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Different approaches have been evaluated to reach this objective through the development of OLE and HT derivatives or carrier systems.
|
[] |
ChemBL_V1
|
For example, peracetylated OLE had a stronger anti-proliferative and antioxidant activity than OLE in thyroid and breast cancer in vitro ; iron oxide nanoparticles coated with glucose and conjugated with OLE showed a notable cytotoxic action against colorectal cancer cell line SW480 ; chitosan nanoencapsulation of HT was shown to be unaffected by time-dependent dynamic changes in efficacy observed for free HT in lung and breast cancer cell lines ; hydroxytyrosyl dodecyl ether, an HT alkyl ether derivative, had a very strong cytotoxic activity at low concentrations (IC50 = 19.9 ± 4.6 μM) in lung cancer cells in vitro ; hydroxytyrosyl oleate, an HT ester, exerted the same effects shown by HT on SH-SY5Y neuroblastoma cells at lower concentrations ; HT-loaded poly lactide-co-glycolide-co-polyacrylic acidnanoparticles exhibited cytotoxic and transcriptional regulating effects at lower concentrations (6 ppm) than free HT in colorectal cancer in vitro .
|
[
{
"end": 283,
"label": "CellLine",
"start": 278,
"text": "SW480"
},
{
"end": 709,
"label": "CellLine",
"start": 702,
"text": "SH-SY5Y"
}
] |
ChemBL_V1
|
Anyway, it would be highly recommendable to address future OLE- and HT-specific research directions towards the exploration of the true effect of nutritionally relevant concentrations of these phenols (or OLE and HT concentrations that can realistically be maintained in plasma) on cancer cells, since even most recent evidence is not able to determine if the administration of OLE and HT in humans could be detrimental in terms of tumor growth and anti-cancer drug metabolism/effects.
|
[] |
ChemBL_V1
|
Up to now, conclusive studies assessing the influence of nutritionally relevant concentrations of OLE and HT on tumor insurgence and expansion, as well as proof of the absence of OLE- and HT-mediated alterations of chemotherapy metabolism, are still missing.
|
[] |
ChemBL_V1
|
Another important reason for concern is represented by OLE and HT context-dependent dual action as both antioxidant and pro-oxidant agents, since such a behavior may alter the efficacy of cancer treatment in unexplored ways.
|
[] |
ChemBL_V1
|
Also, OLE and HT modulatory activity on immune cells deserves further investigation.
|
[] |
ChemBL_V1
|
Currently, neither OLE nor HT could be safely considered as cancer-preventive agents or drugs for combinatory therapies without excluding the possibility that nutritionally relevant concentrations of these compounds might facilitate neoplastic cell expansion or even treatment escape.
|
[] |
ChemBL_V1
|
The ayurvedic plant Withania somnifera, a member of the Solanaceae family, has been used as a remedy for diverse health problems, including cancer.
|
[] |
ChemBL_V1
|
The objective of this investigation was to conduct a comparative analysis of the in vitro cytotoxic properties of methanolic extracts derived from the leaf, stem, and root of W. somnifera on HepG2 and L929 cell lines.
|
[
{
"end": 196,
"label": "CellLine",
"start": 191,
"text": "HepG2"
},
{
"end": 205,
"label": "CellLine",
"start": 201,
"text": "L929"
}
] |
ChemBL_V1
|
Methanolic extracts were obtained using the Soxhlet extraction method.
|
[] |
ChemBL_V1
|
To assess the in vitro anticancer action on the HepG2 and L929 cell lines, an MTT assay was performed.
|
[
{
"end": 53,
"label": "CellLine",
"start": 48,
"text": "HepG2"
},
{
"end": 62,
"label": "CellLine",
"start": 58,
"text": "L929"
}
] |
ChemBL_V1
|
Changes in cell morphology were observed using an inverted microscope.
|
[] |
ChemBL_V1
|
The MTT assay results indicated that the leaf, stem, and root methanolic extracts of W. somnifera showed significantly higher in vitro cytotoxicity in HepG2 cells, with IC50 values of 43.06 ± 0.615, 45.60 ± 0.3, and 314.4 ± 0.795 μg/mL than in L929 cell lines with 78.77 ± 0.795, 90.55 ± 0.800, and 361.70 ± 0.795 μg/mL, respectively.
|
[
{
"end": 156,
"label": "CellLine",
"start": 151,
"text": "HepG2"
},
{
"end": 248,
"label": "CellLine",
"start": 244,
"text": "L929"
}
] |
ChemBL_V1
|
The leaf methanolic extract was the most effective, followed by the stem methanolic extract in the HepG2 cell line.
