Dai Hang, Kang Bing, Zuo Deyu, Zuo Guoqing
Department of Digestive System, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.
Zhonghua Gan Zang Bing Za Zhi. 2014 Dec;22(12):915-20. doi: 10.3760/cma.j.issn.1007-3418.2014.12.008.
To explore the effect of microRNA-30a-5p (miRNA-30a-5p) on the biological behavior of human hepatoma cells.
The liver cancer cell line SMCC-7721 cells and the normal liver cell line L02 cells (control) were transiently transfected with miRNA-30a-5p mimics and an miRNA-30a-5p inhibitor by Lipofectamine 2000 (Life Technologies). miR-30a-5p mRNA expression was detected by quantitative real-time (q)PCR. Cell proliferation was evaluated using the Cell Counting Kit-8 (CCK-8) assay and apoptosis was assessed by flow cytometry.Invasion and migration were measured by transwell chamber assays. The SMCC-7721 cells was injected subcutaneously into nude mice to establish a tumor animal model.
The SMCC-7721 cells transfected with miRNA-30a-5p mimics showed significantly higher miRNA-30a-5p mRNA expression than the non-transfected SMCC-7721 cells and the transfected control L02 cells (P<0.01). The miRNA-30a-5p mRNA expression was significantly lower in the SMCC-7721 cells transfected with the miRNA-30a-5p inhibitor than the non-transfected SMCC-7721 cells the control L02 cells (P<0.01). The overexpression of miRNA-30a-5p inhibited the viability, colony formation rate, and invasion and migration abilities, as shown in the cells transfected with the miRNA-30a-5p mimics (P<0.05); in addition, the miRNA-30a-5p promoted proliferation of cells (P<0.05), as shown by more S phase cells detected by flow cytometry. SMCC-7122 cells transfected with miRNA-30a-5p mimics produced tumors with significantly higher average weight than tumors produced by SMCC-7122 cells that were untransfected or transfected with empty vector (both P<0.01).
Overexpression ofmiR-30a-5p had an inhibitory effect on cell proliferation, induced apoptosis, increased the number of cells in S phase, and markedly inhibited invasion and migration of SMCC-7721 HCC cells in vitro and in vivo.
探讨微小RNA-30a-5p（miRNA-30a-5p）对人肝癌细胞生物学行为的影响。
采用脂质体2000（赛默飞世尔科技公司）将miRNA-30a-5p模拟物和miRNA-30a-5p抑制剂分别瞬时转染肝癌细胞系SMCC-7721细胞和正常肝细胞系L02细胞（对照）。通过定量实时荧光定量PCR检测miR-30a-5p mRNA表达。使用细胞计数试剂盒-8（CCK-8）检测法评估细胞增殖，采用流式细胞术评估细胞凋亡。通过Transwell小室检测法测定细胞侵袭和迁移能力。将SMCC-7721细胞皮下注射到裸鼠体内建立肿瘤动物模型。
转染miRNA-30a-5p模拟物的SMCC-7721细胞的miRNA-30a-5p mRNA表达明显高于未转染的SMCC-7721细胞和转染对照L02细胞（P<0.01）。转染miRNA-30a-5p抑制剂的SMCC-7721细胞的miRNA-30a-5p mRNA表达明显低于未转染的SMCC-7721细胞和对照L02细胞（P<0.01）。如转染miRNA-30a-5p模拟物的细胞所示，miRNA-30a-5p的过表达抑制了细胞活力、集落形成率以及侵袭和迁移能力（P<0.05）；此外，如流式细胞术检测到更多处于S期的细胞所示，miRNA-30a-5p促进了细胞增殖（P<0.05）。转染miRNA-30a-5p模拟物的SMCC-7122细胞产生的肿瘤平均重量明显高于未转染或转染空载体的SMCC-7122细胞产生的肿瘤（均P<0.01）。
miR-30a-5p的过表达对细胞增殖具有抑制作用，诱导细胞凋亡，增加S期细胞数量，并在体外和体内显著抑制SMCC-7721肝癌细胞的侵袭和迁移。
