re-nmIgG and (188)ReO(4)(-) showed rather poor inhibitory effect in vitro. The 50% inhibition doses (IC(50)) of (188)Re-Herceptin, (188)Re-nmIgG and (188)ReO(4)(-) were 76.1x10(4) Bq/L, 139.2x10(4) Bq/L and 175x10(4) Bq/L, respectively.
CONCLUSION(188)Re-Herceptin can effectively inhibit the growth of in vitro cultured breast cancer cells overexpressing HER-2/neu, and shows much potential for clinical use in beast cancer radioimmunotherapy.
Antibodies, Monoclonal ; chemistry ; pharmacology ; Antibodies, Monoclonal, Humanized ; Antineoplastic Agents ; pharmacology ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Immunotoxins ; pharmacology ; Radioisotopes ; chemistry ; pharmacology ; Receptor, ErbB-2 ; biosynthesis ; Rhenium ; chemistry ; pharmacology ; Trastuzumab
3.Killing effect of sequential Herceptin and adriamycin treatment on breast cancer cell line in vitro.
Ke TAN ; Yi-xiang FAN ; Jing-xia MIAO ; Cheng-wei LÜ ; Xiao YAN ; Rong-cheng LUO
Journal of Southern Medical University 2006;26(2):234-236
OBJECTIVETo observe the killing effect of Herceptin and adriamycin sequentially applied on breast cancer cell line in vitro.
METHODSBT-474 human breast cancer cells in exponential growth phase were treated with Herceptin alone, adriamycin alone and their sequential administration (Herceptin before adriamycin and vice versa), respectively. Under optical microscope, the morphological changes of the cells were observed before and after drug administration. The expression rate and mean fluorescence intensity (MFI) of HER-2/neu and cell death rate were detected by flow cytometry.
RESULTSMicroscopically, the cells treated with different protocols all exhibited such changes as darkening and increase of cellular debris with irregular cell morphology. Flow cytometry revealed no significant difference in the expression rate of HER-2/neu in each group before and after treatment, but the MFI of HER-2/neu and death rate of the treated cells were significant different from those of the control group (P<0.05). The cell death rate of Herceptin-pretreated cells was significantly higher than that of adriamycin-pretreated ones (P<0.05).
CONCLUSIONHerceptin pretreatment enhances the killing effect of adriamycin on breast cancer cell line BT-474, which provides experimental evidence for designing clinical sequential biochemotherapy of breast cancer.
Antibiotics, Antineoplastic ; pharmacology ; Antibodies, Monoclonal ; pharmacology ; Antibodies, Monoclonal, Humanized ; Antineoplastic Agents ; pharmacology ; Breast Neoplasms ; metabolism ; pathology ; Cell Death ; drug effects ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Synergism ; Female ; Flow Cytometry ; Humans ; Receptor, ErbB-2 ; biosynthesis ; Trastuzumab
4.Role of epidermal growth factor receptor expression level in cetuximab cytotoxicity and antibody-dependent cell-mediated cytotoxicity effect against A549 lung cancer cell line.
Jin-yu LI ; Shun-chang JIAO ; Guo-qing ZHANG ; Sheng-jie SUN
Acta Academiae Medicinae Sinicae 2014;36(2):164-167
OBJECTIVETo investigate the role of epidermal growth factor receptor (EGFR) expression level in cetuximab cytotoxicity and antibody-dependent cell-mediated cytotoxicity (ADCC) effect against A549 lung cancer cell line.
METHODSA549 cell line and NKTm cells were used as the target cell and the effector cell, respectively. pEGFR-EGFP plasmids were transfected into A549 cells by nucleofector method. EGFR expression levels were measured by immunohistochemistry. The ADCC activity induced by cetuximab was assessed by cell counting kit-8 assay.
RESULTSA549 cells transfected with pEGFR-EGFP plasmids expressed higher level of EGFR protein on membrane and were more sensitive to ADCC activity mediated by cetuximab (P<0.05). The inhibition rate of A549 cells showed no significant difference between transfection group and wild-type group when treated with cetuximab alone (P> 0.05).
