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[微小RNA-30a-5p对人肝癌细胞SMCC-7721增殖、凋亡、侵袭及迁移的影响]

[Effect of miR-30a-5p on the proliferation, apoptosis, invasion and migration of SMCC-7721 human hepatocellular carcinoma cells].

作者信息

Dai Hang, Kang Bing, Zuo Deyu, Zuo Guoqing

机构信息

Department of Digestive System, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.

出版信息

Zhonghua Gan Zang Bing Za Zhi. 2014 Dec;22(12):915-20. doi: 10.3760/cma.j.issn.1007-3418.2014.12.008.

Abstract

OBJECTIVE

To explore the effect of microRNA-30a-5p (miRNA-30a-5p) on the biological behavior of human hepatoma cells.

METHODS

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.

RESULTS

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).

CONCLUSION

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.

摘要

目的

探讨微小RNA-30a-5p(miRNA-30a-5p)对人肝癌细胞生物学行为的影响。

方法

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。

结果

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)。

结论

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。

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