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目的:研究昆布多糖硫酸酯(LAMS)对体外培养的非激素依赖型人前列腺癌细胞株PC-3的作用,以及对PC-3细胞PTEN和P27kip1基因表达的影响,并探讨其抗肿瘤的作用机制。方法:分别用0、50、100、200μg/ml浓度的LAMS作用于PC-3细胞,用WST-8法检测其对细胞生长的抑制作用,利用流式细胞术(FCM)分析细胞周期和凋亡的变化,用荧光显微镜观察PC-3细胞形态的变化,RT-PCR和Western印迹检测细胞增殖、凋亡相关基因PTEN和P27kip1mRNA及蛋白的表达。结果:LAMS能抑制PC-3细胞的增殖,呈时间-剂量效应;不同浓度LAMS诱导PC-3细胞出现剂量依赖性S期阻滞;LAMS作用于PC-3细胞后凋亡率增加,并出现凋亡的形态学改变,相关基因PTEN和P27kip1mRNA及蛋白的表达呈剂量依赖性增加。结论:LAMS能抑制体外PC-3细胞的生长,并促进其S期阻滞和凋亡,PTEN和P27kip1mRNA及蛋白的表达明显增加,可能是其抑制前列腺癌生长的机制之一。
Objective: To investigate the effect of laminarin sulfate (LAMS) on the expression of PTEN and P27kip1 in PC-3 cells and to study the antitumor effect of laminarin (PC) on the non-hormone-dependent human prostate cancer cell line PC-3 mechanism. Methods: PC-3 cells were treated with 0, 50, 100 and 200 μg / ml LAMS respectively. The inhibitory effect on cell growth was detected by WST-8 assay. Cell cycle and apoptosis were analyzed by flow cytometry The morphological changes of PC-3 cells were observed by fluorescence microscopy. The expression of PTEN and P27kip1 mRNA and protein were detected by RT-PCR and Western blot. Results: LAMS could inhibit the proliferation of PC-3 cells in a dose-time-dependent manner. L-SAMP induced a dose-dependent S-phase arrest in PC-3 cells. LAMS increased the apoptotic rate in PC-3 cells Apoptosis morphological changes, related genes PTEN and P27kip1mRNA and protein expression in a dose-dependent manner. CONCLUSION: LAMS can inhibit the growth of PC-3 cells in vitro and promote its arrest and apoptosis in S phase. The expression of PTEN and P27kip1 mRNA and protein may be one of the mechanisms that may inhibit the growth of PC-3 cells.