脱氧胆酸对正常食管细胞及食管癌细胞增殖能力的影响

Effect and mechanism of deoxycholic acid on the proliferation of normal esophageal and esophageal cancer cells

[目的]研究脱氧胆酸(DCA)对正常食管细胞及食管癌细胞增殖能力影响及机制。[方法]采用不同浓度:0μmol/L(对照组)、100μmol/L、200μmol/L的DCA分别处理永生化的正常食管细胞株Het-1 A和食管癌细胞株EC109。利用CCK8增殖实验检测5 d内不同浓度DCA对Het-1 A和EC109的增殖能力影响;提取细胞RNA,利用逆转录PCR检测不同浓度DCA对Het-1 A和EC109中细胞增殖相关基因EGFR及AKT1的表达情况变化。[结果]CCK8增殖实验结果显示:与对照组相比,200μmol/L DCA处理细胞3~5 d与对照组相比,细胞增殖能力明显受到抑制(第3天P=0.000,第4天P=0.001,第5天P=0.015),而100μmol/L DCA组细胞增殖仅在第5天时明显受到抑制(P=0.045),100μmol/L DCA处理组的细胞EC109(2~5 d:P>0.05)的增殖能力不受影响。200μmol/L DCA处理组与对照组相比,EGFR和AKT1的mRNA表达均显著降低(EGFR:P=0.039,AKT1:P=0.013);100μmol/L DCA组与对照组相比,EGFR和AKT1的mRNA表达也都显著降低(EGFR:P=0.035,AKT1:P=0.050),在Het-1 A差异无统计学意义(P=0.076)。[结论]脱氧胆酸可能通过抑制食管细胞中增殖相关基因EGFR和AKT1的表达而抑制食管细胞的增殖能力,但存在时间及浓度依赖性。

[Objective]To study the effect and mechanism of deoxycholic acid(DCA)on the proliferation of normal esophageal and esophageal cancer cells.[Methods]The immortalized normal esophageal cell line Het-1 A and esophageal cancer cell line EC109 were treated with DCA at different concentrations(0μmol/L,100μmol/L,200μmol/L).The effect of different concentrations of DCA on the proliferation of Het-1 A and EC109 in 5 days was detected by CCK8 proliferation assay.Cellular RNA was extracted and the expression of EGFR and AKT1 in Het-1 A and EC109 were detected by reverse transcription PCR.[Results]The results of CCK8 proliferation assay showed that Het-1 A(Day3-5:P<0.015)and EC109(Day 2-5:P=0.000)were treated with 200μmol/L DCA treated group compared with the control group treated with 0μmol/L DCA.The proliferative capacity was significantly inhibited.The proliferation of Het-1 A(Day 5:P<0.045)in the 100μmol/L DCA treatment group was inhibited only on the 5 th day,and the cell EC109 in the 100μmol/L DCA treatment group(Day2-5:P>0.05)was not affected.Compared with the control group treated with 0μmol/L DCA,EGFR expression was decreased in Het-1 A and EC109 cells in the 200μmol/L DCA-treated group(Het-1 A:P=0.008,EC109:P=0.039),and AKT1 expression was decreased(Het-1 A:P=0.045,EC109:P=0.013);while in the 100μmol/L DCA group,Het-1 A and EC109,EGFR expression was decreased(Het-1 A:P=0.011,EC109:P=0.035),AKT1 only The expression was significantly decreased in EC109(P=0.050)and not statistically significant in Het-1 A(P=0.076).[Conclusion]Deoxycholic acid may inhibit the proliferation of esophageal cells by inhibiting the expression of proliferation-associated genes EGFR and AKT1 in esophageal cells,but in a time-and concentration-dependent manner.