沉默酰基辅酶A-胆固醇酰基转移酶1基因对人前列腺癌细胞生物学行为的影响

摘要

目的 观察ACAT1在人前列腺癌PC-3细胞增殖和侵袭迁移中的作用.方法 体外培养人前列腺癌PC-3细胞,通过脂质体HiperFect将ACAT1 siRNA转染至人前列腺癌PC-3细胞中沉默酰基辅酶A-胆固醇酰基转移酶1(ACAT1)基因的表达,Rea1-time PCR验证沉默的效果.利用CFSE染色法分别检测加入无血清1640培养基的空白对照组、加入阴性-siRNA-HiperFect复合物的阴性对照组和加入ACAT1 siRNA-HiperFect复合物的A-CAT1组人前列腺癌PC-3细胞的增殖率,比较ACAT1基因沉默前后细胞增殖的变化.利用Transwell小室法检测ACAT1基因沉默前后人前列腺癌PC-3细胞侵袭和迁移能力的变化.结果 与空白对照组和阴性对照组比较,ACAT1 siRNA组转染48 h ACAT1 mRNA表达水平最低(P＜0.05).与阴性对照组和空白对照组比较,ACAT1siRNA组细胞增殖率下降,侵袭和迁移实验中穿膜细胞数减少,差异有高度统计学意义(P＜0.01).阴性对照组与空白对照组比较,细胞增殖率、侵袭和迁移能力无变化,差异无统计学意义(P＞0.05).结论 ACAT1基因干扰能抑制PC-3细胞的增殖,减弱侵袭迁移能力,为探寻前列腺癌发病机制和治疗提供新的思路和手段.%Objective To observe the effect of ACAT1 on human prostate cancer cells proliferation,invasion and migration.Methods Human prostate cancer cells (PC-3 cells) were cultured,then the expression of ACAT1 gene in PC-3 cells were interfered using Hiperfect transfection reagent,the expression of ACAT1 gene before and after transfection was detected by Real-time PCR.The proliferation rates of PC-3 cells in the blank control group (serum-free 1640 medium added),the negative control group (negative-siRNA-HiperFect complex added) and the ACAT1 group (ACAT1 siRNA-HiperFect complex added) were analyzed by CFSE staining.The changes of cell proliferation before and after ACAT1 gene silencing were compared.Transwell assays were used to evaluate the effect of ACAT1 gene interfering on cells migration and invasion.Results Compared with the blank control group and the negative control group respectively,ACAT1 mRNA expression level reached the lowest level after 48 hours in the ACAT1 siRNA group (P ＜ 0.05).In the ACAT1 siRNA group,compared with the blank control group and the negative control group,proliferation rate of PC-3 cell decreased,the numbers of invasive cell and migration cell reduced,the differences were statistically significant (P ＜ 0.01).Compared between the blank control group and the negative control group,the differences in proliferation rate,invasion and migration ability of human prostate cancer PC-3 cells had no statistically significant (P ＞ 0.05).Conclusion ACAT1 siRNA reduces proliferation,migration and invasion of PC-3 cells,which maybe provide a new idea and strategy for pathogenesis and treatment of prostate cancer.