丁酸钠调控HepG2细胞的增殖、凋亡和侵袭

Sodium butyrate regulates the proliferation,apoptosis and invasion of HepG2 cells

目的研究丁酸钠对人肝癌细胞HepG2增殖和凋亡的影响,并探讨可能的作用机制。方法用不同浓度丁酸钠处理HepG2细胞后,MTT方法检测细胞的增殖能力,流式细胞技术检测细胞周期的分布和细胞凋亡,Transwell小室检测丁酸钠对HepG2细胞侵袭能力的影响。免疫荧光法检测HDAC4蛋白在HepG2细胞中的表达及定位。蛋白质印迹法(Western Blot)检测HDAC4蛋白的表达水平。结果随着丁酸钠处理浓度的增加和处理时间的延长,HepG2细胞增殖能力明显受抑制,细胞周期也发生阻滞,G1期细胞比例明显增加,而S期细胞比例明显减少。不同浓度(0,1,5,10 mmol/L)丁酸钠处理HepG2细胞24 h后,早期凋亡率分别为2.7%,4.5%,6.5%,6.7%,差异有统计学意义(F=15.1,P=0.001)。丁酸钠显著抑制细胞侵袭能力,侵袭细胞百分比分别降至72.7%(1 mmol/L)、41.7%(5 mmol/L)、21.3%(10 mmol/L),差异有统计学意义(F=202.1,P<0.001)。HDAC4蛋白在HepG2细胞中呈阳性表达,主要位于细胞质中。丁酸钠明显抑制HDAC4蛋白的表达,并呈浓度依赖性。结论丁酸钠抑制肝癌细胞系HepG2的增殖,调控细胞周期、促进凋亡,抑制细胞的侵袭能力。

Objective To determine the effect of sodium butyrate on HepG2 cells growth,apoptosis,invasion and to explore the potential mechanism.Methods HepG2 cell line was treated with different concentrations of Na Bu,and MTT assay was conducted to detect cell viability.Cell cycle distribution and apoptosis was evaluated with flow cytometry.Cell invasion was analyzed by transwell assays.Immunofluorescence staining was used to detect HDAC4 protein expression and localization in HepG2 cells.Western Blot test was performed to determine the expression of HDAC4 protein.Results With the increase of Na Bu concentration and the treatment time,the cell proliferation was significantly inhibited and the cell cycle was significantly blocked with the proportion of G1 cells increased significantly and the proportion of cells in the phase S significantly decreased.The early apoptotic rates of HepG2 cells treated with Na Bu at different concentrations(0,1,5,10 mmol/L) for 24 h were 2.7%,4.5%,6.5% and 6.7%,respectively,and the difference was statistically significant(F=15.1,P=0.001).Na Bu significantly inhibited cell invasion,and the percentage of invasive cells decreased to72.7%(1 mmol/L),41.7%(5 mmol/L) and 21.3%(10 mmol/L),respectively.The difference was statistically significant(F=202.1,P<0.001).HDAC4 protein was positively expressed in HepG2 cells and mainly located in cytoplasm.Na Bu significantly inhibited the expression of HDAC4 protein in a concentration dependent manner.Conclusion Na Bu inhibited the proliferation of HepG2 cells by regulating cell cycle,promoting apoptosis and inhibiting invasion ability.