Global DNA 5hmC and CK195hmC+ Contents: A Promising Biomarker for Predicting Prognosis in Small Hepatocellular Carcinoma

Background: 5-Hydroxymethylcytosine (5hmC) with dynamic existence possesses multiple regulatory functions. Whereas, 5hmC’s impact on small hepatocellular carcinoma (SHCC) remains unclear. The present work focused on characterizing 5hmC content within SHCC and assessing the possibility of using global genomic 5hmC level as the predicative factor of clinical outcome. Methods: This study applied ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) in measuring 5mC, 5fC and 5hmC contents. In addition, immunohistochemistry (IHC) was adopted to measure CK19 and 5hmC contents. Results: Research showed 5mC, 5hmC, and 5fC contents from global genomics of SHCC reduced extensively compared with healthy samples (p < 0.001). Moreover, SHCC was associated with lymph node metastasis (LNM). Greater 5mC and 5hmC levels were observed in non-metastasis group compared with the metastasis group (p < 0.001). Correlation analysis between the HBV DNA level and 5mC, 5fC and 5hmC levels exhibited that HBV DNA was associated with 5mC, 5hmC, and 5fC content reduction, which was verified in the cytological experiments. Moreover, 5hmC content had a negative correlation with the expression level of CK19 in SHCC. The decrease in 5hmC and CK19 containing 5hmC positive cell (called CK195hmC+) should be ascribed to the bad prognosis among SHCC patients. Conclusions: The contents of 5hmC and CK195hmC+ of genomic DNA might be adopted for predicting SHCC survival as an important biomarker.

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Global DNA 5hmC and CK19 with 5hmC positives cell contents represent a promising biomarker for predicting prognosis in small hepatocellular carcinoma

10.21203/rs.3.rs-21981/v1 ◽

2020 ◽

Author(s):

Jinhua Jiang ◽

Tinghua Yan ◽

Dan Cui ◽

Jun Wang ◽

Fang Guo ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Genomic Dna ◽

High Performance ◽

Clinical Results ◽

Hbv Dna ◽

Small Hepatocellular Carcinoma ◽

Tumor Tissues ◽

Ck19 Expression ◽

Liquid Chromatography Tandem Mass ◽

Regulatory Functions

Abstract Background: 5-Hydroxymethylcytosine (5hmC) exists dynamically and exhibits various regulatory functions. It’s possibly associated with tumor occurrence and malignant transformation. Nevertheless, the part of 5hmC in small hepatocellular carcinoma (SHCC) is still elusive. The study was directed toward characterizing 5hmC content in SHCC and assessing if global genomic 5hmC content was able to serve as a promising factor for predicting clinical results.Methods: The expression contens of 5mC, 5hmC and 5fC were assesed using ultra-high performance liquid chromatography tandem mass-spectrometry(UHPLC-MS/MS). 5hmC contens and CK19 expression were measured by immunohistochemistry(IHC) .Results: The findings in the study displayed that global genomic 5mC, 5hmC, and 5fC contents in SHCC specimens were globally reduced relative to para-tumor tissues (P<0.001), Lymph node metastasis may be found in the small HCC, the non-metastasis group exhibited higher 5mC and 5hmc contents relative to those in metastasis group (P<0.001). HBV DNA was relevant to the decrease in 5mC, 5hmC and 5fC, as evidenced by the measurements in cell lines carrying or not carrying HBV DNA. Moreover, the correlation analysis showed the negative correlation of the content of 5hmC with CK19 expression in SHCC. The decreases of both 5hmC and CK19 with 5hmC positives cell contents in genomic DNA were related to SHCC patients’ unfavorable prognosis. Compared with para-tumor tissues, the 5hmC content in SHCC specimems was dramatically reduced.Conclusions: 5hmC and CK19 with 5hmC positives cell contents in genomic DNA possibly serve as a promising biomarker for predicting prognosis in small hepatocellular carcinoma.

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