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目的研究海兔素(aplysin)对人乳腺癌FCM-7细胞的抑制作用,并探讨其可能的作用机制。方法 MTT法测定海兔素对人乳腺癌FCM-7细胞增殖的影响;流式细胞术测定细胞凋亡情况、细胞周期变化和增殖细胞核抗原(PCNA)、Fas、bc1-2阳性细胞百分率及细胞基质钙离子(IE Ca2+)含量变化。结果经10、20、40、60、80 mg/L海兔素处理FCM-7细胞48h后,细胞的生长增殖明显受到抑制,呈量效依赖性,其IC25和IC50值为分别为28.3和31.9 mg/L;经10和40 mg/L海兔素处理FCM-7细胞48h,出现明显的凋亡亚二倍体峰,细胞凋亡率分别为5.08%,和33.6%,且随着海兔素剂量不断增加,G0/G1期细胞所占比例也逐渐升高,而S期细胞所占比例则呈逐渐下降趋势,其中,40 mg/L海兔素组与对照组及10 mg/L海兔素组比较,差异有统计学意义(P<0.05)。经10、20、40 mg/L海兔素作用于FCM-7细胞,其Fas蛋白表达阳性细胞百分率均明显升高,分别达到(14.61±1.23、25.65±1.57、38.65±1.89)%,与对照组比较及各药物组间比较,差异有统计学意义(P<0.05);而PCNA及Bc1-2蛋白表达则逐渐下降,其中,20、40 mg/L海兔素组Bc1-2和PCNA表达均较对照组降低,且海兔素浓度越高,Bc1-2和PCNA表达越低,经统计学处理,差异有统计学意义(P<0.05)。经10、20、40 mg/L海兔素作用于FCM-7细胞,IECa2+含量也显著升高,与对照组及海兔素各组间比较,差异有统计学意义(P<0.05)。结论海兔素可诱导乳腺癌FCM-7凋亡,其作用机制可能与aplysin阻滞FCM-7细胞周期,同时提高Fas促凋亡蛋白表达和Ca2+释放,抑制PCNA和Bc1-2等细胞增殖相关蛋白表达有关。
Objective To study the inhibitory effect of aplysin on human breast cancer cell line FCM-7 and to explore its possible mechanism. Methods MTT assay was used to determine the effect of dextromethorphan on the proliferation of human breast cancer cell line FCM-7. Flow cytometry was used to detect cell apoptosis, cell cycle changes, PCNA, Fas, bcl-2 positive cells and cells Changes of matrix calcium (IE Ca2 +) content. Results FCM-7 cells were treated with 10, 20, 40, 60 and 80 mg / L sea-rabbit for 48 h, the proliferation and proliferation were significantly inhibited and the IC50 and IC50 values were 28.3 and 31.9 mg / L. The apoptotic rates of subpopulations of apoptotic cells were 5.08% and 33.6% respectively after treated with 10 and 40 mg / L searabbit for 48h, The dose of G0 / G1 phase increased gradually, while the proportion of S phase cells decreased gradually. Among them, 40 mg / L sea-rabbit group and control group and 10 mg / L sea Rabbit group, the difference was statistically significant (P <0.05). The percentage of Fas protein positive cells in FCM-7 cells treated with 10, 20 and 40 mg / L sea-rabbit significantly increased (14.61 ± 1.23,25.65 ± 1.57,38.65 ± 1.89)%, respectively. Compared with the control (P <0.05), while the expressions of PCNA and Bcl-2 protein decreased gradually. Among them, the expressions of Bcl-2 and PCNA in 20 and 40 mg / L sea-rabbits group were significantly higher than those in control group Compared with the control group, the levels of Bcl-2 and PCNA were lower than those of the control group. The statistical difference was significant (P <0.05). After treated with 10, 20 and 40 mg / L searabbit, the content of IECa2 + was also significantly increased in FCM-7 cells. The difference was statistically significant (P <0.05) compared with the control group and the sea rabbit group. Conclusions: Hydantatin can induce the apoptosis of breast cancer cell line FCM-7, and its mechanism may be related to aplysin arresting the cell cycle of FCM-7, increasing the expression of Fas and promoting apoptosis, inhibiting the proliferation of PCNA and Bc1-2 Protein expression.