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Hypermethylation of ACP1, BMP4, and TSPYL5 in Hepatocellular Carcinoma and Their Potential Clinical Significance

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Abstract

Background and Aim

Aberrant methylation of specific genes is frequent event in hepatocellular carcinoma (HCC). Our present study aims to explore the methylation levels of acid phosphatase locus 1 (ACP1), bone morphogenetic protein 4 (BMP4), and testis-specific protein, Y-encoded-like 5 (TSPYL5) and their potential clinical applications in HCC.

Methods

The methylation levels of ACP1, BMP4 and TSPYL5 were analyzed in 188 HCC tissues, 163 matched adjacent non-tumor tissues, and 29 normal liver tissues using a method of methylation-sensitive restriction enzyme-based quantitative PCR, and their associations with clinicopathological features and prognosis were evaluated.

Results

Compared with adjacent non-tumor tissues and normal liver tissues, the methylation levels of ACP1, BMP4, and TSPYL5 were significantly increased in HCC tissues (All p < 0.0001). The methylation of each individual gene could distinguish HCC tissues well from adjacent non-tumor tissues with the area under the receiver operating characteristic curves (AUC) of 0.753, 0.785 and 0.917, respectively. Furthermore, a higher methylation of BMP4 was statistically associated with worse disease-free survival (p = 0.006) and might be an independent unfavorable factor for disease-free survival by univariate and multivariate analysis (p = 0.011, HR 3.431, 95 % CI 1.333–8.833).

Conclusions

Our findings suggest that hypermethylation of ACP1, BMP4, and TSPYL5 are common events in HCC and could be used as potentially detectable biomarkers in HCC tissues. Moreover, BMP4 could be potentially served as a methylated biomarker to predict recurrence and metastasis after hepatectomy for HCC patients. However, their potential clinical application value need to be further clarified.

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Acknowledgments

This study was supported by the National Basic Research Program of China (973 Program, 2012CB720605), the Science and Technology Research Plan of Wuhan City (2015060101010057), and Guangdong province production Scientific Research Project (2013B090500101).

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    Authors and Affiliations

    1. Center for Gene Diagnosis, Zhongnan Hospital of Wuhan University, Donghu Rd 169, Wuchang District, Wuhan, 430071, China

      Xueping Qiu, Yifang Huang, Xuebin Wang & Fang Zheng

    2. The Third Affiliated Hospital of Sun Yat-sen University, Guanzhou, Guandong, China

      Bo Hu

    3. Department of Pathology, Hubei Cancer Hospital, Wuhan, Hubei, China

      Yunte Deng

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    1. Xueping Qiu
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    Correspondence to Fang Zheng.

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    Xueping Qiu and Bo Hu have contributed equally to this work.

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    Supplemental Data, Fig. S1

    Representative DNA methylation status of three genes in HCC and paired adjacent non-tumor tissues. (a, b, c, d, e, f) Methylation status of ACP1, BMP4 and TSPYL5 in HCC and paired adjacent non-tumor tissues by BGS. Sequencing for randomly ten clones of PCR products amplified from each sample was shown by black and white circles, which represent methylated and unmethylated CG dinucleotides, respectively. Red circles indicate the CpG sites within recognition sites of MSRE. Each row represents the sequencing result of one clone, and the number represents the position of CpG sites in target regions (number 1 represent the first CpG site in the target region). MSRE–qPCR results for the same sample of BGS were displayed as amplification curve (g, i, k) and melt curve (h, j, l). The blue and purple line indicated digested sample for HCC and paired adjacent non-tumor tissues, respectively. The red and green line indicated undigested sample for HCC and paired adjacent non-tumor tissues, respectively. HCC: hepatocellular carcinoma tissues; NT: adjacent non-tumor tissues (TIFF 1238 kb)

    Supplemental Data, Table S1

    Primers used in the study. (DOC 35 kb)

    Supplemental Data, Table S2

    Associations between DNA methylation levels and clinicopathologic parameters. (DOC 91 kb)

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    Qiu, X., Hu, B., Huang, Y. et al. Hypermethylation of ACP1, BMP4, and TSPYL5 in Hepatocellular Carcinoma and Their Potential Clinical Significance. Dig Dis Sci 61, 149–157 (2016). https://doi.org/10.1007/s10620-015-3878-3

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    • DOI: https://doi.org/10.1007/s10620-015-3878-3

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