中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2012年
12期
843-846
,共4页
覃静净%曾维惠%高建武%徐磊%周莹%耿松梅
覃靜淨%曾維惠%高建武%徐磊%週瑩%耿鬆梅
담정정%증유혜%고건무%서뢰%주형%경송매
癌,鳞状细胞%纳米复合物%细胞系,肿瘤%紫外线%细胞凋亡%膜电位,线粒体
癌,鱗狀細胞%納米複閤物%細胞繫,腫瘤%紫外線%細胞凋亡%膜電位,線粒體
암,린상세포%납미복합물%세포계,종류%자외선%세포조망%막전위,선립체
Carcinoma,squamous cell%Nanocomposites%Cell line,tumor%Ultraviolet rays%Apoptosis%Membrane potential,mitochondrial
目的 探讨纳米二氧化钛(TiO2)颗粒光催化活性对人表皮鳞状细胞癌细胞株A431的生长抑制效应及机制.方法 单纯紫外线(主要波长为253.7 nm,功率30 W,光距30 cm,照射时间15 min)、不同浓度(100、200、300、400、500、600 mg/L)纳米TiO2及纳米TiO2+紫外线作用于A431细胞,采用噻唑蓝(MTT)法检测上述因素对A431细胞生长抑制率的影响；膜联蛋白V-异硫氰酸荧光素/碘化丙啶(Annexin V-FITC/PI)双染法检测A431细胞凋亡率；罗丹明123(Rho123)染色法检测A431细胞线粒体跨膜电位的变化.采用SPSS13.0统计软件进行t检验和方差分析,组间比较采用SNK检验.结果 实验干预24、48、72 h后,100、200、300、400、500、600 mg/L纳米TiO2+紫外线组A431细胞的生长抑制率增高,且呈浓度依赖效应,3个时间点不同浓度组比较,F值分别为21.54、77.56、20.27,P值均＜0.05(n=6),SNK检验示各组间差异均有统计学意义；而不同浓度单纯纳米TiO2组细胞生长抑制率在3个时间点均无明显增高,差异均无统计学意义(P值均＞ 0.05).纳米TiO2联合紫外线照射可诱导A431细胞发生凋亡,并降低A431细胞线粒体跨膜电位,100、200、400 mg/L纳米TiO2+紫外线组凋亡率分别为8.86％±0.22％、11.72％±0.29％、31.24％±0.78％,空白对照组为2.69％±0.28％,经方差分析,差异有统计学意义(F=256.61,P＜0.05,n=3)；以上3个浓度组线粒体跨膜电位总荧光强度值分别为758.48±15.42、676.60±14.35、557.71±13.12,空白对照组为2943.65±70.26,经方差分析,差异有统计学意义(F=208.57,P＜0.05,n=3),SNK检验示各组间差异均有统计学意义.结论 纳米TiO2+紫外线对A431细胞有生长抑制作用,并能诱导细胞凋亡,线粒体跨膜电位下降可能是其诱导细胞凋亡的机制之一；单纯纳米TiO2对A431细胞的生长无抑制作用.
目的 探討納米二氧化鈦(TiO2)顆粒光催化活性對人錶皮鱗狀細胞癌細胞株A431的生長抑製效應及機製.方法 單純紫外線(主要波長為253.7 nm,功率30 W,光距30 cm,照射時間15 min)、不同濃度(100、200、300、400、500、600 mg/L)納米TiO2及納米TiO2+紫外線作用于A431細胞,採用噻唑藍(MTT)法檢測上述因素對A431細胞生長抑製率的影響；膜聯蛋白V-異硫氰痠熒光素/碘化丙啶(Annexin V-FITC/PI)雙染法檢測A431細胞凋亡率；囉丹明123(Rho123)染色法檢測A431細胞線粒體跨膜電位的變化.採用SPSS13.0統計軟件進行t檢驗和方差分析,組間比較採用SNK檢驗.結果 實驗榦預24、48、72 h後,100、200、300、400、500、600 mg/L納米TiO2+紫外線組A431細胞的生長抑製率增高,且呈濃度依賴效應,3箇時間點不同濃度組比較,F值分彆為21.54、77.56、20.27,P值均＜0.05(n=6),SNK檢驗示各組間差異均有統計學意義；而不同濃度單純納米TiO2組細胞生長抑製率在3箇時間點均無明顯增高,差異均無統計學意義(P值均＞ 0.05).納米TiO2聯閤紫外線照射可誘導A431細胞髮生凋亡,併降低A431細胞線粒體跨膜電位,100、200、400 mg/L納米TiO2+紫外線組凋亡率分彆為8.86％±0.22％、11.72％±0.29％、31.24％±0.78％,空白對照組為2.69％±0.28％,經方差分析,差異有統計學意義(F=256.61,P＜0.05,n=3)；以上3箇濃度組線粒體跨膜電位總熒光彊度值分彆為758.48±15.42、676.60±14.35、557.71±13.12,空白對照組為2943.65±70.26,經方差分析,差異有統計學意義(F=208.57,P＜0.05,n=3),SNK檢驗示各組間差異均有統計學意義.結論 納米TiO2+紫外線對A431細胞有生長抑製作用,併能誘導細胞凋亡,線粒體跨膜電位下降可能是其誘導細胞凋亡的機製之一；單純納米TiO2對A431細胞的生長無抑製作用.
