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微小RNA-320a通过靶向人类肝细胞癌中的c-Myc抑制肿瘤增殖和侵袭。

miRNA-320a inhibits tumor proliferation and invasion by targeting c-Myc in human hepatocellular carcinoma.

作者信息

Xie Fei, Yuan Yuncang, Xie Luyang, Ran Pengzhan, Xiang Xudong, Huang Qionglin, Qi Guoxiang, Guo Xiaopeng, Xiao Chunjie, Zheng Shangyong

机构信息

School of Medicine, Yunnan University, Kunming, Yunnan.

Department of Stomatology, Shanghai Tenth People's Hospital, Shanghai.

出版信息

Onco Targets Ther. 2017 Feb 15;10:885-894. doi: 10.2147/OTT.S122992. eCollection 2017.

Abstract

BACKGROUND

Downregulated expression levels of microRNA-320a (miR-320a) were found in primary breast cancers and colorectal cancer. Previous findings indicated that miRNA-320a may involve in the cancer development. In this study, we explored the roles of miR-320a by targeting c-Myc in the tumor growth of hepatocellular carcinoma (HCC).

METHODS

Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was performed to detect the expression of miR-320a in 50 HCC tissues and four HCC cells. Luciferase reporter assay was conducted to confirm the direct downstream target of miR-320a in HEK-293 cells. The effect of miR-320a on endogenous c-Myc expression was investigated by transfecting miR-320a mimics into HepG2 and QGY-7703 cell lines. The c-Myc and miR-320a expressions were analyzed by immunohistochemistry (IHC) and qRT-PCR in the same HCC tissues. Furthermore, the biological functional correlation of miR-320a with c-Myc was determined by studying the effect of miR-320a mimics or c-Myc small interfering RNA (siRNA) on HCC cell proliferation and invasion.

RESULTS

The expression of miR-320a was downregulated in 50 HCC tissues and 4 HCC cells. Luciferase assay revealed that c-Myc is a direct target of miR-320a. IHC and Western blot analysis showed that the c-Myc expression was inhibited by miR-320a in HCC tissues and cell lines. Upregulation of miR-320a suppressed the HCC cell proliferation and invasion capacity induced by inhibiting c-Myc, and the results were consistent with the effects of c-Myc siRNA on tumor suppression. These results revealed that miRNA-320a inhibits tumor proliferation and invasion by targeting c-Myc in HCC cells.

CONCLUSION

Our results showed that miR-320a functions as a tumor suppressor in HCC. By targeting c-Myc directly, miR-320a inhibits the HCC cell growth. Our studies provide evidence of miR-320a as a potentially target for HCC treatment.

摘要

背景

在原发性乳腺癌和结直肠癌中发现微小RNA-320a(miR-320a)表达水平下调。先前的研究结果表明,miRNA-320a可能参与癌症发展。在本研究中,我们通过靶向c-Myc探讨了miR-320a在肝细胞癌(HCC)肿瘤生长中的作用。

方法

采用定量逆转录聚合酶链反应(qRT-PCR)检测50例HCC组织和4种HCC细胞中miR-320a的表达。进行荧光素酶报告基因检测以证实miR-320a在HEK-293细胞中的直接下游靶点。通过将miR-320a模拟物转染到HepG2和QGY-7703细胞系中,研究miR-320a对内源性c-Myc表达的影响。在同一HCC组织中,通过免疫组织化学(IHC)和qRT-PCR分析c-Myc和miR-320a的表达。此外,通过研究miR-320a模拟物或c-Myc小干扰RNA(siRNA)对HCC细胞增殖和侵袭的影响,确定miR-320a与c-Myc的生物学功能相关性。

结果

50例HCC组织和4种HCC细胞中miR-320a表达下调。荧光素酶检测显示c-Myc是miR-320a的直接靶点。免疫组织化学和蛋白质印迹分析表明,miR-320a在HCC组织和细胞系中抑制c-Myc表达。miR-320a的上调通过抑制c-Myc抑制HCC细胞增殖和侵袭能力,结果与c-Myc siRNA的肿瘤抑制作用一致。这些结果表明,miRNA-320a通过靶向HCC细胞中的c-Myc抑制肿瘤增殖和侵袭。

结论

我们的结果表明,miR-320a在HCC中起肿瘤抑制作用。通过直接靶向c-Myc,miR-320a抑制HCC细胞生长。我们的研究为miR-320a作为HCC治疗的潜在靶点提供了证据。

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