CONCLUSIONEGFR expression level influences the sensitivity of A549 lung cancer cell line to ADCC activity mediated by cetuximab but not to cetuximab alone.
Adenocarcinoma ; pathology ; Antibodies, Monoclonal, Humanized ; pharmacology ; Antineoplastic Agents ; pharmacology ; Cetuximab ; Humans ; Immunohistochemistry ; Lung Neoplasms ; pathology ; Receptor, Epidermal Growth Factor ; metabolism ; Tumor Cells, Cultured
5.The influence and safety of denosumab on bone mineral density of lumbar spine in women with low bone mass: a meta-analysis.
Journal of Southern Medical University 2013;33(6):913-917
OBJECTIVETo evaluate the influence and safety of denosumab on bone mineral density (BMD) of lumbar spine in women with low bone mass.
METHODSThe clinical literatures concerning denosumab for the treatment of osteopenia or osteoporosis in women were searched from Medline, Embase, Cochrane Central Register of Controlled Trials, Wanfang database, China National Knowledge Infrastructure database, Chinese Biomedical Database. Randomized controlled trials (RCT) were selected by the inclusive and exclusive criteria. The jadad scale was used in the quality assessment of included studies. Meta-analysis of valid data picked from included studies was performed by RevMan 5.0.24 software.
RESULTS5 RCT were included in this meta-analysis. The results of meta-analysis using the fixed effects model showed that, the increase level of lumbar BMD after 12 month was 5.45% (95% CI, 5.05%~5.84%) higher in denosumab group than in placebo control group (P<0.00001). The serious adverse event, serious infection event and pack pain occurred during the followed-up were analysed using fixed effects model. The results showed no significant difference between two groups.
CONCLUSIONCompared with placebo control group, denosumab can significant increase the BMD of lumbar spine, and the safety of two groups is similar.
Antibodies, Monoclonal, Humanized ; adverse effects ; pharmacology ; Bone Density ; drug effects ; Bone Density Conservation Agents ; adverse effects ; pharmacology ; Denosumab ; Female ; Humans ; Lumbar Vertebrae ; Randomized Controlled Trials as Topic
6.Inadvertent use of bevacizumab to treat choroidal neovascularisation during pregnancy: a case report.
Ziqiang WU ; Jingjing HUANG ; Srinivas SADDA
Annals of the Academy of Medicine, Singapore 2010;39(2):143-145
INTRODUCTIONThis study reports a case of bevacizumab administered to treat choroidal neovascularisation in a woman later discovered to be pregnant.
CLINICAL PICTUREA 25-year-old pregnant woman developed myopic choroidal neovascularisation in both eyes.
TREATMENTBoth eyes were treated with a total of 3 intravitreal injections of bevacizumab sequentially.
OUTCOMEVision improved significantly in both eyes. There were no evident pregnancy-related complications at 1 year postpartum.
CONCLUSIONAlthough anti-vascular endothelial growth factor (VEGF) therapy did not result in any detectable short-term adverse event in this mother and baby, the potential toxicity of these agents must be carefully considered in pregnant patients.
Adult ; Angiogenesis Inhibitors ; administration & dosage ; pharmacology ; therapeutic use ; Antibodies, Monoclonal ; administration & dosage ; pharmacology ; therapeutic use ; Antibodies, Monoclonal, Humanized ; Bevacizumab ; Choroidal Neovascularization ; drug therapy ; Female ; Humans ; Pregnancy ; Pregnancy Complications ; drug therapy ; Treatment Outcome
7.Effect of trastuzumab on tumor cell lines shedding high or low level of HER-2 ECD.
Cai-Yun LIU ; Wei YANG ; Jin-Feng LI ; Su-Lian SUN ; Cheng-Chao SHOU
Chinese Journal of Oncology 2007;29(2):101-105
OBJECTIVETo examine the effect of trastuzumab on cell proliferation, colony formation and changes of HER-2 proteins in human breast cancer cell line SKBR3 and human ovarian cancer cell line SKOV3 cells which overexpress p185 HER-2 but shed high or low HER-2 extracellular domain (ECD) levels.
