摘要
目的观察微小RNA（miRNA，miR）-942在肝细胞肝癌（HCC）组织中的表达及其对肝癌细胞增殖、侵袭能力的影响。方法采用实时定量聚合酶链反应（Real-time PCR）检测70例肝癌和癌旁组织miR-942的表达；将Huh7细胞分别转染miRNA随机序列（阴性对照组）、miR-942模拟物（miR-942模拟物组）和miR-942抑制物（miR-942抑制物组）后，噻唑蓝（MTT）法和Transwell实验观察Huh7细胞增殖和侵袭能力变化；Western blot检测Huh7细胞增殖细胞核抗原（PCNA）和基质金属蛋白酶-9（MMP-9）的蛋白表达。结果miR-942在肝癌组织中表达量（0.92±0.08）与癌旁组织（0.42±0.05）比较显著上调（t＝5.712，P＝0.000）；MTT细胞增殖实验结果显示5 d后miR-942抑制物组Huh7细胞吸光度（A）值显著低于阴性对照组和miR-942模拟物组（0.79±0.02比1.02±0.04，t＝7.513，P＝0.001；0.79±0.02比1.51±0.12，t＝8.621，P＝0.001）；Transwell结果显示miR-942抑制物组侵袭转移细胞数明显减少[（59.13±5.41）个比（97.26±11.13）个，t＝11.712，P＝0.000；（59.13±5.41）个比（99.42±12.13）个，t＝12.113，P＝0.000]；Western blot结果显示PCNA和MMP-9蛋白在miR-942抑制物组的表达分别下调了80.1%和56.3%（t＝5.231，P＝0.000；t＝6.415，P＝0.000）。结论miR-942在肝癌组织中高表达，下调miR-942可抑制肝癌细胞的增殖和侵袭，其机制与下调PCNA和MMP-9表达有关。
Objective To investigate the expression of microRNA （miRNA, miR）- 942 in liver cancer tissues and its effect on proliferation and invasion of hepatocellular carcinoma （HCC）.Methods miR- 942 expression was detected in HCC and peritumoral tissues by real-time quantitative polymerase chain reaction （Real-time PCR） from 70 patients. The Huh7 cells were transfected with scrambled miRNA sequence （negative control group）, miR- 942 mimics （miR- 942 mimics group） and miR- 942 inhibitors （miR- 942 inhibitors group） respectively. The proliferative activity and invasion abilities of Huh7cells were measured by methyl thiazol tetrazolium （MTT） and Transwell assays. The expression levels of proliferating cell nuclear antigen （PCNA） and metalloproteinase- 9 （MMP- 9） were detected by Western blotting.Results miR- 942 was significantly up-regulated in HCC tissues as compared with matched peritumoral tissues （0.92±0.08 vs. 0.42±0.05, t=5.712, P=0.000）. The results of cell proliferation showed that the absorbance （A） value of Huh7 cells （A） in miR- 942 inhibitors group was significantly lower than that of the negative control group and the miR- 942 mimics group after 5 days （0.79±0.02 vs. 1.02±0.04, t=7.513, P=0.001; 0.79±0.02 vs. 1.51±0.12, t=8.621, P=0.001）. The results of Transwell assays showed that the number of metastatic cells in the miR- 942 inhibitors group was significantly decreased compared to the negative control group and the miR- 942 mimics group respectively [（59.13±5.41） cells vs. （97.26±11.13） cells, t=11.712, P=0.000; （59.13±5.41） cells vs. （99.42±12.13） cells, t=12.113, P=0.000]. The expression of PCNA and MMP- 9 protein in miR- 942 inhibitors group was reduced by 80.1%and56.3% respectively （t=5.231, P=0.000; t=6.415, P=0.000）.Conclusion The expression of miR- 942 in HCC tissues was significantly higher than that in adjacent tissues, and dysregulation of miR- 942 can suppress proliferation and invasion of HCC cells, which may be related to down-regulation of PCNA and MMP- 9 expression.
作者
李晓丽
林锐
党晓卫
Li Xiaoli, Lin Rui, Dang Xiaowei(Department of Geriatrics, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, China ;Department of Gastroenterology, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, China ;Department of Hepatobiliary and Pancreatic Surgery, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2018年第3期406-408,共3页
Chinese Journal of Experimental Surgery
基金
河南省科技攻关计划项目（162102310154）
河南省医学科技攻关计划项目（201503033）
关键词
癌
肝细胞
微小RNA-942
增殖
侵袭
Carcinoma, hepatocellular
MicroRNA-942
Proliferation
Invasion
