shRNA-mediated silencing of the RFC3 gene suppresses hepatocellular carcinoma cell proliferation.

Zhicheng Yao, Kunpeng Hu, He Huang, Shilei Xu, Qingliang Wang, Peng Zhang, Peisheng Yang, Bo Liu

International journal of molecular medicine 2015 Nov

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Hepatocellular carcinoma (HCC) is one of the most common and lethal malignancies worldwide. Replication factor C (RFC) plays an important role in DNA replication and checkpoint control during the cell cycle. RFC is comprised of one large subunit [replication factor C, subunit 1 (RFC1)] and four small subunits [replication factor C, subunits 2-5 (RFC2‑5)]. The role of RFC3 in the development of HCC is, as of yet, not fully understood. In the present study, western blot analysis and reverse-transcription-quantitative PCR were used to measure the expression levels of replication factor C, subunit 3 (RFC3) in HCC tissues and HCC cells. Lentivirus-mediated RFC3-specific short hairpin RNA (shRNA) was used to knock down the expression of RFC3 in HCC cells in order to examine the effects of RFC3 on HCC cell proliferation and growth. Furthermore, the expression levels of cell cycle‑related proteins were also measured in the HCC cells in which RFC3 was knocked down. Our results revealed that the expression level of RFC3 was markedly upregulated in the HCC tissues and cells. In addition, MTS and cell growth assays were used to determine the viability and proliferation of the HCC cells in which RFC3 was knocked down, and the results revealed that both cell viability and proliferation were effectively suppressed. The downregulation of RFC3 expression led to HCC cell cycle arrest in the S phase, partly by regulating the epression of cell cycle-related proteins, such as p21, p53, p57 and cyclin A. The results of the present study suggest that RFC3 plays an important role in the development of HCC, and may thus be a potential biological target in the treatment of HCC.

Citation

Zhicheng Yao, Kunpeng Hu, He Huang, Shilei Xu, Qingliang Wang, Peng Zhang, Peisheng Yang, Bo Liu. shRNA-mediated silencing of the RFC3 gene suppresses hepatocellular carcinoma cell proliferation. International journal of molecular medicine. 2015 Nov;36(5):1393-9