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. 2022 Jul 30;11(15):1994.
doi: 10.3390/plants11151994.

Antioxidant and Anticancer Potential of Bioactive Compounds from Rhinacanthus nasutus Cell Suspension Culture

Pattralak Songserm  1   2 , Poramaporn Klanrit  3 , Poramate Klanrit  4   5 , Jutarop Phetcharaburanin  4   5 , Pornthap Thanonkeo  2   6 , Jirawan Apiraksakorn  2   6 , Khamphee Phomphrai  7 , Preekamol Klanrit  2   5   6
Affiliations

Antioxidant and Anticancer Potential of Bioactive Compounds from Rhinacanthus nasutus Cell Suspension Culture

Pattralak Songserm et al. Plants (Basel). .

Abstract

The potential benefits of natural plant extracts have received attention in recent years, encouraging the development of natural products that effectively treat various diseases. This is the first report on establishing callus and cell suspension cultures of Rhinacanthus nasutus (L.) Kurz. A yellow friable callus was successfully induced from in vitro leaf explants on Murashige and Skoog medium supplemented with 1 mg/L 2,4-dichlorophenoxyacetic acid and 1 mg/L 1-naphthalene acetic acid. A selected friable callus line was used to establish the cell suspension culture with the same medium. The antioxidant assays showed that the leaf- and ethanolic-suspension-cultured cell (SCC) extracts exhibited high antioxidant potential. In addition, the in vitro cytotoxicity revealed by the MTT assay demonstrated potent antiproliferative effects against the oral cancer cell lines ORL-48 and ORL-136 in a dose-dependent manner. Several groups of compounds, including terpenoids, phenolics, flavonoids, quinones, and stilbenes, were identified by UHPLC-QToF-MS, with the same compounds detected in leaf and SCC extracts, including austroinulin, lucidenic acid, esculetin, embelin, and quercetin 3-(2″-p-hydroxybenzoyl-4″-p-coumarylrhamnoside). The present study suggests the value of further investigations for phytochemical production using R. nasutus cell suspension culture.

Keywords: Rhinacanthus nasutus; anticancer; antioxidant; callus; cell suspension culture.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Rhinacanthus nasutus (L.) Kurz—A Thai medicinal plant used in the present study: (A) a mature plant, (B) a shoot tip, and (C) a flower.
Figure 2
Figure 2
Callus culture of R. nasutus: (A) callus initiation from leaf explants after 3 weeks; (B) callus growth on CIM (MS medium supplemented with 1 mg/L 2,4-D and 1 mg/L NAA) after culturing for 7 days; (C) callus proliferation on CIM after 15 days of culture.
Figure 3
Figure 3
Establishment of R. nasutus cell suspension culture: (A) leaf-derived yellow friable callus; (B) SCCs of R. nasutus; (C) morphology of 15 day friable callus under a bright-field microscope. The scale bar represents 25 µm.
Figure 4
Figure 4
Growth profile of R. nasutus cell suspension cultures during 30 days of culture in liquid CIM.
Figure 5
Figure 5
Antioxidant capacity of the ethanolic leaf and SCC extracts of R. nasutus (A) as determined by FRAP assay and (B) by DPPH and ABTS assays. The data are presented as the means ± SD of the results of triplicate determinations. Bars with different letters are significantly different according to an independent-samples t test at p ≤ 0.05.
Figure 6
Figure 6
Oral cancer cell viability after treatment with ethanolic leaf and SCC extracts of R. nasutus: (A) ORL-48 cell line and (B) ORL-136 cell line. The data are presented as the means ± SD of the results of triplicate determinations. Bars with different letters at each concentration are significantly different according to an independent-samples t test at p ≤ 0.05.
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