@article {1281, title = {Screening of in vitro Anticancer Activity of Various Human Cancer Cell Lines and Induced Apoptosis and Cell Arrest by Ethanolic Bulb Extract of Pancratium zeylanicum}, journal = {Pharmacognosy Research}, volume = {16}, year = {2024}, month = {October 2024}, pages = {718-726}, type = {Original Article}, chapter = {1}, abstract = {

Aim: World widely the human society suffers from various types of cancer due to many causes. Thus, finding out a good anticancer agent is a need of the hour. Medicinal herbs are always considered at pole position whenever we discuss anticancer properties. Numerous herbal plants have been assessed and are subjected to phytochemical analysis to learn more about their anticancer effects. Materials and Methods: To assess the anti-cancerous activity of PZBE extract in MTT assay, cell cycle analysis, and apoptosis studies have been performed using different cancer cell lines. Results: This study aims to assess the impact of ethanolic extract of Pancratium zeylanicum Bulbs (PZBE) against cancerous cells in vitro conditions. MTT assay was used to compare the PZBE extract with Annexin V as a positive control compound against four human cancer cell lines, including Human cervical cancer (HeLa), colon cancer (HT-29), lung cancer (A549) and breast cancer (MCF-7) cell lines with the IC50 values of 209.28 μg/mL for HeLa, 133.09 μg/mL for A-549, 264.46 μg/mL for MCF-7 and 264.75 μg/mL for HT-29 cell lines. Increasing of the populations observed in G2-M and G0-G1 cells by 42\% and 44.78\%, respectively when compared to the control. The graph demonstrates that PZBE extracts significantly induced G0-G1 arrest in A-549 cells. Furthermore, the PZBE extract effectively induced apoptosis when compared to untreated cells, A-549 cells treated with IC50 concentration, and annexin-V double-staining assay demonstrated an increase in the percentage of early and late apoptotic cells. Conclusion: The ethanol extract of Pancratium zeylanicum bulbs demonstrated potent cytotoxic activity against the cancer cell lines HeLa, A-549, MCF-7, and HT-29. The current study{\textquoteright}s findings may be helpful in the development of plant-based anticancer drugs.

}, keywords = {A-549, Annexin-V, Anticancer activity, Apoptosis, Ethanol extract, HeLa, HT-29, MCF-7}, doi = {10.5530/pres.16.4.83}, author = {Elumalai Shanthi Priya and Devaraj Sathiya Sheela and Devaraj Sheeba Gnanadeebam and Balrajmani Davidson Raja and Periannan Viswanathan} } @article {1185, title = {Acacia nilotica Pod Extract has an Anti-cancer Effect on the U937 Cell Line}, journal = {Pharmacognosy Research}, volume = {15}, year = {2023}, month = {October 2023}, pages = {771-775}, type = {Original Article}, chapter = {771}, abstract = {

Background: Acacia nilotica (A. nilotica), a tree that thrives in tropical regions, has been reported to have various medicinal uses and various pharmacological properties. Potential anti-cancer effects are one of the medical properties of parts of this tree. Objectives: To explore and evaluate the cytotoxic effect of A. nilotica, we used Acacia nilotica{\textquoteright}s pod extract on U937 cell lines as an acute myeloid leukemia model. Materials and Methods: Microscopic analysis, flow cytometry, and WST-1 tests were performed on the U937 cell line after 72 hr of treatment with different concentrations of ethanol pod extract from Acacia nilotica (A. nilotica) to test for DNA cell cycle, cell viability and apoptosis, and to determine the IC50 of the drug. Results: Treatment with A. nilotica pod extract significantly induced cell death in U937-treated cells compared with untreated control cells. Microscopic analysis revealed morphological changes in dead and fragmented cells associated with a reduction in cell number. A reduction in cell viability with an increase of cells in the late apoptosis stage by flow cytometry indicated the cytotoxic effect of A. nilotica and accumulation of cells in the sub-G1 phase of the DNA cell cycle analysis. The IC50 was identified by the WST-1 assay. Conclusion: The findings of the present study suggest that A. nilotica may be employed as a natural anti-cancer agent in research on acute myeloid leukemia in humans.

