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目的:探讨三氧化二砷(亚砷酸,As2O3)对胰腺癌细胞增殖的抑制作用及其机制。方法:采用Cell Counting Kit-8法(CCK-8法)观察不同浓度和不同作用时间的As2O3对人类胰腺癌细胞株PC-3细胞增殖的抑制作用;采用荧光显微镜和流式细胞仪观察细胞的凋亡及生长周期的变化;采用透射电镜观察其超微结构的变化;采用免疫组织化学的方法,观察凋亡相关基因蛋白(Bcl-2和Bax)表达的变化。结果:不同剂量(0.25,0.5,1,2,4mg.L-1)的As2O3均可抑制PC-3细胞的增殖,而且抑制率呈浓度-作用时间依赖关系;荧光显微镜检测显示大量早期凋亡细胞(FITC+/PI—),流式细胞仪观察显示明显的凋亡峰出现,并使细胞周期主要被阻滞在G1期(55.19%~85.80%);电镜观察显示了As2O3作用后的PC-3细胞具有典型的凋亡细胞形态学特征:细胞膜完整、胞浆浓缩、染色质固缩、核碎裂等;免疫组化研究表明As2O3注射液对PC-3细胞表达的凋亡相关基因具有一定调节作用,即可下调Bcl-2的表达,上调Bax的表达。结论:As2O3注射液既能有效地抑制人类胰腺癌PC-3细胞株的生长,又可以诱导癌细胞凋亡,这可能与其能够调节PC-3细胞的Bcl-2和Bax等凋亡相关基因蛋白的表达有关。
Objective: To investigate the inhibitory effect of arsenic trioxide (As2O3, As2O3) on pancreatic cancer cell proliferation and its mechanism. Methods: The inhibitory effects of As2O3 on proliferation of human pancreatic cancer cell line PC-3 were observed by Cell Counting Kit-8 assay (CCK-8). The cell viability was measured by fluorescence microscopy and flow cytometry The changes of apoptosis and growth cycle were observed. The changes of ultrastructure were observed by transmission electron microscopy. The expressions of Bcl-2 and Bax protein were detected by immunohistochemistry. RESULTS: As2O3 at different dosages (0.25, 0.5, 1, 2 and 4 mg.L-1) all inhibited the proliferation of PC-3 cells in a time-dependent manner. Fluorescence microscopy revealed a large number of early apoptotic cells (FITC + / PI-). Flow cytometry showed obvious apoptotic peak appeared, and the cell cycle was mainly blocked in G1 phase (55.19% -85.80%). Electron microscopy showed that PC- 3 cells have typical morphological features of apoptotic cells: complete cell membrane, cytoplasm condensation, chromatin condensation, nuclear fragmentation, etc .; immunohistochemical studies have shown that As2O3 injection on PC-3 cells have a certain degree of apoptosis-related genes Regulatory effect, you can down-regulate Bcl-2 expression, up-regulate Bax expression. CONCLUSION: As2O3 injection can effectively inhibit the growth of human pancreatic cancer cell line PC-3 as well as induce the apoptosis of cancer cells, which may be related to its ability to regulate apoptosis-related gene proteins such as Bcl-2 and Bax in PC-3 cells Related to the expression.