Abstract
Purpose
Circular RNAs (circRNA) represent a novel class of widespread and diverse endogenous RNAs that regulate gene expression in mammals. microRNA-7 (miR-7) is a well-demonstrated suppressor of hepatocellular carcinoma (HCC). Recent studies have showed that one such circRNA, ciRS-7 (also termed as Cdr1as) was the inhibitor and sponge of miR-7 in the embryonic zebrafish midbrain and islet cells. However, the relationships among ciRS-7, miR-7 and clinical features of HCC remain to be clarified.
Methods
Expression levels of ciRS-7, miR-7 and three miR-7-targeted mRNAs in 108 pairs of HCC and their matched non-tumor tissues were examined by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The protein production of these three miR-7-targeted mRNAs was further verified by Western blot. The relationship between ciRS-7 level and clinicopathological features as well as the recurrence of HCC patients was analyzed. The univariate and multivariate logistic regression analyses were used to detect the risk factors of hepatic microvascular invasion (MVI). The correlation among ciRS-7, miR-7 and miR-7-targeted mRNAs was evaluated using Spearman’s correlation test.
Results
There was no significant difference of ciRS-7 expression levels between the HCC tissues and the matched non-tumor tissues (0.67 ± 1.49 vs. 0.44 ± 0.45, p = 0.13), and the ciRS-7 levels in more than half of HCC tissues (65 out of 108, 60.2 %) were down-regulated when compared with their matched non-tumor tissues. However, the expression of ciRS-7 was significantly correlated with the following three clinicopathological characteristics of HCC patients: age <40 years (p = 0.02), serum AFP ≥400 ng/µl (p < 0.01) and hepatic MVI (p = 0.03). Meanwhile, up-regulated ciRS-7 expression was not only an independent risk factor of hepatic MVI but also had a capable predictive ability for MVI (AUC = 0.68, p = 0.001) at the cut-off value of 0.135. Furthermore, the expression of ciRS-7 in HCC tissues with concurrent MVI was inversely correlated with that of miR-7 (r = −0.39, p = 0.007) and positively related with that of two miR-7-targeted genes [PIK3CD (r = 0.55, p < 0.001) and p70S6K (r = 0.34, p = 0.021)]. In addition, the median recurrent time of patients from higher ciRS-7 level group was shorter than that of lower ciRS-7 group (18 vs. 25 months), but no significant difference was observed (p = 0.38).
Conclusions
The expression levels of ciRS-7 were comparable between HCC and matched non-tumor tissues. However, the highly ciRS-7 expression in HCC tissues was significantly correlated with hepatic MVI, AFP level and younger age and thus partly related with the deterioration of HCC. Especially, ciRS-7 was one of the independent factors of hepatic MVI. These data suggested that ciRS-7 may be a promising biomarker of hepatic MVI and a novel therapy target for restraining MVI.
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Abbreviations
- HCC:
-
Hepatocellular carcinoma
- AFP:
-
Alpha fetal protein
- MVI:
-
Microvascular invasion
- GVI :
-
Gross vascular invasion
- ciRS-7:
-
Circular RNA sponge for microRNA-7
- CDR1as:
-
Cerebellar degeneration-related protein 1 antisense RNA
- miR-7:
-
microRNA-7
- qRT-PCR:
-
Quantitative reverse transcription polymerase chain reaction
- VDAC1:
-
Voltage dependent anion channel 1
- ROC:
-
The receiver operating characteristic
- AUC:
-
Area under the curve
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Acknowledgments
This study was funded by the National Natural Science Foundation of China (No. 71673193), and Key Technology Research and Development Program of the Sichuan Province (2015SZ0131).
Authors’ contributions
MX, L.X, and X.Z conceived and designed the study; L.X, P.Y, C.L performed all experiments; Z.M responsible for the analysis and interpretation of data. L.X and M.Z wrote the paper.
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The authors declared that they have no conflicts from commercial interest.
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All procedures performed in this study involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards. Permission was gained from the Biomedical Ethics Committee of West China Hospital for this retrospective analysis.
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Written informed consent for the use of tissue samples for scientific purposes was obtained from all individual participants included in the study before the operation.
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Liangliang Xu and Ming Zhang have contributed equally to this study.
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Xu, L., Zhang, M., Zheng, X. et al. The circular RNA ciRS-7 (Cdr1as) acts as a risk factor of hepatic microvascular invasion in hepatocellular carcinoma. J Cancer Res Clin Oncol 143, 17–27 (2017). https://doi.org/10.1007/s00432-016-2256-7
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DOI: https://doi.org/10.1007/s00432-016-2256-7