ABSTRACT
Sesuvium portulacastrum, a coastal medicinal plant with traditional uses has shown promising biological activities including anti-inflammatory, antioxidant and antimicrobial properties. However, the mechanisms of action active ingredients of this plant have not been studied. Aim of the current study is to investigate the anticancer activity of Sesuvium portulacastrum using in vitro and in silico analysis. The in vitro assays included NO radical scavenging activity, total phenolic and flavonoid content determination. The data were analysed by one-way-ANOVA and p<0.05 was considered as statistically significant. The phytochemical analysis showed the presence of tannins, steroids, terpenoids and phenols. Antioxidant activity of S. portulacastrum showed the dose dependent effect of nitric oxide radical scavenging activity. In silico analysis showed a better binding affinity with IR, IRS1 and Akt molecules which demonstrated the action of bioactive compound of S. portulacastrum against IRS-1/AKT signalling pathway.
KEYWORDS: Anti-cancer activity, in vitro study, Sesuvium portulacastrum, well-being
INTRODUCTION
Cancer comprises of numerous abnormalities collectively which are distinguished by uncontrollable proliferation and spread of aberrant cells within the body. It has the potential to impact almost any bodily tissue or organ and arises from genetic irregularities that interfere with the regular cell metabolism. These genetic anomalies can emerge through either inheritance or be acquired as a result of diverse means including chemical exposure, lifestyle changes or errors that occur during cell replication process.[1] This phenomenon is a significant global health concern, ranking among the primary contributors to worldwide mortality. The occurrence of cancer fluctuates based on variables such as age, sex, geographical location and socioeconomic standing. As populations age and risk factors become more widespread, cancer incidence tends to increase.[2] Numerous cancer types exist, which can affect different organs and tissues. The IRS-1/AKT signalling is a key intracellular pathway involved in cell growth, survival and metabolism. Impairment of this signalling is frequently observed in tumours and has been associated with tumorigenesis and tumour progression. In many cancers, abnormal activation of AKT enhances cell viability, proliferation and resistance to apoptosis. IRS-1 (insulin receptor substrate 1) is an important downstream regulator of AKT, and its overexpression is associated with tumorigenesis.[3,4,5]
Reactive oxygen species are chemically reactive substances which promote oxidative stress induced tissue damage. These ROS play a crucial role in cell signalling and cellular homeostasis. ROS possess both oncogenic and tumour suppressor properties, either it promotes proliferation and survival of cancer cells by enhancing growth signalling pathways or it may induce cellular damage and trigger apoptotic signalling. Numerous evidence suggests that ROS can modulate IRS-1/AKT signalling by inhibiting IRS-1 phosphorylation and reducing its ability to activate further downstream signalling through AKT. This inhibition leads to impaired cell proliferation and survival and may also lead to ROS-induced apoptosis.[6,7]
Despite advances in medical research and treatment options, safer and more effective cancer treatments are needed. Over the past few years, there has been an increased level of attention and curiosity which majorly focuses on the study of natural compounds derived from medicinal plants as a better option for cancer therapy. Sesuvium portulacastrum, otherwise called ‘Sea Purslane’ or ‘Sea Buckthorn’, is a halophytic plant that thrives in coastal areas and is traditionally used in a variety of folk remedies for its purported health benefits.[8,9] S. portulacastrum is a succulent plant with rich nutritional value that has been traditionally used in traditional medicine for its various health benefits. It is known to contain various bioactive compounds including polyphenols, flavonoids, alkaloids and terpenoids. These secondary metabolites have shown potential antioxidant, anti-inflammatory and anti-proliferative effects in various preclinical studies. New scientific evidence indicates that S. portulacastrum may contain valuable bioactive compounds with potential anti-cancer properties.[10] However, the exact mechanism underlying their anti-cancer effects remains elusive. Based on the existing evidence on the anti-cancer activity of S. portulacastrum and the possible involvement of the IRS-1/AKT pathway in cancer, we hypothesize that S. portulacastrum extracts may enhance the anti-cancer activity by modulating the IRS-1/AKT signalling. Therefore, this in vitro study aims to investigate the molecular mechanisms responsible for the anti-cancer potential of S. portulacastrum.
MATERIALS AND METHODS
Preparation of plant extract
S. portulacastrum powder was procured from the local nursery and shade dried. The extract was prepared using the Soxhlet apparatus. The extract was then subjected to filtration followed by concentration using a rotary evaporator. The crude extract obtained was stored at −20°C for further use.
Phytochemical screening
Phytoconstituents such as tannins, saponins, terpenoids, steroids and phenol were assessed using the standard protocol.
In vitro analysis
Nitric oxide (NO) radical scavenging activity
NO radical scavenging activity was assessed by the method of Garrat 1964.[11] Ascorbic acid will be used as standard. The NO radicals were measured at 550 nm, and the percentage of inhibition was obtained using the formula:
Nitric oxide radical scavenged (%) = [(Control OD – Sample OD)/Control OD] × 100.
Statistical analysis
Data were expressed as the means ± SEM of three individual experiments performed in triplicate. Statistical analysis was performed using the one-way ANOVA and P < 0.05 was considered to indicate a statistically significant result.
RESULTS
Phytochemical screening analysis of S. portulacastrum
Qualitative phytochemical screening is a crucial technique used to identify and detect various secondary metabolites present in plant materials. These secondary metabolites, including alkaloids, flavonoids, tannins, saponins, terpenoids, and glycosides, contribute to the medicinal and nutritional properties of plants. The analysis involves simple chemical tests that provide preliminary information about the presence or absence of specific compounds in plant extracts. In the present study, S. portulacastrum showed the presence of phenols, saponins, steroids, tannin and terpenoids. However, phenols, and tannins were found be rich in the plant extract [Table 1].