|
[
{
"end": 104,
"label": "CellLine",
"start": 99,
"text": "HepG2"
}
] |
ChemBL_V1
|
The results of our study have confirmed that the methanolic extracts of both the leaf and stem of W. somnifera exhibit significant in vitro cytotoxicity in HepG2 cell lines, while displaying no significant cytotoxicity in the L929 cell line.
|
[
{
"end": 161,
"label": "CellLine",
"start": 156,
"text": "HepG2"
},
{
"end": 230,
"label": "CellLine",
"start": 226,
"text": "L929"
}
] |
ChemBL_V1
|
Furthermore, the data obtained from the MTT assay indicate that the leaf methanolic extract possesses a more potent cytotoxic activity than the stem methanolic extract with respect to the HepG2 cell line.
|
[
{
"end": 193,
"label": "CellLine",
"start": 188,
"text": "HepG2"
}
] |
ChemBL_V1
|
Further studies on the identification and isolation of bioactive metabolites are required to explore the mechanisms underlying their in vitro cytotoxicity.
|
[] |
ChemBL_V1
|
Ayurvedic medicines have long been used with various medicinal plants and their parts in several medicinal formulations .
|
[] |
ChemBL_V1
|
Withania somnifera is an important herb that belongs to the Solanaceae family.
|
[] |
ChemBL_V1
|
It is also known as Indian ginseng, winter cherry, or ashwagandha .
|
[] |
ChemBL_V1
|
Indians have been using this herb for therapeutic purposes since time immemorial .
|
[] |
ChemBL_V1
|
However, systemic scientific research on this plant began in the 1950s only .
|
[] |
ChemBL_V1
|
Initially, it was used to treat fertility and reproductive health issues, but it is now used to prevent aging, boost important body fluids such as blood cells, lymph secretion, semen, and nourish various body organs .
|
[] |
ChemBL_V1
|
Despite advancements in medicine, cancer remains the main cause of mortality worldwide .
|
[] |
ChemBL_V1
|
Among noncommunicable diseases, cancer is the second leading cause of death .
|
[] |
ChemBL_V1
|
Uncontrolled cell proliferation, metastasis, invasion, programmed cell death, and angiogenesis are hallmarks.
|
[] |
ChemBL_V1
|
Cancer can affect any organ in the human body, although it most frequently affects the breast, lung, liver, colon, prostate, kidney, and ovaries .
|
[] |
ChemBL_V1
|
Approximately 29% of all malignancies are lung cancers .
|
[] |
ChemBL_V1
|
Breast cancer is the major cause of morbidity in women, whereas prostate cancer is the major cause of morbidity in men.
|
[] |
ChemBL_V1
|
Liver cancer ranks third in terms of cancer mortality .
|
[] |
ChemBL_V1
|
The rate of death from cancer is constantly increasing .
|
[] |
ChemBL_V1
|
The lack of efficient anticancer drugs is a major burden on the healthcare system, and there is an urgent need for such drugs .
|
[] |
ChemBL_V1
|
Despite the availability of many chemoprotective medications for cancer treatment, these are expensive and have numerous side effects .
|
[] |
ChemBL_V1
|
Therefore, it is critically important to search for cost-effective and promising natural medications with minimal side effects to reduce cancer morbidity rates.
|
[] |
ChemBL_V1
|
Herbal medicines are considered the most viable ways to cure cancer .
|
[] |
ChemBL_V1
|
A variety of plants, such as W. somnifera, are used for drug development.
|
[] |
ChemBL_V1
|
These are more effective and have fewer adverse effects.
|
[] |
ChemBL_V1
|
Thus, there is a pressing demand for natural medications to stop cancer progression and spread throughout the body .
|
[] |
ChemBL_V1
|
W. somnifera has received a lot of attention recently for its anticancer studies .
|
[] |
ChemBL_V1
|
W. somnifera extracts have been investigated from various plant parts for therapeutic value in treating cancers of various origins .
|
[] |
ChemBL_V1
|
In one study, a decrease in mammary carcinomas in mice was observed with W. somnifera root extract .
|
[] |
ChemBL_V1
|
In another study, human umbilical vein endothelial cells (HUVECs) were treated with W. somnifera root extract, and withaferin A reduced cell proliferation .
|
[] |
ChemBL_V1
|
W. somnifera leaf extracts have been employed as a treatment during research investigations exploring its potential anticancer effects .