}, keywords = {Apoptosis, cytotoxicity, Flow cytometry, Natural drugs}, doi = {10.5530/pres.15.4.081}, author = {Halla Falih Bakheit and Sebastien Taurin and Elwaleed Mohamed Elamin and Moiz Bakhiet} } @article {1099, title = {Emodin Exhibits Strong Cytotoxic Effect in Cervical Cancer Cells by Activating Intrinsic Pathway of Apoptosis}, journal = {Pharmacognosy Research}, volume = {14}, year = {2022}, month = {October 2022}, pages = {499-503}, type = {Original Article}, chapter = {499}, abstract = {

Background: Cervical cancer is widely acknowledged as the main cause of mortality and a major obstacle to increasing life expectancy, after breast cancer, in women. Natural products have been proven to be a promising source for the development of potential anticancer drugs. Emodin (1,3,8-trihydroxy-6-methyl-9,10-anthracenedione) is shown to exert wide range of biological effects, including antibacterial, hepatoprotective, anti-inflammatory and anticancer. Aim: The major aim of this study was to evaluate the efficacy of emodin against the progression of human cervical cancer. Materials and Methods: The cytotoxic effect of emodin on cervical cancer HeLa cells was assessed by cell viability assay and phase contrast microscopy. Moreover, DAPI staining was performed to analyze nuclear condensation. H2DCFDA staining was done to evaluate ROS generation. Activation of caspases was determined to ascertain mitochondria-mediated apoptosis in cervical cancer cells. Results: The results revealed that emodin strongly inhibited the HeLa cell growth and proliferation; which was validated by cell viability assay and phase contrast microscopy. Moreover, DAPI staining authenticated nuclear condensation and fragmentation in HeLa cells indicating initiation of apoptosis. We also observed significant ROS generation and caspases activation in emodin-treated cervical cancer cells. Conclusion: Our results suggested strong cytotoxic potential of emodin against cervical cancer cells.

}, keywords = {Apoptosis, Cervical Cancer, DAPI, Emodin, HeLa, ROS}, doi = {10.5530/pres.14.4.72}, author = {Amir Saeed and Ahmed Alharbi} } @article {1061, title = {KRAS G12C as a Target of Naringenin for Inducing Cell Death in NCI-H23 Cells}, journal = {Pharmacognosy Research}, volume = {14}, year = {2022}, month = {July 2022}, pages = {256-262}, type = {Original Article}, chapter = {256}, abstract = {

Background: Naringenin (NGN) is a commonly available flavonoid in the citrus fruits. We have previously shown that NGN is cytotoxic to the non-small cell lung cancer (NSCLC) cell line NCI-H23 (H23). Objectives: To check whether NGN could bind to the Kirsten rat sarcoma (KRAS) G12C mutant and cause its inhibition to promote apoptosis in H23 cells. Materials and Methods: NGN was docked with mutant KRAS protein followed by molecular dynamics simulation. HDOCK was used to analyse the influence of NGN on the KRAS and PI3K proteinprotein docking. We checked the ultramorphological structure of the cells. A 2D-QSAR study was carried out to predict the activity of NGN. Results: We observed that NGN bound stably to the mutant KRAS. NGN showed steady RMSF and RMSD values, good structural stability, and favourable MM/PBSA values. NGN interfered in the binding of KRAS and PI3K. NGN treated cells showed hallmarks of apoptotic cell death. The predictive pIC50 value was found to be 7.39 for NGN against KRAS. NGN cleared all the drug filters. Conclusion: We conclude that NGN could bind to the mutant KRAS potentially inhibiting KRAS. That affects the PI3K/Akt pathway activation leading to apoptosis in the NCI-H23 cells.

