Background: AMJ13 is a new breast cancer cell line that has been established from a 70-year-old Iraqi woman with a histological diagnosis of infiltrating ductal carcinoma. It is the first for an Iraqi population. In b...Background: AMJ13 is a new breast cancer cell line that has been established from a 70-year-old Iraqi woman with a histological diagnosis of infiltrating ductal carcinoma. It is the first for an Iraqi population. In breast cancer, angiogenesis provides the tumor tissue, which is rapidly proliferated with oxygen and nutrients, removes wastes and increases the opportunity of cancer cells to invade other organs. Methods: The AMJ13 breast cancer cell line was represented at three different passages and incubated for interval times. Microarray panel of 43 different angiogenesis markers was used to scan the supernatant for the factors. ELISA was used to quantify some of the important angiogenesis factors released in the culture medium and to confirm absence of those who was not detected by the antibody array. RT-PCR was used to confirm the gene expression (mRNA) of studied factors. Results: Microarray analysis showed that TIMP1 and two secreted at highest levels compared to the rest of the factors with low presence of endostatin. Other non-detectable factors by microarray examined by ELISA assay that showed highest expression level of VEGF-A were obtained at earliest passage, while the highest levels of FGF-b were obtained at late passage. The VEGF-D secretion was shown low concentrations at all studied passages. There is no detectable level of EGF protein in different passages and times interval tested. There are no significant differences in secretion of sICAM between different passages and incubation periods. Conclusion is that AMJ13 cell line depends on VEGF-A as main angiogenesis factor to induce micro-vessels supported by low levels of VEGF-D for lymphatic vessels formation. AMJ13 cell line depends on FGF as growth factors as in late passages it was shifted to depend mainly on FGF completely. All of this process may be regulated by TGF-β. TIMP-1 has proangiogentic effect and has feedback talk with TIMP-2. Understanding the angiogenesis process for breast cancer can give us better targets for therapy and more effective treatments.展开更多
Although the remarkable anti tumor effect of tumor necrosis factor(TNF-α)and the essential role in diverse cellular and immunological properties have been evidenced,the clinical use of TNF-αis hindered due to its to...Although the remarkable anti tumor effect of tumor necrosis factor(TNF-α)and the essential role in diverse cellular and immunological properties have been evidenced,the clinical use of TNF-αis hindered due to its toxicity.Our study was aimed to develop a new drug delivery system by binding pygelated gold nanoparticles(50 nm)with TNF-αand then investigate the anticancer activity against AMJ13 cell line.The binding of these compounds were confirmed and characterized using ultraviolet-visible spectroscopy(UV-Vis),scanning electrone microscope(SEM),and transmision electrone microscope(TEM).Varios parameters in vitro were used to examine the anticancer activity of each compound against AMJ13 cell line.Gold nanoparticles(GNPs)and TNFα-GNPs were found to exert cell growth arrest against the cancer cell line.The anti-proliferative effect of these compounds was due to cell death and inducing apoptosis as confirmed by using 4’,6-Diamidino-2-phenylindole(DAPI)fluorescent assay,flow cytometry assay,and finally mitochondrial membrane potential(MMP)staining,Real-time polymerase chanin reaction(RT-PCR)was used to detect changes in the expression of p53 protein.In addition,we studied the effect of drug delivery system on body weight on mice.In conclusion,the results of this study demonstrated that the TNFα-GNPs inhibited AMJ13 cells proliferation,resulting in apoptosis during novel pathway that involved mitochondrial damage and up-reglulated p53.Taken together,the results suggested that the TNFαloaded GNPs could be a promising therapy protocol for cancer cells and could be used for wide medical applications and offer new drug recompensing a chemotherapy drug.展开更多
文摘Background: AMJ13 is a new breast cancer cell line that has been established from a 70-year-old Iraqi woman with a histological diagnosis of infiltrating ductal carcinoma. It is the first for an Iraqi population. In breast cancer, angiogenesis provides the tumor tissue, which is rapidly proliferated with oxygen and nutrients, removes wastes and increases the opportunity of cancer cells to invade other organs. Methods: The AMJ13 breast cancer cell line was represented at three different passages and incubated for interval times. Microarray panel of 43 different angiogenesis markers was used to scan the supernatant for the factors. ELISA was used to quantify some of the important angiogenesis factors released in the culture medium and to confirm absence of those who was not detected by the antibody array. RT-PCR was used to confirm the gene expression (mRNA) of studied factors. Results: Microarray analysis showed that TIMP1 and two secreted at highest levels compared to the rest of the factors with low presence of endostatin. Other non-detectable factors by microarray examined by ELISA assay that showed highest expression level of VEGF-A were obtained at earliest passage, while the highest levels of FGF-b were obtained at late passage. The VEGF-D secretion was shown low concentrations at all studied passages. There is no detectable level of EGF protein in different passages and times interval tested. There are no significant differences in secretion of sICAM between different passages and incubation periods. Conclusion is that AMJ13 cell line depends on VEGF-A as main angiogenesis factor to induce micro-vessels supported by low levels of VEGF-D for lymphatic vessels formation. AMJ13 cell line depends on FGF as growth factors as in late passages it was shifted to depend mainly on FGF completely. All of this process may be regulated by TGF-β. TIMP-1 has proangiogentic effect and has feedback talk with TIMP-2. Understanding the angiogenesis process for breast cancer can give us better targets for therapy and more effective treatments.
文摘Although the remarkable anti tumor effect of tumor necrosis factor(TNF-α)and the essential role in diverse cellular and immunological properties have been evidenced,the clinical use of TNF-αis hindered due to its toxicity.Our study was aimed to develop a new drug delivery system by binding pygelated gold nanoparticles(50 nm)with TNF-αand then investigate the anticancer activity against AMJ13 cell line.The binding of these compounds were confirmed and characterized using ultraviolet-visible spectroscopy(UV-Vis),scanning electrone microscope(SEM),and transmision electrone microscope(TEM).Varios parameters in vitro were used to examine the anticancer activity of each compound against AMJ13 cell line.Gold nanoparticles(GNPs)and TNFα-GNPs were found to exert cell growth arrest against the cancer cell line.The anti-proliferative effect of these compounds was due to cell death and inducing apoptosis as confirmed by using 4’,6-Diamidino-2-phenylindole(DAPI)fluorescent assay,flow cytometry assay,and finally mitochondrial membrane potential(MMP)staining,Real-time polymerase chanin reaction(RT-PCR)was used to detect changes in the expression of p53 protein.In addition,we studied the effect of drug delivery system on body weight on mice.In conclusion,the results of this study demonstrated that the TNFα-GNPs inhibited AMJ13 cells proliferation,resulting in apoptosis during novel pathway that involved mitochondrial damage and up-reglulated p53.Taken together,the results suggested that the TNFαloaded GNPs could be a promising therapy protocol for cancer cells and could be used for wide medical applications and offer new drug recompensing a chemotherapy drug.
