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目的:研究土槿皮乙酸(PAB)对人雄激素非依赖性前列腺癌(AIPC)PC-3细胞系增殖的抑制作用及其可能机制。方法:体外培养AIPC PC-3细胞,以不同浓度PAB进行处理一段时间后,再用MTT法测定PAB对PC-3细胞的生长抑制情况;用台盼蓝染色计数活细胞,用吉姆萨染色法检测凋亡细胞,进一步用Hoechst33342染色进行凋亡确认;用PI染色法检测PAB对PC-3细胞周期的影响;用考马斯亮兰G-250染色法检测PAB对细胞质蛋白的影响;用荧光双染法检测细胞微管蛋白表达。结果:PAB明显抑制体外培养的PC-3细胞生长,其抑制率在一定浓度和时间范围内呈依赖关系,作用36h的IC50值为6.20μM。PAB可以通过诱导PC-3细胞凋亡而抑制期增殖,凋亡细胞的数量随PAB浓度的增加而增加。PAB能明显增加PC-3细胞时G2期细胞的比例,G1期和S期细胞数减少,细胞周期的变化与PAB浓度相关。PAB可以减少PC-3细胞胞质内蛋白含量,抑制微管蛋白形成。结论:PAB可以抑制AIPC PC-3细胞增殖,阻滞细胞于G2/M期(M晚期),呈浓度依赖性;机制可能与PAB降低胞质蛋白量,抑制微管蛋白形成有关。
AIM: To investigate the inhibitory effect of hyaluronic acid and acetic acid (PAB) on the proliferation of human androgen-independent prostate cancer (AIPC) PC-3 cell line and its possible mechanism. Methods: AIPC PC-3 cells were cultured in vitro and treated with different concentrations of PAB for a period of time. The growth inhibition of PAB on PC-3 cells was determined by MTT assay. The viable cells were counted by Trypan blue staining, The apoptosis of PC-3 cells was detected by Hoechst33342 staining. The effect of PAB on the cell cycle of PC-3 cells was detected by PI staining. The effect of PAB on the cytoplasmic proteins was detected by Coomassie brilliant blue G-250 staining. Method to detect the expression of cell tubulin. Results: PAB significantly inhibited the growth of PC-3 cells cultured in vitro. The inhibition rate was dependent on time and concentration, and the IC50 value at 6h was 6.20μM. PAB could inhibit the proliferation of PC-3 cells by inducing apoptosis, and the number of apoptotic cells increased with the increase of PAB concentration. PAB can significantly increase the proportion of cells in G2 phase in PC-3 cells, decrease the number of cells in G1 phase and S phase, and change in cell cycle is related to the concentration of PAB. PAB can reduce the cytoplasmic protein content of PC-3 cells and inhibit tubulin formation. CONCLUSION: PAB can inhibit the proliferation of AIPC PC-3 cells in a concentration-dependent manner in G2 / M phase (M stage). The mechanism may be related to the decrease of the amount of cytoplasmic protein and the inhibition of tubulin formation by PAB.