Expression of RNFT2 in hepatocellular carcinoma patients and effect on cell proliferation, invasion and prognosis

doi:10.3877/cma.j.issn.2095-3232.2025.04.018

Abstract

Abstract:

Objective

To investigate the expression level of ring finger protein, transmembrane 2 (RNFT2) in hepatocellular carcinoma (HCC) and its effect on cell proliferation, invasion and prognosis.

Methods

The expression levels of RNFT2 in HCC, normal liver and HCC tissues with different tumor stages and pathological grades were obtained by using UALCAN. R2 genomics analysis and visualization platform were used to analyze the difference of RNFT2 expression level between HCC and normal liver tissues. The correlation between the expression of RNFT2 and survival of HCC patients was analyzed by gene expression profile (GepLiver) in liver cells and tissues. Clinical samples were collected from 24 pairs of HCC and adjacent tissues of HCC patients undergoing surgical resection at the First Affiliated Hospital of Xi’an Jiaotong University from January to December 2021. Among them, 19 patients were male and 5 female, aged from 42 to 71 years, with a median age of 52 years. The expression levels of RNFT2 protein in the HCC and adjacent tissues, and normal liver cells and HCC cells were detected by Western blot. The expression levels of RNFT2 in HCCLM3 and Huh7 cells were down-regulated using small interfering RNA (siRNA). The effect of RNFT2 knockdown on the proliferation, migration and invasion of HCC cells was analyzed by EdU-based assay and Transwell chamber test. One-way ANOVA, LSD-t test and t-test were used to analyze the differences among different groups. Survival analysis was performed by Kaplan-Meier method and Log-rank test.

Results

The transcriptome sequencing data of HCC and normal liver tissues in TCGA and GEO databases were analyzed by UALCAN and R2 platforms. The results showed that the expression level of RNFT2 mRNA in HCC tissues was significantly higher than that in normal liver tissues (t=19.343, 7.133, 6.934; all P<0.001). Western blot of 24 pairs of HCC and matched adjacent tissues showed that the relative expression of RNFT2 protein in HCC was 2.1 (1.1, 3.3), significantly higher than 1.1 in adjacent tissues (0.5, 1.7) (Z=-2.536; P<0.05). Analysis of TCGA database by UALCAN platform found that RNFT2 mRNA was associated with tumor staging and pathological grading (F=5.082,8.008; both P<0.05). The expression of RNFT2 mRNA in stage Ⅱ and Ⅲ HCC tissues was significantly higher than that in stage Ⅰ HCC tissues (LSD-t=0.161, 0.162; both P<0.05), and the expression of RNFT2 mRNA in high pathological grade (grade Ⅲ and grade Ⅳ) was significantly higher than that in low pathological grade (grade Ⅰ and grade Ⅱ) (LSD-t=0.196, 0.142, 0.384, 0.360; all P<0.05). GepLiver platform analysis of the survival data of HCC patients in TCGA database showed that high expression of RNFT2 mRNA prompted that the overall survival, disease-specific survival, disease-free survival and progression-free survival of HCC patients were significantly shortened (HR=1.86, 1.73, 1.60, 1.82; all P<0.05). The expression of RNFT2 in HCC cell line was significantly higher than that in normal liver cells (P<0.05). RNFT2 knockdown significantly weakened the proliferation, migration and invasion of HCCLM3 and Huh7 cells (all P<0.05).

Conclusions

The expression level of RNFT2 is up-regulated in HCC tissues, which is associated with poor prognosis of HCC patients. RNFT2 knockdown significantly inhibits the proliferation, migration and invasion of HCC cells.

Key words: Carcinoma, hepatocellular, Ring finger protein, Prognosis, Proliferation, Invasion

Yong Guo, Siying Jia, Shijie Gao, Kangsheng Tu. Expression of RNFT2 in hepatocellular carcinoma patients and effect on cell proliferation, invasion and prognosis[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2025, 14(04): 619-627.

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References 30

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