Screening the effect targets of CX43 in hepatocellular carcinoma tissues with whole transcriptome sequencing

doi:10.3877/cma.j.issn.2095-3232.2019.02.018

Abstract

Abstract:

Objective

To screen the downstream differentially-expressed genes of gap junction protein CX43 by using the whole transcriptome sequencing combined with bioinformatics and to analyze its function.

Methods

Human liver cancer SMMC-7721 cells were transfected with recombinant pCX43 plasmid with TransIntro^TM EL to construct the CX43 over-expression group, and the cells were transfected with empty plasmid in control group. The expression of CX43 mRNA and protein level were quantitatively measured by RT-PCR and Western blot, and were compared between two groups by t test. The downstream differentially-expressed genes of CX43 were screened by whole transcriptome Clariomtm D Assays. The gene function, signaling pathway and transactional function of protein in differentially-expressed genes were analyzed with DAVID and STRING databases to determine the central node proteins. The predictive value of target protein in survival and prognosis was analyzed with Kaplan-Meier curve and Cox's regression analysis.

Results

The expression levels of CX43 mRNA and protein in overexpression group were 363±64 and 1.36±0.02, significantly higher than 1 and 0.81±0.01 in control group (t=5.67,19.12; P<0.05). The downstream differentially-expressed genes of CX43 were screened, of which 394 genes were up-regulated and 534 were down-regulated. 9 central node proteins were analyzed by STRING Database. Kaplan-Meier curve and Cox's regression analysis demonstrated that the overall survival of patients with high expression of ENO1, RALA and SRC was poor (HR=2.29, 1.72, 1.75; 95%CI: 1.61-3.27, 1.20-2.46, 1.22-2.53; P<0.05).

Conclusions

ENO1, RALA and SRC are the downstream effect targets of CX43. Three target proteins are the independent factors affecting the survival and prognosis. Patients with high expression of target proteins have poor clinical prognosis.

Key words: Carcinoma, hepatocellular, Connexin 43, Microchip analytical procedures

Zhen Tan, Weidong Guo, Lu Zhang, Zehua Wu, Shixiong Zhan, Shiguo Liu, Zusen Wang. Screening the effect targets of CX43 in hepatocellular carcinoma tissues with whole transcriptome sequencing[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2019, 08(02): 164-168.

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References 20

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