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Aim To screen the most effective soyasa-ponins for reversing the drug resistance of ovarian cancer cells, and investigate the possible mechanism. Methods Cell viability assay was used to analyze the drug resistance of A2780/PTX cells treated with four different common soyasaponins respectively,in order to screen out the most effective soyasaponin. Then, the most effective soyasaponin was used to detect the ex-pression of epithelial-mesenchymal transition ( EMT)-related marker proteins, including N-cadherin and E-cadherin,with Western blot and confocal microsco-py. Finally, transwell assay and wound healing assay were applied to observe effect of soyasaponin on regula-ting cancer cell migration. Results Compared with other soyasaponins,soyasaponin Ac most effectively re-versed the drug resistance of A2780/PTX cells. The expression of N-cadherin decreased while that of E-cadherin increased in A2780/PTX cells when treated with soyasaponin Ac for 48 h. The results of transwell and wound healing assay suggested that soyasaponin Ac also reduced the migration of A2780/PTX cells. Con-clusion Soyasaponin Ac can decrease the drug resist-ance via EMT pathway and weaken the migration abili-ty of ovarian cancer cells.
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Background and purpose: Human epidermal growth factor receptor 2 (HER-2) is the member of tyrosine kinase receptor family. Its differential expression plays the key role in choosing targeted drug for breast cancer. This study focused on screening the breast cancer cell clones of different HER-2 expression levels, and studying the bi-ological characteristics of these cells. Methods: Breast cancer SK-BR-3 cells were clonally purified, and the expression level of soluble HER-2 (sHER-2) from the culture supernatant was detected by the ECLIA on ADVIA Centaur CP System. Cell clones with high expression (>50.0 ng/mL), medium expression (15.8-50.0 ng/mL) and low expression (<15.8 ng/mL) of sHER-2 were identified, respectively. This study observed the morphological changes of cell strains with differential expression levels of sHER-2 by cell culture. Besides, biological characteristics were compared by a series of experiments in vitro, such as clone formation, scratch assay, and transwell detection. Results: Compared with normal breast cells, sHER-2 was overexpressed significantly in SK-BR-3 breast cancer cells. Furthermore, the abilities of clone formation, mobility and invasion of sHER-2 high expression cell strain [(51.3±3.4)%, (50.0±0.6)% and (53.5±4.2)%] were signifi-cantly higher than those of sHER-2 medium expression [(42.0±3.7)%, (19.5±3.4)% and (33.2±3.9)%] or sHER-2 low expression [(26.7±2.9)%, (13.6±1.0)% and (28.9±5.4)%], and the differences were all statistically significant (P<0.05). Conclusion: Breast cancer cell strain with high expression level of sHER-2 can enhance cell proliferation, promote cell motility and other biological effects, which may lay the foundation for clinical screening of targeted drug therapies for breast cancer.
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Objective To evaluate the effects of malignant ascites on the morphological characteristics and the proliferation and migration abilities of the tumor cell and ovarian cancer cell lines(SKOV3)in the ovarian cancerous ascites.Methods Tumor cells extracted from the ovarian cancerous ascites were cultured in vitro with DMEM high glucose culture medium,and ovarian cancer cell lines( SKOV3) were cultured in DMEM high glucose and DMEM high glucose with different proportion of malignant ascites.The morphological characteristics of the cells were observed by optical microscope and electron microscope respectively.Cell proliferation ability was de-tected by CCK kit;The effect of SKOV3 on the migration of ovarian cancer cell lines was measured by scratch test.Results The morphological characteristics of tumor cells and ovarian cancer cell lines( SKOV3) in ovarian cancer ascites were significantly different.The proliferation ability of tumor cells was decreased without the asci-tes.The proliferation and migration abilities of SKOV3 cultured in mixed culture medium were significantly im-proved compared with the cells cultured in high glucose medium.Conclusion The change of tumor cell morphol-ogy in ascites benefits its abilities of proliferation and migration.The malignant ascites promote the abilities of pro-liferation and migration of ovarian cancer cell line(SKOV3).
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AIM:To explore the effects of confluent endothelial progenitor cells (EPCs) derived from young and aged rats on the phenotype conversion, proliferation and migration of vascular smooth muscle cells ( SMCs) .METH-ODS:Mononuclear cells were obtained from the bone marrow of young (1~2 month old) and aged (19 to 26 month old) Sprague-Dawley rats and cultured with medium DMEM/F12 ( containing 15% fetal bovine serum, endothelial cell growth supplements (ECGs) 100 g/L, 1 ×105 units/L of penicillin and streptomycin, respectively).EPCs were characterized as double positive for DiI-Ac-LDL uptake and lectin binding.Abdominal aorta was obtained from 1 to 2 month old Sprague-Dawley rats.Vascular SMCs were cultured by tissue explant method and identified byα-SM-actin immunofluorescence.In transwell co-culture system, the confluent EPCs located in the upper chamber and SMCs were seeded on the lower cham-ber.The experiments were divided into passage 3 SMCs group (P3), passage 4 SMCs group (P4), passage 4 SMCs co-culture with EPCs derived from young rats group (P4YE) and passage 4 SMCs co-culture with EPCs derived from aged rats group (P4AE).The protein expression ofα-SM-actin and osteopontin was detected by Western blotting.[3H]-TdR incor-poration assay was used to determine the proliferation.SMC migration was analyzed by scratch wound healing assay.RE-SULTS:Compared with P3 group,α-SM-actin expression in P4 group significantly decreased and osteopontin protein ex-pression obviously increased, whereas no significant change was found in P4YE group.Compared with P4 group, confluent EPCs derived from young and aged rats both markedly increased α-SM-actin and decreased osteopontin expression in P4 SMCs.Compared with aged rat-derived EPCs, young rat-derived EPCs were more effectively to induce a delayed SMC phe-notype transition (from contractile phenotype to a synthetic phenotype), and to inhibit SMC proliferation and migration. CONCLUSION:Co-culture of confluent EPC induces a delayed vascular SMC phenotype transition and inhibits SMCs pro-liferation and migration.Young rat derived EPCs are more effective to induce a delayed vascular SMC phenotype transition and has stronger inhibitory effects on SMCs proliferation and migration compared with that derived from aged rats.
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AIM:To explore the effects of transforming growth factor-α( TGF-α) in the monoclonal formation , proliferation, migration and adhesiveness of human endothelial progenitor cells ( EPCs).METHODS: The isolated and cultured EPCs were treated with various concentrations of TGF-α(final concentrations of 1, 5, 10μg/L, respectively).At the same time, the PBS control and epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) group (10μg/L TGF-αplus 1∶1 000 EGFR-TKI) were set.The effects of TGF-αon monoclonal formation , proliferation, migration and adhesiveness of EPCs were determined by clone formation experiment , thiazolyl blue tetrazolium bromide (MTT), EdU, Transwell and adhesion assays , respectively.The expression of epithelial growth receptor (EGFR) and vascular endothelial growth factor ( VEGF) were measured by Western blotting .RESULTS:Different concentrations of TGF-αall significantly induced the monoclonal formation , proliferation, migration and adhesiveness of EPCs (P<0.01), which were inhibited by EGFR-TKI.The results of Western blotting showed that TGF-αalso induced the expression of EGFR and VEGF with a cer-tain concentration effect ( P<0.01) .CONCLUSION:By combining with EGFR induced the expression of VEGF , TGF-αsignificantly promotes the related cell function of monoclonal formation , proliferation, migration, adhesiveness in EPCs.
