학술논문
m iR-141表达异常对人肝癌细胞恶性生物学表型的影响 / Effect of abnormal expression of miR-141 on malignant biological behav-iors of human hepatocarcinoma cells
Document Type
Academic Journal
Author
Source
中国病理生理杂志. 32(2):215-220
Subject
Language
Chinese
ISSN
1000-4718
Abstract
目的：研究微小RNA-141（ miR-141）在人肝癌细胞和正常胎肝细胞中的表达，同时分析miR-141表达异常对人肝癌细胞恶性生物学表型的影响。方法：分别提取人肝癌细胞SMMC-7721和正常肝细胞HL-7702的总RNA，采用实时荧光定量PCR法检测miR-141的表达。采用脂质体介导的转染方法分别将miR-141 mimic转染SMMC-7721细胞，将miR-141 inhibitor转染HL-7702细胞；MTS试剂盒和BrdU-ELISA检测细胞增殖能力，流式细胞术检测转染前后细胞周期和凋亡率的变化。 Transwell实验检测miR-141表达变化对细胞体外迁移能力的影响。结果：miR-141在SMMC-7721细胞中的表达较HL-7702细胞明显下降。与空白组、脂质体组和阴性对照组相比，转染25 nmol／L miR-141 mimic的SMMC-7721细胞中，细胞增殖速度减慢，S期细胞比例降低，凋亡细胞比例上升，细胞体外迁移能力下降；转染50 nmol／L miR-141 inhibitor的HL-7702细胞中，细胞增殖速度加快，S期细胞比例上升，凋亡细胞比例下降，细胞体外迁移能力增强。结论：miR-141在人肝癌细胞中表达下降，上调miR-141表达可抑制肝癌细胞体外增殖活性和迁移能力，影响细胞周期和凋亡。在肝癌发病进程中，miR-141可能扮演抑癌基因的角色。
AIM:To investigate the expression of microRNA-141 (miR-141) in human hepatocellular carcino-ma (HCC) cell line SMMC-7721 and normal hepatocyte line HL-7702, and to analyze the effect of abnormal expression of miR-141 on the malignant biological behaviors of human hepatocarcinoma cells.METHODS:The RNA from SMMC-7721 cells and HL-7702 cells was extracted.SYBR Green real-time PCR was performed to detect the expression of miR-141. Synthetic miR-141 mimic and its negative control were transfected into the SMMC-7721 cells, and miR-141 inhibitor and its negative control were transfected into the HL-7702 cells by the method of Lipofectamine.After transfection, MTS assay and BrdU-ELISA were employed to evaluate the effect of miR-141 on the cell proliferation.Flow cytometry was used to detect cell cycle and apoptosis.The changes of migration ability were investigated by Transwell invasion assay.RESULTS:The expression of miR-141 in the SMMC-7721 cells was significantly lower than that in the HL-7702 cells ( P<0.05 ) .Com-pared with blank group, Lipofectamine group and negative control group, the proliferation of the SMMC-7721 cells trans-fected with 25 nmol/L miR-141 mimic was significantly inhibited in a time-dependent manner (P<0.05).The percenta-ges of G1 phase cells and early apoptotic rate were significantly increased when miR-141 was up-regulated, but the migra-tion ability was inhibited (P<0.05).Compared with blank group, Lipofectamine group and negative control group, the proliferation of HL-7702 cells transfected with 50 nmol/L miR-141 inhibitor was significantly increased in a time-dependent manner (P<0.05).When miR-141 was down-regulated, the percentages of G1 phase cells and early apoptotic rate were significantly decreased, but the migration ability was enhanced (P<0.05).CONCLUSION:miR-141 is down-regulated in human hepatocarcinoma cell line.Up-regulation of miR-141 will not only inhibit cell proliferation and migration ability, but also affect the cell cycle and apoptosis of SMMC-7721 cells.miR-141 may function as a tumor suppressor gene during HCC development.
AIM:To investigate the expression of microRNA-141 (miR-141) in human hepatocellular carcino-ma (HCC) cell line SMMC-7721 and normal hepatocyte line HL-7702, and to analyze the effect of abnormal expression of miR-141 on the malignant biological behaviors of human hepatocarcinoma cells.METHODS:The RNA from SMMC-7721 cells and HL-7702 cells was extracted.SYBR Green real-time PCR was performed to detect the expression of miR-141. Synthetic miR-141 mimic and its negative control were transfected into the SMMC-7721 cells, and miR-141 inhibitor and its negative control were transfected into the HL-7702 cells by the method of Lipofectamine.After transfection, MTS assay and BrdU-ELISA were employed to evaluate the effect of miR-141 on the cell proliferation.Flow cytometry was used to detect cell cycle and apoptosis.The changes of migration ability were investigated by Transwell invasion assay.RESULTS:The expression of miR-141 in the SMMC-7721 cells was significantly lower than that in the HL-7702 cells ( P<0.05 ) .Com-pared with blank group, Lipofectamine group and negative control group, the proliferation of the SMMC-7721 cells trans-fected with 25 nmol/L miR-141 mimic was significantly inhibited in a time-dependent manner (P<0.05).The percenta-ges of G1 phase cells and early apoptotic rate were significantly increased when miR-141 was up-regulated, but the migra-tion ability was inhibited (P<0.05).Compared with blank group, Lipofectamine group and negative control group, the proliferation of HL-7702 cells transfected with 50 nmol/L miR-141 inhibitor was significantly increased in a time-dependent manner (P<0.05).When miR-141 was down-regulated, the percentages of G1 phase cells and early apoptotic rate were significantly decreased, but the migration ability was enhanced (P<0.05).CONCLUSION:miR-141 is down-regulated in human hepatocarcinoma cell line.Up-regulation of miR-141 will not only inhibit cell proliferation and migration ability, but also affect the cell cycle and apoptosis of SMMC-7721 cells.miR-141 may function as a tumor suppressor gene during HCC development.