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Hevea brasiliensis latex dialysed C-serum precipitate subfraction exerts a negligible level of genotoxicity in the Ames test, mouse lymphoma assay and micronucleus assay

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Abstract

Dialysed C-serum precipitate (DCP), a sub-fraction of latex C-serum originating from Hevea brasiliensis, has shown specific in vitro antiproliferative properties towards cancer-origin cell lines. As a potential agent to be used in cancer treatment, preclinical safety assessment tests are crucial. This study focused on investigating the mutagenicity of DCP via bacterial reverse mutation assay (Ames test), Mouse lymphoma assay (MLA), and micronucleus assay according to OECD guidelines. No mutagenicity was exhibited by DCP at concentrations lower than 1 mg/mL for the five different auxotroph strains of Salmonella typhimurium (TA 98, TA 100, TA 1535, and TA 1537) and Escherichia coli WP2uvrA pKM101 in the absence and presence of metabolic activation system (S9-mix). Nonetheless, it has been found that DCP was able to induce negligible mutation in MLA and micronucleus assay. Overall, DCP does express dose-dependant mutagenicity according to the assays conducted and thus, it is crucial to take this into consideration should DCP be used in the development of therapeutic agent.

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Data availability

The data that support the findings of this study are not openly available due to reasons of sensitivity and are available from the corresponding author upon reasonable request. Data are located in controlled access data storage at Instutute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800 USM, Pulau Pinang, Malaysia.

Abbreviations

2-AA:

2-Aminoanthracene

2-NF:

2-Nitrofluorene

9-AA:

9-Aminoacridine

BN:

Binucleated

BP:

Benzopyrene

CBPI:

Cytokinesis block proliferation index

CB:

Cytochalasin B

DCP:

Dialysed C-serum precipitate

dThd:

Thymidine deoxyriboside

dTMP:

Deoxythymidylate

E.coli :

Escherichia coli

GEF:

Global evaluation factor

HS:

Horse serum

IMF:

Induced mutant frequency

KCl:

Potassium chloride

LC:

Large colony

MF:

Mutant frequency

MI:

Mutagenicity index

MLA:

Mouse lyphoma assay

MMS:

Methylmethane sulfonate

MN:

Micronuclues

NaN3 :

Sodium azide

NCE:

New chemical entities

NDA:

New drug application

OECD:

Organisation for economic co-operation and development

PBS:

Phosphate buffered saline

R10:

RPMI 1640 with 10% HS

R20:

RPMI 1640 with 20% HS

R5:

RPMI 1640 with 5% HS

RPD:

Relative population index

RPMI 1640:

Roswell Park Memorial Institute Media

RSG:

Relative suspension growth

RTG:

Relative total growth

RV:

Relative viability

S. typhimurium :

Salmonella typhimurium

SC:

Small colony

SMF:

Spontaneous mutant frequency

TFT:

Triflourothymidine

THG:

R10 media supplemented with thymidine, hypoxanthine and glycine

THMG:

R10 media supplemented with thymidine, hypoxanthine, methotrexate and glycine

Tk :

Thymidine kinase

TX:

Triton X-100

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Funding

This study was supported by the Malaysia Ministry of Higher Education under the Fundamental Research Grant Scheme (FRGS/1/2014/SG03/USM/02/1) and the Malaysian Rubber Board.

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Authors and Affiliations

  1. Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Jalan Inovasi, 11800 USM, Pulau Pinang, Malaysia

    Malini Devi Raman & Ming Thong Ong

  2. Rubber Research Institute of Malaysia, Malaysian Rubber Board, P.O. Box 10150, Kuala Lumpur, Malaysia

    Norazreen Abd Rahman & Sunderasan Elumalai

  3. School of Pharmaceutical Sciences, Universiti Sains Malaysia, 11800 USM, Pulau Pinang, Malaysia

    Vikneswaran Murugaiyah

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  1. Malini Devi Raman
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Correspondence to Ming Thong Ong.

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Raman, M.D., Abd Rahman, N., Elumalai, S. et al. Hevea brasiliensis latex dialysed C-serum precipitate subfraction exerts a negligible level of genotoxicity in the Ames test, mouse lymphoma assay and micronucleus assay. J Rubber Res 26, 139–153 (2023). https://doi.org/10.1007/s42464-023-00201-x

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