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Nogo-C contributes to HCC tumorigenesis via suppressing cell growth and its interactome analysis with comparative
Xing Liu1, Shu-Jian Cui2, Shi-Jun Zhu3
1State Key Laboratory of Genetic Engineering, Fudan University Shanghai, China ; National Engineering Center for Biochip at Shanghai Shanghai, China.
Objects:
Neurite outgrowth inhibitor proteins (Nogos) comprise a family of three major members and are characterized by a conserved RHD domain. Among all the members, Nogo-B was identified to be significantly elevated and to play an important role in liver cirrhosis while Nogo-C was the shortest one and received little attention. The aim of this study is to investigate the relevance and mechanism of Nogo-C involved in Hepatocellular carcinoma (HCC).
Methods:
The expression of Nogo-C in paired HCC specimens was measured with quantitative RT-PCR. The function of Nogo-C over expressing in SMMC-7721 and WRL-68 HCC cell lines were estimated through cell proliferation assay and colony formation assay. A proteome-wide identification of Nogo-C-binding proteins was performed using affinity purification combined with a highly sensitive mass spectrometric technique. The protein interactions were confirmed using co-IP and immunofluorescence confocal assays.
Results:
Compared with the neighboring pathologically normal tissues, the expression of Nogo-C mRNA was extremely down-regulated in HCC specimens and was significantly related to greater tumor size and worse prognosis. Overexpression of Nogo-C in HCC cell lines resulted in an inhibition of cell growth. A total of 73 proteins were detected and considered in association with Nogo-C, among which B-raf and Nogo-B were validated.
Conclusion:
We identify Nogo-C as a tumor suppressor gene in HCC and B-raf as a novel interacting protein. These findings provide new directions for the mechanism research of Nogo family.
Insights
Nogo-C acts as a tumor suppressor in hepatocellular carcinoma (HCC), with its decreased expression linked to worse prognosis. Overexpressing Nogo-C inhibits HCC cell growth, revealing new therapeutic avenues.
Area of Science:
- Molecular Biology
- Oncology
- Biochemistry
Background:
- Neurite outgrowth inhibitor proteins (Nogos) are a family with conserved RHD domains.
- Nogo-B is implicated in liver cirrhosis, while Nogo-C has been less studied.
- Hepatocellular carcinoma (HCC) is a major global health concern.
Purpose of the Study:
- To investigate the role and mechanism of Nogo-C in Hepatocellular Carcinoma (HCC).
- To determine if Nogo-C functions as a tumor suppressor or oncogene in HCC.
- To identify proteins interacting with Nogo-C in HCC.
Main Methods:
- Quantitative RT-PCR to measure Nogo-C mRNA expression in HCC tissues.
- Cell proliferation and colony formation assays to assess Nogo-C function.
- Affinity purification coupled with mass spectrometry to identify Nogo-C binding proteins.
Main Results:
- Nogo-C mRNA expression was significantly downregulated in HCC specimens, correlating with larger tumor size and poorer prognosis.
- Overexpression of Nogo-C inhibited HCC cell growth in vitro.
- B-raf and Nogo-B were identified as Nogo-C interacting proteins.
Conclusions:
- Nogo-C acts as a tumor suppressor gene in HCC.
- B-raf is a novel interacting protein with Nogo-C.
- These findings offer new insights into the Nogo family's mechanism in cancer.
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