Background Whether WW domain containing oxidoreductase （WWOX） gene is a tumor-suppressor is still controversial. Some researchers found that the transcription of the WWOX gene was lacking not only in tumor tissues b...Background Whether WW domain containing oxidoreductase （WWOX） gene is a tumor-suppressor is still controversial. Some researchers found that the transcription of the WWOX gene was lacking not only in tumor tissues but also in non-tumorous tissues and sometimes in normal tissues. Hence it is important to explore the role of the expression of the exogenous WWOXgene in the proliferation and apoptosis of primary cultured lung carcinoma cells. Methods Lipofection technique was used to determine primary cultured lung carcinoma cells containing the highly expressed exogenous WWOX gene and primary cultured cells with vectors as controls. An animal model of lung cancer was made by subcutaneous implantation of tumor cells into nude mice. RT-PCR, Western blotting, flow cytometry, and TUNEL were used to detect the transcription, expression of the exogenous gene and the effect of the expression of targeted genes on the proliferation and apoptosis of the primary cultured lung carcinoma cells. Results The growth, clone formation rate （CFR） （（5.33±1.53）%） of the primary lung cancer cells transfected with the WWOX gene, tumor size and weight were significantly lower than those of the non-transfected lung cancer cells （CFR： （14.33±1.53）%） and the primary tung cancer cells transfected with blank plasmids （CFR： （11.00±1.73）%, P 〈0.05）. The apoptosis level of primary lung cancer cells transfected with the WWOX gene （（40.72±5.20）%） was significantly higher than that of the non-transfected lung cancer cells （（2.76±0.02）%） and the primary lung cancer cells transfected with blank plasmids （（2.72±0.15）%, P 〈0.05）. Conclusion The expression of the exogenous WWOXgene can significantly inhibit the proliferation of lung cancer cells and induce their apoptosis, suggesting that the WWOX gene possesses tumor-suppressing effect.展开更多
The correlation between expression of Ca 2+ dependent cell adhesion molecule, cadherin, and gap junctional intercellular communication (GJIC) in human lung carcinoma PG cell line and connexin43 (Cx43) cDNA transfected...The correlation between expression of Ca 2+ dependent cell adhesion molecule, cadherin, and gap junctional intercellular communication (GJIC) in human lung carcinoma PG cell line and connexin43 (Cx43) cDNA transfected PG cell clones was investigated. Results from immunoblotting and immunofluorescent staining revealed that cultured normal human lung cells (RF) expressed N cadherin. However, the expression level of N cadherin in PG cells was very low in comparison with normal RF cells. The Cx43 transfected PG clones exhibited comparable levels of Cx43 protein, but varied in the level of N cadherin expression and in the function of GJIC as measured by scrape loading and dye transfer (SLDT) method. Positive correlation between N cadherin and GJIC was demonstrated. The in vitro and in vivo growth examination results suggest that N cadherin mediated cell cell adhesions and Cx43 functional expression, the GJIC, may work coordinately with each other in regulation of cell growth and differentiation. Deficiency in both GJIC and cell adhesion may be crucial for cell transformation.展开更多
Background： Microwave therapy is a minimal invasive procedure and has been employed in clinical practice for the treatment of various types of cancers. However, its therapeutic application in non-small-cell lung canc...Background： Microwave therapy is a minimal invasive procedure and has been employed in clinical practice for the treatment of various types of cancers. However, its therapeutic application in non-small-cell lung cancer and the underlying mechanism remains to be investigated. This study aimed to investigate its effect on Lewis lung carcinoma （LLC） tumor in vivo. Methods： Fifty LLC tumor-bearing C57BL/6 mice were adopted to assess the effect of microwave