}, keywords = {Apoptosis, Docking, KRAS, naringenin, NSCLC, QSAR}, doi = {10.5530/pres.14.3.37}, author = {Abani Kumar Patar and Lakhon Kma and Jitul Barman and Shekhar Ghosh and Taranga Jyoti Baruah} } @article {291, title = {The Hydroalcoholic Extract of Matricaria chamomilla Suppresses Migration and Invasion of Human Breast Cancer MDA-MB-468 and MCF-7 Cell Lines}, journal = {Pharmacognosy Research}, volume = {9}, year = {2021}, month = {February 2017}, pages = {87-95}, type = {Original Article}, chapter = {87}, abstract = {

Background: Matricaria chamomilla is an aromatic plant with antioxidant, anticancer, and anti-inflammatory properties. However, the inhibitory role of M. chamomilla on migration and invasion of human breast cancer cells remains unclear. gThis study investigated the methods to evaluate these anticancer mechanisms of M. chamomilla on human breast cancer MCF-7 and MDA-MB-468 cell lines. Materials and Methods: The cells were treated with hydroalcoholic extract of M. chamomilla at different concentrations (50{\textendash}1300 μg/mL) for 24, 48, and 72 h in a culture medium containing 10\% fetal bovine serum. This study quantified the 50\% growth inhibition concentrations (IC50) by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay; apoptosis and necrosis through Hoechst 33342/propidium iodide staining; cell proliferation and clone formation by clonogenic assay as well as cellular migration, invasion, and attachment. After 24, 48, and 72 h of treatment, the IC50 levels were 992 {\textpm} 2.3 μg/mL, 893 {\textpm} 5.4 μg/ mL, and 785 {\textpm} 4.8 μg/mL against MDA-MB-468, respectively, and 1288 {\textpm} 5.6 μg/mL, 926 {\textpm} 2.5 μg/mL, and 921 {\textpm} 3.5 μg/mL, against MCF-7, respectively. Furthermore, increasing the extract concentrations induced cellular apoptosis and necrosis and decreased cellular invasion or migration through 8 μm pores, colonization and attachment in a dose-dependent manner. Results: It indicated time- and dose-dependent anti-invasive and antimigrative or proliferative and antitoxic effects of hydroalcoholic extract of aerial parts of chamomile on breast cancer cells. Conclusion: This study demonstrated an effective plant in preventing or treating breast cancer.

}, keywords = {Apoptosis, Attachment, Breast cancer, Invasive, Migration}, doi = {10.4103/0974-8490.199778}, author = {Mohsen Nikseresht, and Ali Mohammad Kamali and Hamid Reza Rahimi and Hamdollah Delaviz and Mehdi Akbartabar Toori and Iraj Ragerdi Kashani and Reza Mahmoudi} } @article {89, title = {Induction of Triple-negative Breast Cancer Cells to Immunogenic Cell Death and Increase Cross-Presentation by Streptomyces sanyensis}, journal = {Pharmacognosy Research}, volume = {13}, year = {2021}, month = {June 2021}, pages = {165-172}, type = {Original Article}, chapter = {165}, abstract = {

