CRKII overexpression promotes the in vitro proliferation, migration and invasion potential of murine hepatocarcinoma Hca‑P cells

Zhou,Zanmei; Sun,Xiuyan; Guo,Chunmei; Sun,Ming‑Zhong; Liu,Shuqing

doi:10.3892/ol.2019.10194

CRKII overexpression promotes the in vitro proliferation, migration and invasion potential of murine hepatocarcinoma Hca‑P cells

Authors:
- Zanmei Zhou
- Xiuyan Sun
- Chunmei Guo
- Ming‑Zhong Sun
- Shuqing Liu
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Affiliations: Department of Biochemistry, Dalian Medical University, Dalian, Liaoning 116044, P.R. China, Department of Laboratory Medicine, The Affiliated Hospital of Dalian University, Dalian, Liaoning 114012, P.R. China, Department of Biotechnology, Dalian Medical University, Dalian, Liaoning 116044, P.R. China
Published online on: March 27, 2019 https://doi.org/10.3892/ol.2019.10194
Pages: 5169-5174

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Abstract

Lymphatic metastasis is a major mechanism of tumor metastasis. The present study aimed to investigate the association of CRKII, a member of the CRK family, with the in vitro malignant behaviors of a murine hepatocarcinoma Hca‑P cell line, with a lymph node metastatic rate of ~25%. Total mRNA was extracted from Hca‑P cells, and then the murine CRKII gene was amplified by polymerase chain reaction and cloned into the pEASY‑blunt cloning vector. Subsequently, the recombinant pcDNA3.1/V5‑HisB‑CRKII plasmid was constructed and transfected into Hca‑P cells. Western blotting indicated that the CRKII expression level in pcDNA3.1/V5‑HisB‑CRKII‑Hca‑P cells was increased by ~185%, compared with pcDNA3.1/V5‑HisB‑Hca‑P cells. The stable overexpression of CRKII enhanced the in vitro proliferation ability, as measured with a Cell Counting Kit‑8 assay, and the colony forming capacity was measured with a soft agar colony forming assay for Hca‑P cells. The in vitro migration and invasion capacities of Hca‑P cells were increased by ~179 and 156% in Hca‑P cells, respectively, following the stable upregulation of CRKII. Collectively, the recombinant pcDNA3.1/V5‑HisB‑CRKII‑Hca‑P plasmid was constructed successfully. Additionally, the CRKII expression level was positively associated with the in vitro proliferation, migration and invasion malignant properties of Hca‑P cells.

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