|
[] |
ChemBL_V1
|
Previous studies have shown that W. somnifera extracts reduce the proliferation of MCF-7, pancreatic, prostate, kidney, and fibrosarcoma cells .
|
[
{
"end": 88,
"label": "CellLine",
"start": 83,
"text": "MCF-7"
}
] |
ChemBL_V1
|
Consequently, it can be concluded that substances extracted from W. somnifera have strong antineoplastic activity and could potentially be used as chemotherapeutic agents.
|
[] |
ChemBL_V1
|
Moreover, several investigations have demonstrated the anticancer activities of W. somnifera during the course of research on the anticancer activities of this plant for the development of herbal-based drugs .
|
[] |
ChemBL_V1
|
The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric method is a rapid and effective assay for assessing cellular metabolic activity, cytotoxicity, and proliferation .
|
[] |
ChemBL_V1
|
In light of the comprehensive assessments documenting the anticancer attributes of W. somnifera , this article presents in vitro comparative investigations into the anticancer potentials of distinct constituents of W. somnifera, specifically the leaf, stem, and root, in the selected cell lines HepG2 and L929.
|
[
{
"end": 300,
"label": "CellLine",
"start": 295,
"text": "HepG2"
},
{
"end": 309,
"label": "CellLine",
"start": 305,
"text": "L929"
}
] |
ChemBL_V1
|
All chemicals and media were procured from Hi-Media and Sigma-Aldrich, respectively.
|
[] |
ChemBL_V1
|
The human liver cancer cell line HepG2 and the mouse fibroblast noncancerous cell line L929 were acquired from the National Centre for Cell Sciences (NCCS), Pune.
|
[
{
"end": 38,
"label": "CellLine",
"start": 33,
"text": "HepG2"
},
{
"end": 91,
"label": "CellLine",
"start": 87,
"text": "L929"
}
] |
ChemBL_V1
|
The cell lines were later propagated in Dulbecco's Modified Eagle Medium (DMEM), supplemented with 10% (v/v) fetal bovine serum (FBS), an antimycotic antibiotic, and maintained with a continuous supply of 5% CO2 at 37°C.
|
[] |
ChemBL_V1
|
The W. somnifera seeds were bought from Zooqa Herbs, Chennai, Tamil Nadu.
|
[] |
ChemBL_V1
|
The seeds were sown and matured under natural soil and light conditions at the Department of Genetics and Biotechnology, Osmania University, Hyderabad.
|
[] |
ChemBL_V1
|
The plant materials were collected in the months of July–October 2022, during the monsoon season.
|
[] |
ChemBL_V1
|
The collected plant material was thoroughly washed three times with running water.
|
[] |
ChemBL_V1
|
Then, the plant material was rinsed once with sterile distilled water and weighted.
|
[] |
ChemBL_V1
|
The plant material was air dried in the shade at room temperature and weighed again.
|
[] |
ChemBL_V1
|
The fresh sample weight to dried sample weight ratios 37.02 : 23.82 of leaf, 28.72 : 16.92 of stem, and 28.31 : 14.71 of root, respectively, were used to coarsely grind and pulverise.
|
[] |
ChemBL_V1
|
The authentication of W. somnifera was carried out by Dr. A. Vijaya Bhaskar Reddy, Botany Department, Osmania University, Hyderabad.
|
[] |
ChemBL_V1
|
The plant was deposited in the herbarium of the Botany Department, Osmania University, Hyderabad, with voucher number GEN/OU/001-2018-HY.
|
[] |
ChemBL_V1
|
The leaf, stem, and root parts of W. somnifera were grounded coarsely and extracted with methanol in a Soxhlet for 24 hours and then air-dried.
|
[] |
ChemBL_V1
|
A viscous semisolid mass was produced using a rotary evaporator to concentrate the extract under reduced pressure at 40°C.
|
[] |
ChemBL_V1
|
HepG2 and L929 cells were grown in Dulbecco's modified Eagle's medium (DMEM) (Himedia) containing streptomycin (100 μg/mL) and penicillin (100 μg/mL), supplemented with 10% (v/v) FBS.
|
[
{
"end": 5,
"label": "CellLine",
"start": 0,
"text": "HepG2"
},
{
"end": 14,
"label": "CellLine",
"start": 10,
"text": "L929"
}
] |
ChemBL_V1
|
Cells were cultured at 37°C and 5% CO2; the complete medium was changed every three days (21, 22).
|
[] |
ChemBL_V1
|
In this study, cells in 6-well plates were used to examine the morphology of the cell lines in vitro under an inverted microscope.
|
[] |
ChemBL_V1
|
Cell viability and in vitro cytotoxicity were evaluated using an MTT assay [21–23].