Background: Biomass extract of Streptomyces sanyensis (BE-SS), which is a marine microorganism, has antitumor activity and has white-to-gray aerial mycelium and long chains of long spores in aerial mycelium. Objectives: The anti-cancer effect of BE-SS on triple-negative breast cancer (TNBC) was confirmed and cell death by BE-SS was confirmed to have immunogenicity and whether it could be used for immuno-cancer therapy. Materials and Methods: Human and mouse TNBC cells (MDA-MB-231, HS578T, 4T1-hemagglutinin (HA) and TUBO-P2J-HA cells) were treated with BE-SS at different concentrations for 72 hr and their cytotoxicity was detected using the sulforhodamine B-based (SRB) method. Flow cytometry was used to determine the type of cell death by Annexin V/7-AAD staining and to measure surface exposure to damage-related molecular pattern (DAMP) molecules including calreticulin (CRT), heat shock protein (HSP) 70/90. Carboxyfluorescein succinimidyl ester (CFSE) dilution assay was used to evaluate the immunogenicity of dead cells induced by BE-SS. Results: BE-SS reduced the viability of human (MDA-MB-231 and HS578T-cells) and mouse (4T1-HA and TUBO-P2J-HA cells) breast cancer cells. At 72 h, the IC50 of human and mouse breast cancer cells was 0.02-0.6 μg/ml and 0.005-0.37 μg/ml, respectively. BE-SStreated breast cancer cells were positively stained with Annexin V. Surface exposure to DAMP molecules increased in a dose-and time-dependent manner. CFSE dilution analysis showed that dendritic cells (DCs) fed with BE-SS treated breast cancer cells successfully stimulated tumor-specific T-cell proliferation without inhibiting DC function and T-cell proliferation. Conclusion: BE-SS can induce immunogenicity and apoptosis in breast cancer cells and may be a good adjuvant for TNBC immunotherapy.

}, keywords = {Apoptosis, Cross-presentation, DAMPs, Immunogenic cell death, Streptomyces sanyensis, TNBC}, doi = {10.5530/pres.13.3.9}, author = {Xingguo Quan and Ji-Young Lee and Jin Hee Park and Md. Masudul Haque and Hee Yeon Kim and Ilhwan Kim and Anbok Lee and Il-Whan Choi and SaeGwang Park} } @article {64, title = {Anticancer Activity of Ethyl Acetate Extract from Lethariella cladonioides in vitro and in vivo}, journal = {Pharmacognosy Research}, volume = {12}, year = {2020}, month = {February 2020}, pages = {40-46}, type = {Original Article}, chapter = {40}, abstract = {

Background: Lethariella cladonioides, also known as Red Snow Tea and Golden Silk Tea, mainly grows in Tibet, Sichuan and Yunnan of China. Its medicinal value has been gradually discovered. Objectives: This study mainly evaluated the anti-cancer effect of AMH-L extracted from L. cladonioides in vitro and in vivo and to explore its anticancer mechanism. Materials and Methods: MTT assay was used to determine the cytotoxicity of AMH-L to human cancer cells A-549 and HCT-116 in vitro. Sarcoma S-180 and human lung cancer NCL-H460 cells were implanted into Kunming mice and Balb/C nude mice, respectively, anti-cancer activity in vivo was assessed by recording body weight, organ coefficient and tumor inhibition rate. Flow cytometry and western blotting were used to preliminarily explore the anticancer mechanism. Results: AMH-L, the active component of Red Snow Tea, had strong anticancer activity against A-549 and HCT-116, and IC50 was 26.17 {\textpm} 3.54 and 16.21 {\textpm} 0.83. When the dosage was 50, 100 and 150 mg/ kg, the inhibition rates of S-180 in Kunming mice were 36.04\%, 43.14\%, and 50.45\% in turn, respectively. At the end of the experiment, the dose was 150, and the relative proliferation inhibition rate of Balb/C nude mice NCI-H460 tumor was 50.36\%. Cell cycle and apoptotic results showed that AMH-L could block A-549 at G2/M phase and promote apoptosis of HCT-116 by inducing poly (ADP-ribose) polymerase lysis. Conclusion: L. cladonioides extract AMH-L exhibits strong anticancer activity in vivo and in vitro and can be used as a potential anticancer drug for further study.