목적 탐토납미이양화태(TiO2)과립광최화활성대인표피린상세포암세포주A431적생장억제효응급궤제.방법 단순자외선(주요파장위253.7 nm,공솔30 W,광거30 cm,조사시간15 min)、불동농도(100、200、300、400、500、600 mg/L)납미TiO2급납미TiO2+자외선작용우A431세포,채용새서람(MTT)법검측상술인소대A431세포생장억제솔적영향；막련단백V-이류청산형광소/전화병정(Annexin V-FITC/PI)쌍염법검측A431세포조망솔；라단명123(Rho123)염색법검측A431세포선립체과막전위적변화.채용SPSS13.0통계연건진행t검험화방차분석,조간비교채용SNK검험.결과 실험간예24、48、72 h후,100、200、300、400、500、600 mg/L납미TiO2+자외선조A431세포적생장억제솔증고,차정농도의뢰효응,3개시간점불동농도조비교,F치분별위21.54、77.56、20.27,P치균＜0.05(n=6),SNK검험시각조간차이균유통계학의의；이불동농도단순납미TiO2조세포생장억제솔재3개시간점균무명현증고,차이균무통계학의의(P치균＞ 0.05).납미TiO2연합자외선조사가유도A431세포발생조망,병강저A431세포선립체과막전위,100、200、400 mg/L납미TiO2+자외선조조망솔분별위8.86％±0.22％、11.72％±0.29％、31.24％±0.78％,공백대조조위2.69％±0.28％,경방차분석,차이유통계학의의(F=256.61,P＜0.05,n=3)；이상3개농도조선립체과막전위총형광강도치분별위758.48±15.42、676.60±14.35、557.71±13.12,공백대조조위2943.65±70.26,경방차분석,차이유통계학의의(F=208.57,P＜0.05,n=3),SNK검험시각조간차이균유통계학의의.결론 납미TiO2+자외선대A431세포유생장억제작용,병능유도세포조망,선립체과막전위하강가능시기유도세포조망적궤제지일；단순납미TiO2대A431세포적생장무억제작용.
Objective To evaluate the inhibitory effect of photocatalytic titanium dioxide (TiO2)on the growth of a human epidermal squamous cell carcinoma cell line A431 and its mechanism.Methods Cultured A431 cells were classified into various groups to remain untreated (blank control group),be treated with different concentrations (100,200,300,400,500,600 mg/L) of TiO2 nanoparticles alone or in combination with ultraviolet (UV,main wavelength 253.7 nm,power 30 W,distance 30 cm,exposure duration 15 min) irradiation.After additional culture for different durations,methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate cell growth,annexin V-fluorescein isothiocyanate/propidium iodide (PI) double staining to observe cell apoptosis,and Rho123 staining to determine mitochondrial transmembrane potential.Statistical analysis was carried out using SPSS 13.0 software.Analysis of variance (AOV),t test and Student-Newman-Keuls (SNK) test were performed to assess the differences in these parameters between these groups.Results The growth of A431 cells was inhibited by pretreatment with TiO2 nanoparticles followed by UV irradiation,and the inhibitory effect was enhanced as the dose of TiO2 nanoparticles increased.As AOV and SNK test showed,there were significant differences in the growth inhibition rate among A431 cells treated with different concentrations of TiO2 nanoparticles at the three time points (24,48 and 72 hours) after UV irradiation (n =6,F =21.54,77.56,20.27,respectively,all P ＜ 0.05).No statistical inhibition was observed in the growth of A431 cells treated with TiO2 nanoparticles alone compared with untreated A431 cells (all P ＞ 0.05).Photocatalytic TiO2 nanoparticles also induced the apoptosis but decreased the mitochondrial transmembrane potential in A431 cells.In detail,the apoptosis rate was 8.86％ ± 0.22％,11.72％ ± 0.29％ and 31.24％ ± 0.78％ in A431 cells treated with TiO2nanoparticles of 100,200,400 mg/L followed by UV irradiation,respectively,compared to 2.69％ ± 0.28％ in the blank control group (n =3,F =256.61,P ＜ 0.05).Decreased mitochondrial transmembrane potential (expressed as total fluorescence intensity) was observed in A431 cells treated with TiO2 nanoparticles of 100,200,400 mg/L followed by UV irradiation compared with blank control group (758.48 ± 15.42,676.60 ± 14.35,557.71 ± 13.12vs.2943.65 ± 70.26,F =208.57,P ＜ 0.05,n =3),and SNK test also revealed statistical differences between these groups.Conclusions TiO2 nanoparticles combined with UV can inhibit the growth of but induce the apoptosis in A431 cells,which may be associated with the reduction in mitochondrial transmembrane potential in A431 cells,while TiO2 nanoparticles alone show no inhibitory effect on the growth of A431 cells.