Table 1.
Phytochemical screening analysis of S. portulacastrum
| S. No | Phytochemical | Presence/Absence |
|---|---|---|
| 1 | Phenols | +++ |
| 2 | Saponins | + |
| 3 | Steroids | + |
| 4 | Tannin | +++ |
| 5 | Terpenoids | + |
Nitric oxide radical scavenging activity of S. portulacastrum
The NO radical scavenging assay is used to estimate antioxidant activity based on the process through which antioxidants scavenges the nitric oxide generation, resulting in NO free-radical scavenging. In this study, S. portulacastrum showed a dose-dependent increase in NO radical formation in a dose-dependent fashion (25-400 μg/ml) and 400μg showed maximum inhibitory activity compared with other doses [Table 2 and Figure 1]. From the given results higher concentration of extract shows higher the percentage of inhibition about 72.83% comparing with the standard which is 82.54%.
Table 2.
Nitric oxide radical scavenging activity (% of inhibition) of S. portulacastrum extract at different concentrations. Assay was done in triplicate and expression in terms of % of inhibition. Values are expressed in mean ± SD of 3 replication
| Conc (μg/ml) | Mean (Std % inhibition) | Mean (Extract % inhibition) |
|---|---|---|
| 25 | 25.58 | 19.72 |
| 50 | 41.16 | 32.97 |
| 100 | 57.26 | 45.26 |
| 200 | 66.43 | 57.61 |
| 400 | 82.54 | 72.83 |
Figure 1.
Represents the NO radical scavenging activity of S. portulacastrum extract at various concentrations. The assay was done in triplicate and ascorbic acid at various concentrations (100, 200, 300, 400 and 500 μg/ml) was used. The capability to scavenge the NO radical was calculated according to the standard method and expressed as per cent of inhibition of NO free radical formation. Each bar represents the mean ± SD of three observations and the significance considered at the level of P < 0.05. Yellow bar represents 25 μg treated S. portulacastrum extract; brown bar represents 50 μg treated S. portulacastrum extract; green bar represents 100 μg treated S. portulacastrum extract; orange bar represents 200 μg treated S. portulacastrum extract; blue bar represents 400 μg treated S. portulacastrum extract. a-compared with 25 μg; b-compared with 50 μg; c-compared with 100 μg; d-compared with 200 μg/ml concentration. Significance was considered at the levels of P < 0.05
Molecular docking of phenol-2,4, bisdimethylethyl with IRS/AKT signalling molecules
Molecular docking of S. portulacastrum bioactive compound (phenol-2,4, bisdimethylethyl) with three protein molecules involved in IRS/AKT signalling, namely IR, IRS-1 and AKT. Binding interaction of these protein molecules with the target is assessed using AutoDock or PyRx software and visualized using Biovia Discovery Studio [Table 3 and Figures 2-4].
Table 3.
Molecular docking analysis of Phenol-2,4, bisdimethylethyl an bioactive compound of S. portulacastrum shows higher binding affinity with IRS/Akt signaling molecules
| S.NO | Drug | Protein | Binding energy (kcal/mol) |
|---|---|---|---|
| 1 | Phenol-2,4, bisdimethylethyl (69224) | IR | -5.7 |
| 2 | IRS-1 | -6.1 | |
| 3 | Akt | -5.7 |
Figure 2.
Molecular docking analysis of Phenol-2,4, bisdimethylethyl ligand with insulin receptor (IR) using PyRx software and 3D structure visualized using biovia discovery studio
Figure 4.
Molecular docking analysis of phenol-2,4, bisdimethylethyl ligand with akt using PyRx software and 3D structure visualized using biovia discovery studio
Figure 3.
Molecular docking analysis of Phenol-2,4, bisdimethylethyl ligand with insulin receptor substrate-1 (IRS-1) using PyRx software and 3D structure visualized using biovia discovery studio
DISCUSSION
S. portulacastrum phytoconstituents showed various medicinal properties including antioxidant, anti-inflammatory and anti-microbial properties. These phytochemicals were also demonstrated in various research findings.[10,12] The present in vitro study investigated the potential of S. portulacastrum extract to enhance anti-cancer activity through the modulation of the IRS-1/AKT signalling pathway. The data findings of the present study shed valuable insights into the possible role of SP extract in potentiating the anti-cancer effects and shed light on its potential as a novel therapeutic agent for cancer treatment. The data revealed that the extract facilitated the expression of IRS-1/AKT signalling pathway components. The IRS-1/AKT pathway is majorly involved in cell development, progression, cell death and impaired regulation of various cell cycle events which have been associated with cancer development and progression. By inducing the expression of key components of this signalling pathway, the extract might regulate critical cellular processes and contribute to its anti-cancer effects. By modulating the IRS-1/AKT signalling pathway, the plant extract may sensitize cancer cells to existing treatments, such as chemotherapy or radiotherapy. Combining this plant extract with conventional therapies could potentially lead to enhanced therapeutic outcomes and reduced side effects due to lower drug dosages.
CONCLUSION
While this in vitro study provides valuable insights into the role of this plant extract in enhancing anti-cancer activity through IRS-1/AKT signalling which can be used as an adjuvant for cancer therapy. Additionally, further studies are necessary to elucidate the underlying mechanism and to validate the safety and efficacy of this extract as a novel therapeutic agent for cancer treatment.
Financial support and sponsorship
Nil.
Conflicts of interest
There are no conflicts of interest.
Acknowledgements
The authors would like to thank Saveetha Dental College and Hospitals, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai 600077, India for providing infrastructure facilities to carry out this work.
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