}, keywords = {Anticancer drug, Apoptosis, Cell cycle, Cleaved poly (ADP-ribose) polymerase, Lichen}, doi = {10.4103/pr.pr_70_19}, author = {Wen-Jing Lian and Xiao-Qiong He and Qian Yao and Zhong-Yu Song and Jing Wang} } @article {211, title = {Antihypertensive Effect of Methanol Leaf Extract of Azadirachta indica is Mediated through Suppression of Renal Caspase 3 Expressions on Nω-Nitro-L-Arginine Methyl Ester Induced Hypertension}, journal = {Pharmacognosy Research}, volume = {12}, year = {2020}, month = {January 2021}, pages = {460-465}, type = {Original Article}, chapter = {460}, abstract = {

Background: Azadirachta indica (AI) Adr Juss (Meliaceae), known as neem, has been used traditionally for the treatment of various disease conditions including obesity and hypertension. Objective: The antihypertensive effect and mechanism of action of modulatory effect of AI were investigated after the induction of hypertension using Nω-nitro-L-arginine methyl ester (L-NAME). Materials and Methods: Five groups of ten rats divided as follows: Control; L-NAME (40 mg/kg); L-NAME + 100 mg/kg AI; L-NAME and 200 mg/kg AI; and L-NAME and Enalapril (25 mg/kg) were used. Results: following the application of L-NAME, hypertension (elevated systolic, diastolic, mean arterial blood pressures) and increased levels of oxidative stress markers were observed in rats. Immunohistochemistry showed increased caspase-3 expressions in hypertensive rats compared to normotensive rats. Conversely, AI treatment resulted in restoration of physiological antioxidant status and normotension, comparable to the standard antihypertensive agent enalapril. Conclusion: AI leaf is a good candidate for the management of high blood pressure.

}, keywords = {Antioxidant, Apoptosis, Azadirachta indica, Hypertension, Oxidative stress, Phytotherapy}, doi = {10.4103/pr.pr_10_20}, author = {Temidayo Olutayo Omobowale and Ademola Adetokunbo Oyagbemi and Olumuyiwa Abiola Adejumobi and Fisayo Ugbor and Ebunoluwa Racheal Asenuga and Temitayo Olabisi Ajibade and Jeremiah Moyinoluwa Afolabi and Blessing Seun Ogunpolu and Olufunke Olubunmi Falayi and Idayat Titilayo Gbadamos and Olufunke Eunice Ola-Davies and Adebowale Bernard Saba and Anofi Ashafa and Momoh Audu Yakubu and Adeolu Alex Adedapo and Oluwafemi Omoniyi Oguntibeju} } @article {284, title = {Cytotoxicity and Apoptosis Induction of Sea Cucumber Holothuria atra Extracts}, journal = {Pharmacognosy Research}, volume = {11}, year = {2019}, month = {February,2019}, pages = {41-46}, type = {Original Article}, chapter = {41}, abstract = {

Context: Cancer is one of the major causes of death. Sea cucumbers are marine invertebrate that is widely used and utilized in traditional medicine. One of the most studied of sea cucumbers efficacy is its anticancer property. The anticancer property of sea cucumbers is much related to the content of an active compound called saponin. Objective: The objective of this research is to determine the cytotoxicity and apoptosis induction of sea cucumber Holothuria atra ethanol extract and to study its anticancer active compound. Materials and Methods: Sea cucumber of H. atra was taken from Halmahera waters, North Maluku Indonesia. The extraction was conducted by maceration method using ethanol 96\% then continued with liquid-liquid partition and separation using C18 column. The investigation of active compound content was conducted by phytochemical and spectroscopic analysis using liquid chromatography-ion trap-time of a flight-mass spectrophotometer (LC-IT-ToF-MS). The cytotoxicity test was performed by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide method against human breast ductal carcinoma cell (T47D), human breast adenocarcinoma (MCF7), human colorectal adenocarcinoma cell (WiDr), and human cervix adenocarcinoma cell lines (HeLa). The apoptotic induction test was performed using flow cytometry using Fluorescein isothiocyanate (FITC) Annexin-V-Apoptosis and caspase-3 detection using FITC Active Caspase-3 kit. Results: Phytochemical test showed that ethanol extract of H. atra contained alkaloids, flavonoids, steroids-triterpenoids, phenols, and saponins. The extract showed cytotoxic activity against the 4 cell lines with Inhibition concentration 50 values ranging from 9.6 to 14.3 μg/ml. Flow cytometry analysis showed that the T47D cell population underwent apoptosis after treated with ethanol extract. The extract also activated caspase-3 on the T47D cells. The results of LC-IT-ToF-MS analysis showed that the active fraction from C18 column separation contained saponin and identified as Cucumechinol and Philinopgenin B. Conclusion: The results of this study indicated that H. atra active extract has good cytotoxicity and has potential to be developed as an anticancer agent.

