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Investigational New Drugs
Published in: Investigational New Drugs 2/2021

01-04-2021 | Acute Lymphoblastic Leukemia | PRECLINICAL STUDIES

Recombinant human lactoferrin carrying humanized glycosylation exhibits antileukemia selective cytotoxicity, microfilament disruption, cell cycle arrest, and apoptosis activities

Authors: Sayuri Nakamura-Bencomo, Denisse A. Gutierrez, Elisa Robles-Escajeda, Blanca Iglesias-Figueroa, Tania S. Siqueiros-Cendón, Edward A. Espinoza-Sánchez, Sigifredo Arévalo-Gallegos, Renato J. Aguilera, Quintín Rascón-Cruz, Armando Varela-Ramirez

Published in: Investigational New Drugs | Issue 2/2021

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Summary

Lactoferrin has gained extensive attention due to its ample biological properties. In this study, recombinant human lactoferrin carrying humanized glycosylation (rhLf-h-glycan) expressed in the yeast Pichia pastoris SuperMan5, which is genetically glycoengineered to efficiently produce functional humanized glycoproteins inclosing (Man)5(GlcNAc)2 Asn-linked glycans, was analyzed, inspecting its potential toxicity against cancer cells. The live-cell differential nuclear staining assay was used to quantify the rhLf-h-glycan cytotoxicity, which was examined in four human cell lines: acute lymphoblastic leukemia (ALL) CCRF-CEM, T-cell lymphoblastic lymphoma SUP-T1, cervical adenocarcinoma HeLa, and as control, non-cancerous Hs27 cells. The defined CC50 values of rhLf-h-glycan in CCRF-CEM, SUP-T1, HeLa, and Hs27 cells were 144.45 ± 4.44, 548.47 ± 64.41, 350 ± 14.82, and 3359.07 ± 164 µg/mL, respectively. The rhLf-h-glycan exhibited a favorable selective cytotoxicity index (SCI), preferentially killing cancer cells: 23.25 for CCRF-CEM, 9.59 for HeLa, and 6.12 for SUP-T1, as compared with Hs27 cells. Also, rhLf-h-glycan showed significant antiproliferative activity (P < 0.0001) at 24, 48, and 72 h of incubation on CCRF-CEM cells. Additionally, it was observed via fluorescent staining and confocal microscopy that rhLf-h-glycan elicited apoptosis-associated morphological changes, such as blebbing, nuclear fragmentation, chromatin condensation, and apoptotic bodies in ALL cells. Furthermore, rhLf-h-glycan-treated HeLa cells revealed shrinkage of the microfilament structures, generating a speckled/punctuated pattern and also caused PARP-1 cleavage, a hallmark of apoptosis. Moreover, in ALL cells, rhLf-h-glycan altered cell cycle progression inducing the G2/M phase arrest, and caused apoptotic DNA fragmentation. Overall, our findings revealed that rhLf-h-glycan has potential as an anticancer agent and therefore deserves further in vivo evaluation.
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Metadata
Title
Recombinant human lactoferrin carrying humanized glycosylation exhibits antileukemia selective cytotoxicity, microfilament disruption, cell cycle arrest, and apoptosis activities
Authors
Sayuri Nakamura-Bencomo
Denisse A. Gutierrez
Elisa Robles-Escajeda
Blanca Iglesias-Figueroa
Tania S. Siqueiros-Cendón
Edward A. Espinoza-Sánchez
Sigifredo Arévalo-Gallegos
Renato J. Aguilera
Quintín Rascón-Cruz
Armando Varela-Ramirez
Publication date
01-04-2021
Publisher
Springer US
Published in
Investigational New Drugs / Issue 2/2021
Print ISSN: 0167-6997
Electronic ISSN: 1573-0646
DOI
https://doi.org/10.1007/s10637-020-01020-2

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