Objective:To estimate electroporation(EP) influence on malignant and normal cells.Methods: Two cell lines including human malignant melanoma(Me-43) and normal human gingival fibroblast(HCFs) were used.EP parameters we...Objective:To estimate electroporation(EP) influence on malignant and normal cells.Methods: Two cell lines including human malignant melanoma(Me-43) and normal human gingival fibroblast(HCFs) were used.EP parameters were the following:230,1000,1 730,2 300 V/cm;30 μ s by 3 impulses for every case.The viability of cells after EP was estimated by MTT assay. The ullrastructural analysis was observed by transmission electron microscope(Zeiss EM 900). Results:In the current study we observed the intracellular effect following EP on Me-43 and HGF cells.At the conditions applied,we did not observe any significant damage of mitochondrial activity in both cell lines treated by EP.Conversely,we showed that EP in some conditions can stimulate cells to proliferation.Some changes induced by EP were only visible in electron microscopy.In fibroblast cells we observed significant changes in lower parameters of EP(230 and 1 000 V/cm).After applying higher electric field intensities(2 300 V/cm) we detected many vacuoles,myelin-like bodies and swallowed endoplasmic reticulum.In melanoma cells such strong pathological modifications after EP were not observed,in comparison with control cells. The ultrastructure of both treated cell lines was changed according to the applied parameters of EP.Conclusions:We can claim that EP conditions are cell line dependent.In terms of the intracellular morphology,human fibroblasts are more sensitive to electric field as compared with melanoma cells.Optimal conditions should be determined for each cell line.Summarizing our study,we can conclude that EP is not an invasive method for human normal and malignant cells. This technique can be safely applied in chemotherapy for delivering drugs into tumor cells.展开更多
[Objectives] The aim was to investigate the effect of cryptotanshinone on apoptosis of human melanoma A375 cells and its related mechanism of mitochondrial pathway.[Methods]The cytotoxic effect of cryptotanshinone on ...[Objectives] The aim was to investigate the effect of cryptotanshinone on apoptosis of human melanoma A375 cells and its related mechanism of mitochondrial pathway.[Methods]The cytotoxic effect of cryptotanshinone on apoptosis of human melanoma A375 cells was detected by MTT colorimetry.The apoptosis of melanoma A375 cells was detected with Annexin V-FITC/PI and observed by fluorescence inverted microscope.The expression of apoptosis-related proteins was detected by Western blotting.[Results]The viability of the A375 cells decreased with the increase of drug concentration.The fluorescence intensity of the cells increased with the treatment time.The expression of pro-apoptotic protein caspase-3 gradually increased,while the expression of apoptosis-inhibiting proteins p-AKT and Bcl-2 gradually reduced.[Conclusions]Cryptotanshinone induces apoptosis of human melanoma A375 cells via AKT signaling pathway,thus exerting a good cytotoxic effect on A375 cells.展开更多
基金Suppoted by statutory funds of Medical Wroelaw University andresearch Tellowship within"Development Program of Wroclaw Medical University"funded from European Social Fund.Human CapitalNational Cohesion Strategy(Contract No.UDA-POKL.04.01.01-00-010/08-00)
文摘Objective:To estimate electroporation(EP) influence on malignant and normal cells.Methods: Two cell lines including human malignant melanoma(Me-43) and normal human gingival fibroblast(HCFs) were used.EP parameters were the following:230,1000,1 730,2 300 V/cm;30 μ s by 3 impulses for every case.The viability of cells after EP was estimated by MTT assay. The ullrastructural analysis was observed by transmission electron microscope(Zeiss EM 900). Results:In the current study we observed the intracellular effect following EP on Me-43 and HGF cells.At the conditions applied,we did not observe any significant damage of mitochondrial activity in both cell lines treated by EP.Conversely,we showed that EP in some conditions can stimulate cells to proliferation.Some changes induced by EP were only visible in electron microscopy.In fibroblast cells we observed significant changes in lower parameters of EP(230 and 1 000 V/cm).After applying higher electric field intensities(2 300 V/cm) we detected many vacuoles,myelin-like bodies and swallowed endoplasmic reticulum.In melanoma cells such strong pathological modifications after EP were not observed,in comparison with control cells. The ultrastructure of both treated cell lines was changed according to the applied parameters of EP.Conclusions:We can claim that EP conditions are cell line dependent.In terms of the intracellular morphology,human fibroblasts are more sensitive to electric field as compared with melanoma cells.Optimal conditions should be determined for each cell line.Summarizing our study,we can conclude that EP is not an invasive method for human normal and malignant cells. This technique can be safely applied in chemotherapy for delivering drugs into tumor cells.
基金Supported by Multigrain Production and Processing Characteristic Discipline Construction ProjectPostdoctoral Scientific Research Foundation of Heilongjiang Province of China(LBH-Q13132)
文摘[Objectives] The aim was to investigate the effect of cryptotanshinone on apoptosis of human melanoma A375 cells and its related mechanism of mitochondrial pathway.[Methods]The cytotoxic effect of cryptotanshinone on apoptosis of human melanoma A375 cells was detected by MTT colorimetry.The apoptosis of melanoma A375 cells was detected with Annexin V-FITC/PI and observed by fluorescence inverted microscope.The expression of apoptosis-related proteins was detected by Western blotting.[Results]The viability of the A375 cells decreased with the increase of drug concentration.The fluorescence intensity of the cells increased with the treatment time.The expression of pro-apoptotic protein caspase-3 gradually increased,while the expression of apoptosis-inhibiting proteins p-AKT and Bcl-2 gradually reduced.[Conclusions]Cryptotanshinone induces apoptosis of human melanoma A375 cells via AKT signaling pathway,thus exerting a good cytotoxic effect on A375 cells.