METHODSSKBR3 cells and SKOV3 cells were treated with or without trastuzumab. Cell number and the rate of colony formation were calculated. Western blot analysis was used to detect p185 HER-2, HER-2 ECD and phospho-HER-2. Two-site ELISA assay was used for the detection of HER-2 ECD.
RESULTSTrastuzumab inhibited cell proliferation, colony formation, and decreased or eliminated the levels of two uncharacterized phospho-proteins (molar weight about 90 000 and 40 000) in SKBR3 cells shedding high level of HER-2 ECD expression. These responses were not observed in SKOV3 cells shedding low level of HER-2 ECD expression. But total p185, phospho-p185 and phospho-p95 proteins did not appear to change in SKBR3 and SKOV3 cells after treatment with trastuzumab. Trastuzumab reacts not only with proteolytic cleavage HER-2 ECD containing HER-2 ECD I , II , III and IV subdomains of p185 HER-2 extracellular domain, but also with the secreted autoinhibitor p68/ECD III a specifying 340 residues, identical to subdomains I and II from the extracellular domain of p185 HER-2, followed by a unique C-terminal sequence of 79 aa encoded by intron 8, which suggested that there may be a trastuzumab binding site on p68/ECD III a protein. Comparing with HER-2 ECD levels of the same number of SKBR3 cells, there was no significant decrease of HER-2 ECD shedding level after treatment with or without trastuzumab for 4 days in serum-free medium.
CONCLUSIONAntitumor effects of trastuzumab may be related to the two uncharacterized phospho-p90 and/or phospho-p40 proteins. There is probably a trastuzumab epitope on p68/ECD III a. The decrease of HER-2 ECD levels may be positively correlated with the number of SKBR3 cells.
Antibodies, Monoclonal ; pharmacology ; Antibodies, Monoclonal, Humanized ; Antineoplastic Agents ; pharmacology ; Blotting, Western ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Ovarian Neoplasms ; metabolism ; pathology ; Phosphorylation ; Receptor, ErbB-2 ; metabolism ; Trastuzumab
8.Effects of avastin on expression of AQP4 in Müller cells under hypoxia.
Wei CAI ; Yang CHENG ; Lina KE ; Peng ZHANG ; Guotao DENG ; Guobao LI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(4):607-612
The aim of this study was to investigate the effects of Avastin on aquaporin4 (AQP4) expression in human retinal Müller cells in vitro under hypoxia, so as to explore the mechanism of Avastin treating retinal edema. The human Müller cells were cultured using the enzymatic digestion method. Müller cells were identified under the transmission electron microscopy and by using immunofluorescence staining. By using semi-quantitative reverse transcription polymerase chain reaction (RT-PCR), the expression of AQP4 mRNA and VEGF mRNA in Müller cells cultured with 500 μmol/L CoCl(2) for 0, 3, 6, 12 and 24 h, and with 0, 100, 300, 500 and 700 μmol/L CoCl(2) for 24 h was detected. The expression of AQP4 mRNA in Müller cells cultured with 50 ng/mL exogenous vascular endothelial growth factor (VEGF) for 0, 0.5, 1, 2 and 4 h, and with 0, 25, 50 and 75 ng/mL VEGF for 24 h was detected. Amplified cDNA products of AQP4 mRNA in Müller cells cultured with 500 μmol/L CoCl(2) and 200 μg/mL Avastin for 24 h were detected. The results showed that more than 95% cells displayed positive immunofluorescence reaction. Characteristic 8-10 nm intracellular filaments could be seen in the cytoplasm under the transmission electron microscopy. In the CoCl(2) experimental groups, the expression of AQP4 mRNA and VEGF mRNA in Müller cells was increased as compared with the control group. Alteration of AQP4 mRNA and VEGF mRNA levels showed a significantly positive correlation (r (2)=0.822, P<0.05). The expression of AQP4 mRNA in Müller cells was increased by VEGF. The expression of AQP4 mRNA was significantly decreased by Avastin as compared with the control group. It is suggested that Avastin can decrease the expression of AQP4 mRNA in human Müller cells under chemical hypoxic conditions partially via VEGF path, which may be one of the mechanisms of Avastin treating retinal edema.
