CAJ | 학술논문

目的探讨整合素靶向光动力学疗法(PDT)对人胰腺癌的体外抗癌作用.方法将胰腺癌SW1990细胞分成4组,以不加量子点、无光辐照培养的细胞作为空白对照,另设单纯光辐照组、光敏剂组、PDT组.合成量子点-精氨酸甘氨酸-天冬氨酸(RGD)整合素探针,利用激光共聚焦显微镜验证其靶向性,将其作为光敏剂进行PDT.用荧光显微镜观察各组处理后48 h胰腺癌细胞的形态学变化.MTT法、流式细胞术(FCM)法检测细胞增殖、凋亡和周期的变化.RT PCR法检测各处理组细胞髓样细胞白血病-1(Mcl-1)、蛋白激酶B(Akt)、肿瘤坏死因子相关凋亡诱导配体(TRA IL)表达.荧光探针检测各组活性氧表达.组间比较采用单因素方差分析,分析PDT组的治疗效果.结果量子点-RGD探针能有效靶向标记胰腺癌细胞.MTT法PDT组24、48、72 h时的胰腺癌细胞增殖相对抑制率均高于空白对照组,差异有统计学意义(F=73.00、85.10、126.58,P均＜0.01).FCM法结果示48 h时PDT组细胞凋亡率(17.860％±1.230％)高于其他各组(F=130.617,P＜0.01)；细胞G0/G1期(69.14％±2.63％)和S期(24.41％±2.67％)出现阻滞(P均＜0.05).PDT组增殖、凋亡相关基因Mcl-1、Akt的mRNA表达低于其他各组,而凋亡诱导配体TRAIL mRNA表达高于其他各组(F=567.456、446.817,145.238,P均＜0.05).PDT组活性氧水平高于其他各组(F=3262.559,P＜0.01).结论量子点-RGD整合素靶向探针介导的PDT能明显抑制胰腺癌细胞增殖,促进细胞凋亡.
목적 탐토정합소파향광동역학요법(PDT)대인이선암적체외항암작용.방법 장이선암SW1990세포분성4조,이불가양자점、무광복조배양적세포작위공백대조,령설단순광복조조、광민제조、PDT조.합성양자점-정안산감안산-천동안산(RGD)정합소탐침,이용격광공취초현미경험증기파향성,장기작위광민제진행PDT.용형광현미경관찰각조처리후48 h이선암세포적형태학변화.MTT법、류식세포술(FCM)법검측세포증식、조망화주기적변화.RT PCR법검측각처리조세포수양세포백혈병-1(Mcl-1)、단백격매B(Akt)、종류배사인자상관조망유도배체(TRA IL)표체.형광탐침검측각조활성양표체.조간비교채용단인소방차분석,분석PDT조적치료효과.결과 양자점-RGD탐침능유효파향표기이선암세포.MTT법PDT조24、48、72 h시적이선암세포증식상대억제솔균고우공백대조조,차이유통계학의의(F=73.00、85.10、126.58,P균＜0.01).FCM법결과시48 h시PDT조세포조망솔(17.860％±1.230％)고우기타각조(F=130.617,P＜0.01)；세포G0/G1기(69.14％±2.63％)화S기(24.41％±2.67％)출현조체(P균＜0.05).PDT조증식、조망상관기인Mcl-1、Akt적mRNA표체저우기타각조,이조망유도배체TRAIL mRNA표체고우기타각조(F=567.456、446.817,145.238,P균＜0.05).PDT조활성양수평고우기타각조(F=3262.559,P＜0.01).결론 양자점-RGD정합소파향탐침개도적PDT능명현억제이선암세포증식,촉진세포조망.
Objective To investigate the anti carcinoma role of integrin targeted photodynamic therapy (PDT) on pancreatic carcinoma cells in vitro.Methods Pancreatic carcinoma cells SW1990 were divided into four groups:cells without quantum dots (QDs) and light-treated as blank control group,pure light-treated group,photosensitizer group and PDT group.The targeting of QDs-arginine,glycine,aspartic acid (RGD) and integrin probe was confirmed by laser confocal microscopy.And as a photosensitizer for photodynamic therapy,after treated for 48 hours the morphology changes of pancreatic carcinoma cells of each group were observed.After 48 hours,the cell proliferation,apoptosis and cell cycle changes were detected by methyl thiazolyl tetrazolium (MTT) assay and flow cytometry (FCM).The expressions of myeloid cell leukemia-1 (Mcl-1),protein kinase B(Akt) and tumor necrosis factor-related apoptosis inducing ligand (TRAIL) were detected by reverse transcriptase polymerase chain reaction(RT-PCR).The amount of reactive oxygen species (ROS) of each group were evaluated by fluorescence probe.One-way ANOVA was performed for comparison between groups to analyze the treatment effects of PDT group.Results The QDs RGD probe could effectively targeting pancreatic carcinoma cells.The MTT results indicated that the relative inhibition rate of pancreatic carcinoma cells proliferation of PDT group was statistically higher than that of the other groups at 24,48,72 h (F=73.00,85.10,126.58; all P＜0.01).The FCM results revealed that the cell apoptosis rate of PDT group (17.860％ ±1.230％) was higher than that of the other groups (F=130.617,P＜0.01) and cell cycle G0/G1 phase (69.14％±2.63％) and S phase (24.41％ ± 2.67 ％) retardance was also significant (all P＜0.05).The expression of proliferation and apoptosis related gene Mcl-1 and Akt at mRNA level was lower than that of the other groups however the expression of apoptosis-inducing ligand TRAIL at mRNA level was higher than that of the other groups (F=567.456,446.817,145.238; all P＜0.05).The ROS level of PDT group was higher than that of the other groups (F=3262.559,P＜0.01).Conclusion PDT with a QDs-RGD probe could significantly inhibit pancreatic carcinoma cell proliferation and promote cell apoptosis.