}, keywords = {Apoptosis, caspase-3, cytotoxicity, Holothuria atra, Saponin, Sea cucumber.}, doi = {10.4103/pr.pr_3_18}, author = {Muhammad Nursid and Endar Marraskuranto and Ekowati Chasanah} } @article {140, title = {Cytotoxicity and Apoptosis Induction of Sea Cucumber Holothuria atra Extracts}, journal = {Pharmacognosy Research}, volume = {11}, year = {2019}, month = {February 2019}, pages = {41-46}, type = {Original Article}, chapter = {41}, abstract = {

Context: Cancer is one of the major causes of death. Sea cucumbers are marine invertebrate that is widely used and utilized in traditional medicine. One of the most studied of sea cucumbers efficacy is its anticancer property. The anticancer property of sea cucumbers is much related to the content of an active compound called saponin. Objective: The objective of this research is to determine the cytotoxicity and apoptosis induction of sea cucumber Holothuria atra ethanol extract and to study its anticancer active compound. Materials and Methods: Sea cucumber of H. atra was taken from Halmahera waters, North Maluku Indonesia. The extraction was conducted by maceration method using ethanol 96\% then continued with liquid-liquid partition and separation using C18 column. The investigation of active compound content was conducted by phytochemical and spectroscopic analysis using liquid chromatography-ion trap-time of a flight-mass spectrophotometer (LC-IT-ToF-MS). The cytotoxicity test was performed by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide method against human breast ductal carcinoma cell (T47D), human breast adenocarcinoma (MCF7), human colorectal adenocarcinoma cell (WiDr), and human cervix adenocarcinoma cell lines (HeLa). The apoptotic induction test was performed using flow cytometry using Fluorescein isothiocyanate (FITC) Annexin-V-Apoptosis and caspase-3 detection using FITC Active Caspase-3 kit. Results: Phytochemical test showed that ethanol extract of H. atra contained alkaloids, flavonoids, steroids-triterpenoids, phenols, and saponins. The extract showed cytotoxic activity against the 4 cell lines with Inhibition concentration 50 values ranging from 9.6 to 14.3 μg/ml. Flow cytometry analysis showed that the T47D cell population underwent apoptosis after treated with ethanol extract. The extract also activated caspase-3 on the T47D cells. The results of LC-IT-ToF-MS analysis showed that the active fraction from C18 column separation contained saponin and identified as Cucumechinol and Philinopgenin B. Conclusions: The results of this study indicated that H. atra active extract has good cytotoxicity and has potential to be developed as an anticancer agent.

}, keywords = {Apoptosis, caspase-3, cytotoxicity, Holothuria atra, Saponin, Sea cucumber}, doi = {10.4103/pr.pr_3_18}, author = {Muhammad Nursid and Endar Marraskuranto and Ekowati Chasanah} } @article {276, title = {Therapeutic Implications of Ajwa Dates (Phoenix dactylifera) in the Inhibition of Liver Tissue Alterations through the Modulation of Vascular Endothelial Growth Factor and Phosphatase, and Tensin Homolog Gene}, journal = {Pharmacognosy Research}, volume = {10}, year = {2018}, month = {April 2018}, pages = {151-155}, type = {Original Article}, chapter = {151}, abstract = {