Antibodies, Monoclonal, Humanized
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pharmacology
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Aquaporin 4
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genetics
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metabolism
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Bevacizumab
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Cells, Cultured
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Ependymoglial Cells
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metabolism
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Gene Expression
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drug effects
;
genetics
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Humans
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Hypoxia
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genetics
;
metabolism
9.Experimental study on cetuximab and dendritic cells killing head and neck squamous cell.
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2014;28(15):1133-1136
OBJECTIVE:
This research aims to investaigate the effect of cetuximab and dendritic cells (DCs) to kill the head and neck squamous cell (HNSCC), in order to provide a new way for the patients of HNSCC.
METHOD:
DCs were induced from peripheral blood monocytes by rhIL-4, rhGM-CSF and TNF-alpha in vitro, 7days later, detecting the surface marks of DCs for example CD83, CD86, and then using MTT and flow cytometry detecting the effect T lymphocytes induced by DCs combining cetuximab to kill HNSCC; EGFR and pEGFR in each group were anlysised by Western blot.
RESULT:
It is successful to induce DCs in vitro. Mature DCs (mDCs) expressed the suface mark such as CD83, CD86 higher compared with immature DCs (imDCs). Compared with other groups, cetuximab combined with DCs significantly enhanced the cytotoxicty and apoptosis to HNSCC (P < 0.05). pEGFR were gradually reduced as the concenetration of cetuximab increasing (P < 0.05). However, comparing with the group of cetuximab, the group of cetuximab combined with DC has no significant difference at the same concentration of cetuximab. In each group EGFR also has no significant diference (P > 0.05).
CONCLUSION
Cetuximab and DCs have synergistic effects, which can significantly enhance the killing effect of HNSCC.
Antibodies, Monoclonal, Humanized
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pharmacology
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Apoptosis
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drug effects
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Carcinoma, Squamous Cell
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pathology
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Cetuximab
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Dendritic Cells
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immunology
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Head and Neck Neoplasms
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pathology
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Humans
;
Squamous Cell Carcinoma of Head and Neck
;
Tumor Cells, Cultured
10.Correlation between ras gene and the resistance of nasopharyngeal carcinoma to cetuximab.
Journal of Southern Medical University 2010;30(6):1415-1417
OBJECTIVETo explore the correlation of ras gene to the drug resistance of nasopharyngeal carcinoma to cetuximab.
METHODSCultured 5-8F/Erbitux cells were induced by stepwise exposure to increasing doses of cetuximab. MTT assay was used to determine the IC50 (half inhibitory concentration) of cetuximab and the drug resistance index (RI). Western blotting was employed to detect the protein levels of H-ras and K-ras. Real-time PCR was used to detect the expression of H-ras and K-ras. Gene sequencing was performed to identify potential mutations in H-ras and K-ras genes.
RESULTSWe successfully induced cetuximab-resistant 5-8F/Erbitux hNPC cells by stepwise exposure to increasing doses of cetuximab. After treatment with cetuximab for 3 and 5 days, the RI of 5-8F/Erbitux cells was 1.2 and 1.1, respectively. The 5-8F/Erbitux cells had increased levels of H-ras and K-ras protein expressions (P<0.001) and enhanced gene expressions of H-ras (P=0.016) and ras-p21 (P=0.113) with decreased K-ras gene expression (P=0.000). Sequence analysis identified no mutations in the H-ras and K-ras genes in codons 12, 13, 59, and 61.
CONCLUSIONGene amplification and overexpression of H-ras is the major mechanism that causes the drug resistance of 5-8F/Erbitux cells to cetuximab.
Antibodies, Monoclonal ; pharmacology ; Antibodies, Monoclonal, Humanized ; Cell Line, Tumor ; Cetuximab ; Drug Resistance, Neoplasm ; genetics ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Genes, ras ; genetics ; Humans ; Nasopharyngeal Neoplasms ; genetics ; pathology