|
[] |
ChemBL_V1
|
In order to ascertain the impact of the methanolic leaf, stem, and root extracts of W. somnifera on HepG2 and L929 cells, an MTT assay was conducted.
|
[
{
"end": 105,
"label": "CellLine",
"start": 100,
"text": "HepG2"
},
{
"end": 114,
"label": "CellLine",
"start": 110,
"text": "L929"
}
] |
ChemBL_V1
|
In 96-well plates, 100 μL of each cell line was seeded at a density of 10,000 cells per well.
|
[] |
ChemBL_V1
|
The plates were then incubated for 48 h at 37°C in a 5% CO2 environment.
|
[] |
ChemBL_V1
|
After incubation, the cells were examined in a half-confluent monolayer.
|
[] |
ChemBL_V1
|
Next, 20, 50, 100, 200, 500, 1000, 1500, and 2000 μg/mL of W. somnifera leaf, stem, and root methanolic extracts were treated in triplicate, as presented in Figure 1.
|
[] |
ChemBL_V1
|
The cells were then incubated at 37°C and 5% CO2 for 24 h. The untreated cell lines served as the negative control, and the cells treated with the anticancer drug doxorubicin served as the positive control.
|
[] |
ChemBL_V1
|
Cell lines treated with the extracts served as test samples.
|
[] |
ChemBL_V1
|
After 24 hours, the cells were observed using an inverted microscope to check for any morphological changes or cell death.
|
[] |
ChemBL_V1
|
After observation, the culture medium was removed, and 100 µL of fresh medium was added along with 10 µL of MTT reagent (5 mg/mL).
|
[] |
ChemBL_V1
|
The plates were then placed in an incubator with 5% CO2 at 37°C for 4 h. Subsequently, the medium containing MTT was removed, the formazan purple precipitate was solubilized, and 100 µL of DMSO was added.
|
[] |
ChemBL_V1
|
The plates were then incubated for 1 hour at 37°C in a 5% CO2 incubator.
|
[] |
ChemBL_V1
|
The absorbance at 570 nm was measured using a Multiskan SkyHigh Plate Reader by subtracting the absorbance at 630 nm from the background after the purple formazan crystals completely dissolved.
|
[] |
ChemBL_V1
|
Using GraphPad Prism Version 8.0, a log graph of the log test item concentration vs. cell survival percentage was plotted and the half-maximal inhibitory concentration (IC50) values were calculated.
|
[] |
ChemBL_V1
|
.(1)Percent cell survival %=Absorbance of TestAbsorbance of ControlX 100.
|
[] |
ChemBL_V1
|
All statistical analyses were carried out using the SPSS® statistical software package for Windows®, version 15.0 of SPSS Inc. (Chicago, IL, USA).
|
[] |
ChemBL_V1
|
The results are presented as means ± SD, and p ≤ 0.05 was used to determine whether treatment differences were significant.
|
[] |
ChemBL_V1
|
The MTT assay is widely used to assess cell viability, cell proliferation, cytotoxicity testing, and drug screening .
|
[] |
ChemBL_V1
|
In drug screening, the MTT assay is used to test the cytotoxicity of various compounds, such as drugs, natural products, or experimental molecules, on cell lines or primary cells .
|
[] |
ChemBL_V1
|
It measures the reduction of the MTT reagent to formazan, a purple-colored product indicating the metabolic activity of viable cells and the intensity of which is directly proportional to the number of viable cells in the sample .
|
[] |
ChemBL_V1
|
Thus, it helps to identify compounds that inhibit cell growth or induce cell death.
|
[] |
ChemBL_V1
|
In this study, the comparative evaluation for anticancer potential of W. somnifera leaf, stem, and root methanolic extracts was carried out to determine their cytotoxic effect on the selected cancer cell line HepG2 and normal cell line L929 viability.
|
[
{
"end": 208,
"label": "CellLine",
"start": 192,
"text": "cancer cell line"
},
{
"end": 208,
"label": "CellLine",
"start": 192,
"text": "cancer cell line"
},
{
"end": 214,
"label": "CellLine",
"start": 209,
"text": "HepG2"
},
{
"end": 240,
"label": "CellLine",
"start": 236,
"text": "L929"
}
] |
ChemBL_V1
|
HepG2 and L929 cells were treated with W. somnifera leaf, stem, and root methanolic extracts of 20, 50, 100, 200, 500, 1000, 1500, and 2000 μg/mL for 24 h. Figures 2 and 3 show the in vitro morphological profiles of HepG2 and L929 cells, respectively, under an inverted microscope.