Background: Dates are a plant species in the palm family, Arecaceae and is used as staple food in the Middle East for thousands of years. Aim: The current study was undertaken to evaluate the protective role of ajwa dates extract (ADE) on carbon tetrachloride (CCl4)-induced hepatotoxicity. Materials and Methods: The study was carried out on mice mode through different groups as control group without treatment of CCl4, ADE and CCl4-treated group, and CCl4-treated group only. Results: This finding demonstrated that histological alterations including degeneration, congestion and infiltration of lymphocytes were seen in the liver tissue in CCl4-treated groups. However, ADE-treated groups showed protection to attenuate CCl4-induced liver toxicity through maintenance of architecture of hepatocytes as evident of congestion, necrosis, and degeneration was not noticed. Moreover, a few number of infiltrations of lymphocytes were noticed in ADE-treated groups. However, expression of phosphatase and tensin homolog (PTEN) and vascular endothelial growth factor (VEGF) protein was evaluated in all groups, and it was observed that PTEN was highly expressed in control group and ADE-treated group. Whereas, PTEN expression was also observed in CCl4-treated group, but out of eight cases, one case showed less expression of PTEN protein. The difference in expression pattern of PTEN protein in CCl4-treated group and ajwa-treated group was statically insignificant (P \> 0.05). VEGF was not expressed in control and ajwa-treated group. While expression of VEGF protein was observed in CCl4-treated group, and it was noticed that two out of eight cases showed expression of VEGF protein. Conclusion: The findings supported the idea that ADE might reduce liver tissue alterations or maintenance of architecture of hepatocytes. Ajwa dates-treated group showed decreased in the VEGF expression and such angiogenesis process involve in migration and differentiation of endothelial cells as well as prevent the loss of PTEN protein expression.

}, keywords = {Ajwa dates, Apoptosis, Carbon tetrachloride, Hepatoprotective activity, Phosphatase and tensin homolog, Vascular endothelial growth}, doi = {10.4103/pr.pr_119_17}, author = {Arshad H. Rahmani and Amjad A. Khan and Ali Y. Babiker and Masood A. Khan and Mohammad A. Alzohairy} } @article {456, title = {Annonacin Exerts Antitumor Activity through Induction of Apoptosis and Extracellular Signal-regulated Kinase Inhibition}, journal = {Pharmacognosy Research}, volume = {9}, year = {2017}, month = {November 2017}, pages = {378-383}, type = {Original Article}, chapter = {378}, abstract = {

Background: Endometrial cancer (EC) is the most common gynecologic malignancy in developed countries. Annonacin, a natural pure compound extracted from the seeds of Annona muricata, is a potential alternative therapeutic agent to treat EC. Objective: To study the antitumor activity of annonacin and its mechanism of action in EC cells (ECCs). Materials and Methods: Viability of ECCs treated with annonacin for 72 h was determined using methyl thiazolyl tetrazolium assay. The induction of cell cycle arrest and apoptotic cell death was evaluated using propidium iodide and annexin V-PE/7-AAD assay, respectively. DNA strand breaks were visualized using transferase dUTP nick end labeling assay, and the effects of annonacin on survival signaling were determined using western blotting. Results: Annonacin exhibited antiproliferative effects on EC cell lines (ECC-1 and HEC-1A) and primary cells (EC6-ept and EC14-ept) with EC50 values ranging from 4.62 to 4.92 {\textmu}g/ml. EC cells were shown arrested at G2/M phase after treated with 4 {\textmu}g/ml of annonacin for 72 h. This led to a significant increase in apoptotic cell death (65.7\%) in these cells when compared to vehicle-treated cells (P \< 0.005). We further showed that annonacin-mediated apoptotic cell death was associated with an increase in caspase-3 cleavage and DNA fragmentation. Cell apoptosis was accompanied with downregulation of extracellular signal-regulated kinase survival protein expression and induction of G2/M cell cycle arrest. Conclusion: Annonacin may be a potential novel therapeutic agent for EC patients.

