Construction of a recombinant eukaryotic expression vector containing PHD3 gene and its expression in HepG2 cells

Abstract

Prolyl hydroxylase domain 3 (PHD3) is a hypoxia inducible factor-α (HIFα) regulator; it degrades HIFα in the presence of oxygen. Recently, there have been an increasing number of studies about the role of PHD3 in proliferation and apoptosis of cancer cells. However, most of the evidence for the role of PHD3 is observational, and little is known of the molecular mechanism. In our current study, we constructed a recombinant eukaryotic expression vector containing the PHD3 gene and detected its biological activity in human hepatoma cell line (HepG2 cells). We successfully constructed a recombinant pcDNA 3.1(+)-PHD3 plasmid; the results showed that PHD3 overexpression could inhibit the proliferation of HepG2 cells and induce apoptosis by activating caspase-3 activity. Our study has provided preliminary materials and data for further investigation of the effect of PHD3 on HepG2 cells.

Association of PHD3 and HIF2α gene expression with clinicopathological characteristics in human hepatocellular carcinoma

“…The present study aimed to explore the association between PHD3 and HIF2α expression in HCC (under hypoxic conditions). Several studies have demonstrated that PHD3 serves a novel role in the progression and prognosis of cancer (15-24); PHD3 is also able to induce apoptosis and inhibit proliferation in cancer cells (22)(23)(24)(25)(26)(27)(28)(29). To the best of our knowledge, the present study is the first to report that PHD3 overexpression induces apoptosis and inhibits growth and proliferation in HCC cells.…”

“…Given the increasing number of observational studies on PHD3, considerable attention has been focused on the mechanism of PHD3 (17,18,(24)(25)(26)(27)(28). Su et al (18) revealed that PHD3-induced apoptosis is dependent on nerve growth factor by activating caspase-3 in pancreatic cancer.…”

“…Recent studies have demonstrated that PHD3 suppresses tumor growth by regulating the epidermal growth factor receptor activity (26,27). In our previous study, a vector containing the PHD3 gene was transfected into HepG2 cells and PHD3 overexpression was revealed to inhibit cell proliferation and induce apoptosis by activating caspase-3 (28). Furthermore, stably PHD3-overexpressing HepG2 cells were injected into nude mice and the average tumor size of the PHD3 overexpression group was identified to be larger compared with the control group (25).…”

“…Additionally, PHD3 may be associated with another pathway. Our previous study revealed that PHD3 overexpression could not regulate HIF2α expression in HpG2 cells (28). Furthermore, Tanaka et al (17) demonstrated that PHD3 is able to be regulated by the PI3K/Akt/mTOR pathway independently of HIF proteins in renal cell carcinoma.…”

Prolyl hydroxylase domain protein 3 and asparaginyl hydroxylase factor inhibiting HIF-1 levels are predictive of tumoral behavior and prognosis in hepatocellular carcinoma

“…Another two studies provided further information of human hepatoma cell line. By using PHD3 steady expression plasmid transfected HepG2 and constructing HepG2-bearing subcutaneous tumor in nude mice, researchers found out that PHD3 gene may inhibit proliferation and induce apoptosis by activating caspase-3 activity [ 63 , 64 ]. Taken together, the tumor suppressive effect of FIH and PHD3 should exert through inhibiting proliferation or blocking neovascularization.…”