|
[
{
"end": 5,
"label": "CellLine",
"start": 0,
"text": "HepG2"
},
{
"end": 14,
"label": "CellLine",
"start": 10,
"text": "L929"
},
{
"end": 221,
"label": "CellLine",
"start": 216,
"text": "HepG2"
},
{
"end": 230,
"label": "CellLine",
"start": 226,
"text": "L929"
}
] |
ChemBL_V1
|
The treatments were compared to controls (a negative sample without a test sample and a positive sample with the standard drug doxorubicin).
|
[] |
ChemBL_V1
|
Cell morphology revealed significant morphological changes in HepG2 and L929 cells.
|
[
{
"end": 67,
"label": "CellLine",
"start": 62,
"text": "HepG2"
},
{
"end": 76,
"label": "CellLine",
"start": 72,
"text": "L929"
}
] |
ChemBL_V1
|
The HepG2 cells showed adhesion to the well wall with W. somnifera methanolic extracts of root at 20 μg/mL, leaf at 20 μg/mL, and stem at 50 μg/mL. On the other hand, L929 cells showed adherence to the well plate wall with W. somnifera methanolic extracts of root at 100 μg/m L, leaf at 100 μg/mL, and stem at 200 μg/mL. However, at higher concentrations, the HepG2 and L929 cells appeared to be in a state of splitting, detaching from the substrate, and increasing the number of suspended cells.
|
[
{
"end": 9,
"label": "CellLine",
"start": 4,
"text": "HepG2"
},
{
"end": 171,
"label": "CellLine",
"start": 167,
"text": "L929"
},
{
"end": 365,
"label": "CellLine",
"start": 360,
"text": "HepG2"
},
{
"end": 374,
"label": "CellLine",
"start": 370,
"text": "L929"
}
] |
ChemBL_V1
|
The viability of HepG2 and L929 cells was evaluated using the MTT assay.
|
[
{
"end": 22,
"label": "CellLine",
"start": 17,
"text": "HepG2"
},
{
"end": 31,
"label": "CellLine",
"start": 27,
"text": "L929"
}
] |
ChemBL_V1
|
A considerable reduction in cell viability was observed in HepG2 (Tables 1–3) in a concentration-dependent manner.
|
[
{
"end": 64,
"label": "CellLine",
"start": 59,
"text": "HepG2"
}
] |
ChemBL_V1
|
At the highest 2000 μg/mL concentration of W. somnifera leaf, stem, and root methanolic extracts, MTT assay results indicated that 100% viability in control HepG2 cells decreased to 10.19 ± 0.015%, 9.93 ± 0.051%, and 9.82%, respectively (Tables 1–3).
|
[
{
"end": 162,
"label": "CellLine",
"start": 157,
"text": "HepG2"
}
] |
ChemBL_V1
|
Moreover, the viability of HepG2 cells at the lowest concentration of 20 μg/mL W. somnifera methanolic leaf, stem, and root extracts was observed to be 81.94 ± 0.046%, 86.09 ± 0.020%, and 95.99 ± 0.025%, respectively.
|
[
{
"end": 32,
"label": "CellLine",
"start": 27,
"text": "HepG2"
}
] |
ChemBL_V1
|
These values were much lower than the viability of the control cells (untreated cells plus media), which ranged between 99 and 100%.
|
[] |
ChemBL_V1
|
Similarly, at the highest 2000 μg/mL concentration of W. somnifera leaf, stem, and root methanolic extracts, the viability of L929 cells decreased from 100% in controls to 26.53 ± 0.045%, 25.94 ± 0.026%, and 26.87 ± 0.058%, respectively (Tables 4–6).
|
[
{
"end": 130,
"label": "CellLine",
"start": 126,
"text": "L929"
}
] |
ChemBL_V1
|
In contrast, at the minimal concentration of 20 μg/mL W. somnifera leaf, stem, and root methanolic extracts, the viability of L929 cells was 72.37 ± 0.030%, 71.97 ± 0.052%, and 77.96 ± 0.030%, respectively.
|
[
{
"end": 130,
"label": "CellLine",
"start": 126,
"text": "L929"
}
] |
ChemBL_V1
|
These values were greater than the values of HepG2 cell viability.
|
[
{
"end": 50,
"label": "CellLine",
"start": 45,
"text": "HepG2"
}
] |
ChemBL_V1
|
However, these values were much lower than the 99–100% viability of the control cells (untreated cells + media).
|
[] |
ChemBL_V1
|
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