Antimutagenic and Chemopreventive Properties of 6-(Methylsulfinyl) Hexyl Isothiocyanate on TK6 Human Cells by Flow Cytometry

Abstract

6-(methylsulfinyl) hexyl isothiocyanate (6-MITC), is the main bioactive compound present in Wasabia japonica rhizome. Several scientific studies have shown that 6-MITC possesses interesting antimicrobial, anti-inflammatory, antiplatelet and antioxidant properties which therefore suggested us it could have an interesting chemopreventive potential. In a recent publication, we demonstrated, in two different leukemia cell lines, its ability to modulate several mechanisms supporting its antitumor activity. For this reason, we thought useful to continue the research, by investigating the potential antimutagenic activity of 6-MITC and thus better define its profile as a possible chemopreventive agent. 6-MITC antimutagenic effect against two known mutagenic agents: the clastogen Mitomycin C (MMC) and the aneuplodogen Vinblastine (VINB), was analyzed, in terms of micronuclei frequency decrease, after short-and long-time treatment on TK6 human cells, using a new automated protocol of the "In Vitro Mammalian Cell Micronucleous Test" by flow cytometry. The results showed a different behavior of the isothiocyante. In particular, 6-MITC was unable to counteract the MMC genotoxicity, but when it was associated with VINB a statistically significant decrease in the micronuclei frequency was registered. Overall, the results obtained suggest a potential antimutagenic activity of 6-MITC, in particular against the aneuploidogen agents. This ability, to inhibit or counteract the mutations at the cellular level has a great therapeutic value and it represents a mechanism through a chemopreventive agent can express its activity.

6-(Methylsulfonyl) Hexyl Isothiocyanate: A Chemopreventive Agent Inducing Autophagy in Leukemia Cell Lines

“…Research previously conducted in our laboratories proved the capacity of this ITC to interact with different cellular and molecular targets critical in neoplastic development, supporting its antitumoral activity [ 52 , 53 ]. In particular, in two other leukemia cell lines (Jurkat and HL-60), 6-MITC has been demonstrated to be capable of cytotoxic, cytostatic, and cytodifferentiation effects.…”

“…Still, on the other hand, it displayed a potential antigenotoxic activity against the aneuploidogen Vinblastine, which acts on cellular mitosis by preventing tubulin polymerization and consequently inhibits the microtubule aggregation [ 64 ]. This result seems to suggest that the ITC tested cannot counteract a structural DNA damage but that it can intervene in the mitotic spindle formation or at the chromosomal segregation time [ 53 ].…”

“…In addition, in two recent publications, we have proved that 6-MITC elicits cytotoxic and cytostatic effects, induces cell differentiation on the leukemic cell lines Jurkat and HL-60, and exerts antimutagenic activity on TK6 cells [ 52 , 53 ]. Overall, these results demonstrated the capacity of 6-MITC to modulate several mechanisms supporting its antitumor activity.…”

Anthraquinones: Genotoxic until Proven Otherwise? A Study on a Substance-Based Medical Device to Implement Available Data for a Correct Risk Assessment

“…In 2018, we published an innovative flow cytometric protocol to score MNi to overcome some critical issues affecting the classic method of analysis by optical microscopy, such as the subjectivity of interpretation, long samples preparation and analysis times and the number of events examined, which is extremely low for the purpose of a robust statistical analysis. Since then, our protocol was successfully used to prove the mutagenicity of different novel psychoactive substances [ 20 , 21 , 22 ] and the lack of mutagenic capacity of Wasabia japonica [ 19 ].…”

“…OECD guideline n° 487 lists different possible cell lines suitable to analyse the presence of MNi (e.g., CHO, V79, CHL/IU, L5178Y or TK6), among which the most used are TK6 cells, by virtue of their human and non-tumoral origin, ease of maintenance in culture and replicative speed [ 16 , 18 ]. Moreover, this cell line grows in suspension and is therefore an optimal assay system for flow cytometric analyses [ 19 , 20 , 21 , 22 ].…”