摘要
目的观察胆固醇对人结肠癌细胞增殖、侵袭、迁移的影响,并探讨其机制。方法将对数生长期的人结肠癌细胞株HT-29分为6组,A组不处理,B组加入胆固醇(终浓度1μmol/L)处理48 h;C、D组和E、F组采用Lipofectamine 2000分别转染阴性siRNA-Hiper Fect复合物、胆固醇酰基转移酶1(ACAT1)基因siRNA-Hiper Fect复合物,转染48 h后D、F组处理同B组。采用CFSE染色法通过流式细胞术检测细胞增殖能力,以Transwell小室观察细胞侵袭和迁移能力,利用Real time PCR法检测细胞中的ACAT1 mRNA。结果 B、D组细胞增殖率、侵袭和迁移细胞数均高于其他组(P均﹤0.05),其余组间两两比较差异无统计学意义。A、B、C、D、E、F组细胞中的ACAT1 mRNA相对表达量分别为0.96±0.02、1.48±0.05、0.94±0.02、1.44±0.02、0.54±0.36、0.63±0.05;B、D组细胞中ACAT1 mRNA相对表达量高于其他组(P均﹤0.05),E、F组细胞中ACAT1 mRNA相对表达量低于其他组(P均﹤0.05),A组与C组、B组与D组、E组与F组间两两比较差异无统计学意义。结论胆固醇能提高人结肠癌细胞的增殖、侵袭、迁移能力,其可能通过促进ACAT1基因表达发挥作用。
Objective To observe the effect of cholesterol on the proliferation, invasion and migration of human colon cancer cells and to discuss the possible mechanism. Methods The human colon cancer cells HT-29 in the logarithmic growth phase were divided into 6 groups ： group A without any treatment, group B which was added with cholesterol for 48 h （final concentration： 1 μmol/L）, group C/D and E/F which were transfected by negative siRNA -HiperFect complexes and Acyl coenzyme A cholesterol acyhransferase 1 （ ACAT1 ） gene siRNA-HiperFect complexes using Lipofectamine 2000, respectively. After 48 h, the cells in the groups D and F received the same treatment of group B. CFSE staining assay was used to evaluate the proliferation of cells. Transwell assay was used to evaluate the migration and invasion of cells. The ex- pression of ACAT1 gene was detected by real-time PCR. Results The cell proliferation rate, the cell invasion and migra- tion numbers in the group B and D were higher than those of the other groups （ all P 〈 0.05 ）, and no statistically signifi- cant difference was found between any two of groups A, C, E and F. The relative expression of ACAT1 mRNA in the groups A, B, C, D, E and F was 0.96 ±±. 02, 1.48± ±. 05,0.94 ±0.02, 1.44± 0.02, 0.54 ±0. 36 and 0.63 ± 0.05, the relative expression of ACAT1 mRNA in the groups B and D was higher than that of the other groups （ all P 〈 0. 05 ）, the relative expression of ACAT1 mRNA in the groups E and F was lower than that of the other groups （ all P 〈 0. 05 ）, and no statistically significant difference was found between group A/C, B/D and E/F. Conclusion Cholesterol can improve the proliferation, migration and invasion of human colon cancer cells by promoting ACAT1 gene expression.
出处
《山东医药》
CAS
北大核心
2017年第18期1-4,共4页
Shandong Medical Journal
基金
国家自然科学基金资助项目(81302133)
中央高校基本科研业务费专项基金资助项目(2042014kf0149)
湖北省自然科学基金资助项目(2012FKC143)
关键词
结肠癌
胆固醇
胆固醇酰基转移酶1
细胞增殖
细胞侵袭
细胞迁移
colon carcinoma
cholesterol
cholesterol acyltransferase 1
cell proliferation
cell invasion
cell migra-tion