radiation on the growth and apoptosis of LLC tumor in vivo. These mice were randomly assigned to 10 groups with 5 mice in each group. Five groups were treated by single pulse microwave at different doses for different time, and the other five groups were radiated by multiple-pulse treatment of a single dose. Apoptosis of cancer cells was determined by terminal deoxynucleotidyl transferase dUTP nick-end labeling assay. Western blotting was applied to detect the expression of proteins. Results： Single pulse of microwave radiation for 5 min had little effect on the mice. Only 15-min microwave radiation at 30 mW/cm2 significantly increased the mice body temperature （2.20 ± 0.82）℃ as compared with the other groups （0.78 ± 0.29 ℃, 1.24 ± 0.52 ℃, 0.78 ± 0.42 ℃, respectively）, but it did not affect the apoptosis of LLC tumor cells significantly. Continous microwave radiation exposure, single dose microwave radiation once per day for up to seven days, inhibited cell division and induced apoptosis of LLC tumor cells in a dose- and duration-dependent manner. It upregulated the protein levels of p53, Caspase 3, Bax and downregulated Bcl-2 protein. Conclusions： Multiple exposures of LLC-bearing mice to microwave radiation effectively induced tumor cell apoptosis at least partly by upregulating proapoptotic proteins and downregulating antiapoptotic proteins. Continuous radiation at low microwave intensity Ibr a short time per day is promising in treating non-small-cell lung cancer.展开更多
目的:探讨非类固醇类抗炎药NS398对肺癌H460细胞增殖及其膜型基质金属蛋白酶-1(mem-brane type 1-matrix metalloproteinase,MT1-MMP)表达的影响。方法:应用MTT法检测细胞生长抑制率,免疫荧光法检测MT1-MMP蛋白质表达,酶联免疫吸附法(en...目的:探讨非类固醇类抗炎药NS398对肺癌H460细胞增殖及其膜型基质金属蛋白酶-1(mem-brane type 1-matrix metalloproteinase,MT1-MMP)表达的影响。方法:应用MTT法检测细胞生长抑制率,免疫荧光法检测MT1-MMP蛋白质表达,酶联免疫吸附法(enzyme-linked immunosorbentassay,ELISA)检测细胞培养液中活性基质金属蛋白酶-2(matrixmetalloproteinase-2,MMP-2)的浓度。结果:NS398可抑制H460细胞的增殖及MT1-MMP蛋白质的表达,减少H460细胞培养液中活性型MMP-2的含量,并呈剂量依赖关系。结论:NS398可能通过抑制肺癌细胞MT1-MMP表达及MMP-2的激活而抑制肺癌细胞的侵袭转移。展开更多
基金supported by a grant from The National Natural Science Foundation of China (30170914)Research Special Foundation of Higher School of Education Bureau of Liaoning (20060966,2008768,2004D162)~~
文摘Background Whether WW domain containing oxidoreductase （WWOX） gene is a tumor-suppressor is still controversial. Some researchers found that the transcription of the WWOX gene was lacking not only in tumor tissues but also in non-tumorous tissues and sometimes in normal tissues. Hence it is important to explore the role of the expression of the exogenous WWOXgene in the proliferation and apoptosis of primary cultured lung carcinoma cells. Methods Lipofection technique was used to determine primary cultured lung carcinoma cells containing the highly expressed exogenous WWOX gene and primary cultured cells with vectors as controls. An animal model of lung cancer was made by subcutaneous implantation of tumor cells into nude mice. RT-PCR, Western blotting, flow cytometry, and TUNEL were used to detect the transcription, expression of the exogenous gene and the effect of the expression of targeted genes on the proliferation and apoptosis of the primary cultured lung carcinoma cells. Results The growth, clone formation rate （CFR） （（5.33±1.53）%） of the primary lung cancer cells transfected with the WWOX gene, tumor size and weight were significantly lower than those of the non-transfected lung cancer cells （CFR： （14.33±1.53）%） and the primary tung cancer cells transfected with blank plasmids （CFR： （11.00±1.73）%, P 〈0.05）. The apoptosis level of primary lung cancer cells transfected with the WWOX gene （（40.72±5.20）%） was significantly higher than that of the non-transfected lung cancer cells （（2.76±0.02）%） and the primary lung cancer cells transfected with blank plasmids （（2.72±0.15）%, P 〈0.05）. Conclusion The expression of the exogenous WWOXgene can significantly inhibit the proliferation of lung cancer cells and induce their apoptosis, suggesting that the WWOX gene possesses tumor-suppressing effect.
