Abstract
Hemocyanin (HMC) has been shown to participate in multiple roles of immune defence. In this study, we investigated the antiproliferative effect and underpinning mechanism of HMC from Litopenaeus vannamei in vitro. Sulforhodamine B (SRB) assay indicated that HMC could dramatically inhibit the growth of HeLa cells, but not 293T cells under the same conditions. Moreover, typical morphological features of apoptosis in HeLa cells including the formation of apoptotic body-like vesicles, chromatin condensation and margination were observed by using 4, 6-diamidino-2- phenylindole dihydrochloride (DAPI) staining and fluorescence analysis. An apoptotic DNA ladder from 180 to 300 bp was also detected. Furthermore, 10 variation proteins associated with apoptosis pathway, viz. G3PDH isoforms 1/2 (G3PDH1/2), aldosereductase, ectodemal dysplasia receptor associated death receptor domain isoform CRA_a (EDARADD), heat shock 60kD protein 1 variant 1 (HSP60), heat shock 70kDa protein 5 precursor (HSP70), heat shock protein 90kDa beta member 1 precursor (HSP90), 14-3-3 protein ζ/δ, Ran and ubiquitin activating enzyme E1(UBE1), were identified from HMC-treated HeLa cells by the proteomic and quantitative real-time RT-PCR strategies. Importantly, the reactive oxygen species (ROS), mitochondrial membrane potential (Δψm) and caspase-9/3 activities were changed significantly in HMC-treated HeLa cells. Together, the data suggests that L. vannamei HMC mediates antiproliferative properties through the apoptosis mechanism involving the mitochondria triggered pathway.
Highlights
Hemocyanin (HMC) is an extracellular giant copper-containing glycoprotein found in the hemolymph of both mollusk and arthropod
Sulforhodamine B (SRB) assay was used to characterize the antiproliferative effect of HMC on HeLa cells viability
These data suggested that shrimp HMC exhibited antiproliferative effect on HeLa cells in vitro as an antitumor agent
Summary
Hemocyanin (HMC) is an extracellular giant copper-containing glycoprotein found in the hemolymph of both mollusk and arthropod. HMCs isolated from marine gastropods Rapana thomasiana and Megathura crenulata were doucumented to be acted as a potential bio-adjuvant for subunit vaccines [15]. Accumulating evidences indicate that HMCs from some mollusks have antitumor effects. Antonova et al found HMCs derived from two snail species: Helix lucorum (HlH) and Helix aspersa (HaH) possessed antitumor effects on multiple malignant cell lines including bladder cancer (CAL-29 and T-24), ovarian cancer (FraWü), acute monocytic leukemia (THP-1), prostate cancer (DU145), glioma cancer (LN-18), and Burkitt's lymphoma (Daudi) [25]. Arancibia et al documented that HMC isolated from gastropod Fissurella latimarginata (FLH) or Concholepas concholepas hemocyanin (CCH) could act as an effective antiproliferative agent and decrease tumor growth [27, 28]. The antiproliferative properties of HMC from shrimp L. vannamei against HeLa cells in vitro were investigated. Our data will assist in the investigation of multifunctionality of HMC and help to establish a potential strategy for cancer control
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call