Homer2 and Homer3 Act as Novel Biomarkers in Diagnosis of hepatitis B virus-induced Hepatocellular Carcinoma

Abstract

Background: Hepatocellular carcinoma (HCC) is one of the most common causes of cancer-related mortality worldwide. Early detection of HCC can significantly improve patients' outcomes. An increasing number of studies have validated that Homer is dysregulated in cancers and may serve as diagnostic markers. In the present study, we investigated the expression profile and diagnostic significance of Homer2 and Homer3 in hepatitis B virus-induced HCC (HBV-HCC). Methods: Quantitative real-time PCR (QRT-PCR), western blot analysis and immunohistochemistry analysis. Results: Homer2 and Homer3 were downregulated in HCC. The expression of Homer2 was associated with tumor differentiation grade (P= 0.012) and total protein (TP) level (P= 0.032). Homer3 was related to tumor size (P= 0.010), tumor nodes (P= 0.026) and γ-glutamyl transferase (GGT) level (P= 0.001). The receiver operating characteristic curve analyses indicated that the combination of Homer2, Homer3 and AFP possessed a high accuracy (AUC=0.900) to diagnose HCC cases from healthy controls. Conclusion: Our data indicated that Homer2 and Homer3 were downregulated in HCC and might be potential diagnostic marker for HCC.

Keywords: Diagnosis.; HCC; Homer2; Homer3; Tumorigenesis.

Figure 1

Evaluation of Homer2 and Homer3 expression in HCC cell lines. Expression of Homer2 in HCC cell lines were lower than that in L-02 cell (A, C, E). Expression of Homer3 in HCC cell lines were lower than that in L-02 cells (B, D, F). * P< 0.05.

Figure 2

Homer2 and Homer3 expression in HCC tissues. The relative Homer2 and Homer3 expression were determined by RT-qPCR. Δ Ct values were calculated by subtracting the GAPDH Ct value from the Homer2 or Homer3 Ct value. Smaller ΔΔ Ct value indicates higher expression. ΔΔ Ct = (Ct _{Homer2/Homer3}-Ct _GAPDH) of HCC - (Ct _{Homer2/Homer3}- Ct _GAPDH) of NT]. (NT: paired noncancerous tissues of HCC). (A) Levels of Homer2 in tumor tissues are significantly lower than that in non-tumor tissues. (B) Levels of Homer3 in tumor tissues are significantly lower than that in non-tumor tissues. (C) Homer2 was reduced by at least twofold in 53.2% (41/77) of the HCC tissues. (D) Homer3 was downregulated in 62.3% (48/77) of HCC tissues. All data were analyzed using Student's t test. Data were presented as mean ± SD, * P < 0.05, ** P< 0.01.

Figure 3

Homer2 and Homer3 expression in peripheral blood among subgroups. (A) Homer2 expression in HCC was lower than that in cirrhosis, hepatitis B and the controls. No differences were observed among other subgroups. (B) Homer3 expression in HCC was lower than that in cirrhosis, hepatitis B and the controls and Homer3 expression in cirrhosis and hepatitis B were lower than that in controls. No differences were observed between hepatitis B and cirrhosis. Data were analyzed using oneway ANOVA. * P < 0.05, ** P < 0.01.

Figure 4

The expression levels of Homer2 and Homer3 in 14 paired preoperative and postoperative peripheral blood samples. (A) Levels of Homer2 was increased 2 weeks after surgical (P=0.024); (B) Levels of Homer3 was increased 2 weeks after surgical (P=0.022).

Figure 5

Receiver operating characteristic (ROC) curves. (A) Homer2 and Homer3 for HCC vs Controls; (B) Homer2 and Homer3 for HCC vs HBV; (C) Homer2 and Homer3 for HCC vs patients with cirrhosis; (D) the combination of Homer2 and Homer3 for HCC vs Controls; (E) the combination of Homer2, Homer3 and AFP for HCC vs Controls.