文摘The correlation between expression of Ca 2+ dependent cell adhesion molecule, cadherin, and gap junctional intercellular communication (GJIC) in human lung carcinoma PG cell line and connexin43 (Cx43) cDNA transfected PG cell clones was investigated. Results from immunoblotting and immunofluorescent staining revealed that cultured normal human lung cells (RF) expressed N cadherin. However, the expression level of N cadherin in PG cells was very low in comparison with normal RF cells. The Cx43 transfected PG clones exhibited comparable levels of Cx43 protein, but varied in the level of N cadherin expression and in the function of GJIC as measured by scrape loading and dye transfer (SLDT) method. Positive correlation between N cadherin and GJIC was demonstrated. The in vitro and in vivo growth examination results suggest that N cadherin mediated cell cell adhesions and Cx43 functional expression, the GJIC, may work coordinately with each other in regulation of cell growth and differentiation. Deficiency in both GJIC and cell adhesion may be crucial for cell transformation.
文摘Background： Microwave therapy is a minimal invasive procedure and has been employed in clinical practice for the treatment of various types of cancers. However, its therapeutic application in non-small-cell lung cancer and the underlying mechanism remains to be investigated. This study aimed to investigate its effect on Lewis lung carcinoma （LLC） tumor in vivo. Methods： Fifty LLC tumor-bearing C57BL/6 mice were adopted to assess the effect of microwave radiation on the growth and apoptosis of LLC tumor in vivo. These mice were randomly assigned to 10 groups with 5 mice in each group. Five groups were treated by single pulse microwave at different doses for different time, and the other five groups were radiated by multiple-pulse treatment of a single dose. Apoptosis of cancer cells was determined by terminal deoxynucleotidyl transferase dUTP nick-end labeling assay. Western blotting was applied to detect the expression of proteins. Results： Single pulse of microwave radiation for 5 min had little effect on the mice. Only 15-min microwave radiation at 30 mW/cm2 significantly increased the mice body temperature （2.20 ± 0.82）℃ as compared with the other groups （0.78 ± 0.29 ℃, 1.24 ± 0.52 ℃, 0.78 ± 0.42 ℃, respectively）, but it did not affect the apoptosis of LLC tumor cells significantly. Continous microwave radiation exposure, single dose microwave radiation once per day for up to seven days, inhibited cell division and induced apoptosis of LLC tumor cells in a dose- and duration-dependent manner. It upregulated the protein levels of p53, Caspase 3, Bax and downregulated Bcl-2 protein. Conclusions： Multiple exposures of LLC-bearing mice to microwave radiation effectively induced tumor cell apoptosis at least partly by upregulating proapoptotic proteins and downregulating antiapoptotic proteins. Continuous radiation at low microwave intensity Ibr a short time per day is promising in treating non-small-cell lung cancer.
文摘目的:探讨非类固醇类抗炎药NS398对肺癌H460细胞增殖及其膜型基质金属蛋白酶-1(mem-brane type 1-matrix metalloproteinase,MT1-MMP)表达的影响。方法:应用MTT法检测细胞生长抑制率,免疫荧光法检测MT1-MMP蛋白质表达,酶联免疫吸附法(enzyme-linked immunosorbentassay,ELISA)检测细胞培养液中活性基质金属蛋白酶-2(matrixmetalloproteinase-2,MMP-2)的浓度。结果:NS398可抑制H460细胞的增殖及MT1-MMP蛋白质的表达,减少H460细胞培养液中活性型MMP-2的含量,并呈剂量依赖关系。结论:NS398可能通过抑制肺癌细胞MT1-MMP表达及MMP-2的激活而抑制肺癌细胞的侵袭转移。
