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Overexpressed miR-301a promotes cell proliferation and invasion by targeting RUNX3 in gastric cancer

  • Original Article—Alimentary Tract
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Abstract

Background

MicroRNAs can promote or suppress the evolution of malignant behaviors by regulating multiple targets. We aimed to determine the expression of miR-301a recently screened in gastric cancer, to investigate the biological effects of miR-301a and to identify the specific miR-301a target gene.

Methods

Quantitative real-time RT-PCR was used to test miR-301a expression. Functional effects were explored by a water-soluble tetrazolium salt assay, a colony formation assay in soft agar, a migration assay, an invasion assay and cytometry used to determine apoptosis and cell cycle. Nude mice were inoculated subcutaneously by retrovirus-mediated stably expressed SGC-7901 cells. The target gene was determined by bioinformatic algorithms, dual luciferase reporter assay and Western blot.

Results

Firstly, we found that miR-301a was significantly upregulated both in cells and tissues of gastric cancer. The expression level of miR-301a was inversely correlated with tumor differentiation of gastric cancer tissues. Secondly, miR-301a promoted cell growth, soft agar clonogenicity, migration, invasion, and decreased cell apoptosis induced by cisplatin in vitro, while blockage of miR-301a reduced the percentage of G2/M phase cells via flow cytometry in gastric cancer cells. Ectopic expression of miR-301a enhanced the subcutaneous tumorigenesis in vivo. Finally, miR-301a directly downregulated RUNX3 expression post-transcriptionally in gastric cancer.

Conclusion

Our results demonstrate that miR-301a plays important roles in the development of gastric cancer.

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Acknowledgments

This study was supported by grants from the National Natural Science Foundation of China (No. 30872476, No. 30900670, No. 81101585, No. 91229106 and No. 81172324), and Science and Technology Commission of Shanghai Municipality (No. 12XD1403700, and No. 12PJ1406300).

Conflict of interest

None declared.

Author information

Author notes

  1. Ming Wang

    Present address: Department of Gastrointestinal Surgery, Heze Municipal Hospital, 2888 # Caozhou Road, Heze, 274000, Shandong, People’s Republic of China

Authors and Affiliations

  1. Shanghai Key Laboratory of Gastric Neoplasms, Department of Surgery, Shanghai Institute of Digestive Surgery, Ruijin Hospital, School of Medicine, Shanghai Jiao Tong University, 197# Ruijin Er Road, Shanghai, 200025, People’s Republic of China

    Ming Wang, Chenglong Li, Beiqin Yu, Liping Su, Jianfang Li, Yingyan Yu, Qinlong Gu, Zhenggang Zhu & Bingya Liu

  2. Department of Pathology, Stony Brook University Medical Center, Stony Brook, NY, 11794-8691, USA

    Jingfang Ju

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  1. Ming Wang
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Correspondence to Bingya Liu.

Electronic supplementary material

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S1 Level of miR-301a expression was upregulated in 88 % (45/51) of gastric cancer tissues. (TIFF 525 kb)

535_2012_733_MOESM2_ESM.tif

S2 The miR-301a expression in SGC-7901 cells was effectively altered by transient transfection of miR-301a (miR-301a mimics) as detected by RT-PCR. U6 snRNA was used for normalization. (TIFF 429 kb)

535_2012_733_MOESM3_ESM.tif

S3 The miR-301a expression in SGC-7901 cells was effectively altered by transient transfection of anti-miR-301a (miR-301a inhibitors) as detected by RT-PCR. (TIFF 434 kb)

S4 Expression of miR-310a was detected in different cell cycles using RT-PCR. (TIFF 138 kb)

S5 miR-301a expression level in SGC-7901/retro-miR-301a or SGC-7901/vector was analyzed by qRT-PCR. (TIFF 386 kb)

535_2012_733_MOESM6_ESM.tif

S6 RUNX3 mRNA expression in SGC-7901 cells was analyzed by qRT-PCR 48 h post-transfection with miR-301a precursor and control precursor (100 nM). The results was shown as fold change relative to the control precursor-transfected SGC-7901 cells. (TIFF 541 kb)

S7 RUNX3 expression level in SGC-7901 cells was analyzed by qRT-PCR. (TIFF 223 kb)

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Wang, M., Li, C., Yu, B. et al. Overexpressed miR-301a promotes cell proliferation and invasion by targeting RUNX3 in gastric cancer. J Gastroenterol 48, 1023–1033 (2013). https://doi.org/10.1007/s00535-012-0733-6

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  • DOI: https://doi.org/10.1007/s00535-012-0733-6

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