}, keywords = {Annona muricata, Apoptosis, caspase-3, Cell cycle arrest, Extracellular signal-regulated kinase, Uterine cancer}, doi = {10.4103/pr.pr_19_17}, author = {Chee Voon Yap and Kavita S Subramaniam and Sik Wey Khor and Ivy Chung} } @article {503, title = {Hepatoprotective and Cytotoxic Activities of Abietic Acid from Isodon wightii (Bentham) H. Hara}, journal = {Pharmacognosy Research}, volume = {8}, year = {2016}, month = {May 2016}, pages = {206-208}, type = {Short Communication}, chapter = {206}, abstract = {

Isodon (Lamiaceae) is a known source of bioactive terpenoids. Diterpenoids isolated from Isodon wightii (Bentham) H. Hara showed antibacterial, antiacetylcholinesterase, antioxidant, anticancer, and anticarcinogenic activities, etc., Hepatoprotective activity of ABA against lipopolysaccharide (LPS) induced liver injury in BALB/c mice was studied. Cytotoxic activity of ABA was tested against cervical cancer cells (HeLa) using MTT assay followed by propidium iodide (PI) staining to identify apoptosis. Histopathological analysis revealed that 1.5 μg/mL LPS induced liver damage was attenuated by ABA in a dose dependent manner. ABA showed cytotoxicity with IC50 value of 176.28 {\textpm} 0.02 μg/mL and PI staining of treated cells showed apoptosis. This study proves that ABA would be a promising natural compound for herbal drug preparation.

}, keywords = {Abietic acid, Apoptosis, BALB/c mice, HeLa, lipopolysaccharide}, doi = {10.4103/0974-8490.182920}, author = {Madhusudhanan Gogul Ramnath and Ramaraj Thirugnanasampandan and Sadasivam Mathusalini and Palathurai Subramanium Mohan} } @article {511, title = {Kayeassamin A Isolated from the Flower of Mammea siamensis Triggers Apoptosis by Activating Caspase-3/-8 in HL-60 Human Leukemia Cells}, journal = {Pharmacognosy Research}, volume = {8}, year = {2016}, month = {August 2016}, pages = {244-248}, type = {Original Article}, chapter = {244}, abstract = {

Background: Mammea siamensis (Miq.) T. Anders. is used as a medicinal plant in Thailand and has several traditional therapeutic properties. In a previous study, we isolated eight compounds from the flower of M. siamensis and demonstrated that kayeassamin A (KA) exhibited potent antiproliferative activity against human leukemia and stomach cancer cell lines. Objective: In this study, we investigated the effect of KA on cell viability and apoptotic mechanisms in HL-60 human leukemia cells. Materials and Methods: Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Nuclear morphology and DNA fragmentation were observed using Hoechst 33258 staining and agarose gel electrophoresis, respectively. The sub-G1 phase of cells was analyzed by flow cytometry after the cellular DNA had been stained with propidium iodide. The protein levels of poly (ADP-ribose) polymerase (PARP) and caspases were determined by Western blotting. Results: KA exhibited a significant cytotoxic effect in a dose- and time-dependent manner, and induced chromatin condensation, DNA fragmentation, and sub-G1 phase DNA content, known as molecular events associated with the induction of apoptosis. In addition, KA strongly induced the activation of PARP and caspase-3 and -8, with weak caspase-9 activation. Furthermore, KA-induced DNA fragmentation was abolished by pretreatment with z-VAD-FMK (a broad caspase inhibitor), z-DEVD-FMK (a caspase-3 inhibitor), and z-IETD-FMK (a caspase-8 inhibitor), but not by z-LEHD-FMK (a caspase-9 inhibitor) pretreatment. Conclusion: These results indicate that KA triggers apoptotic cell death by activation of caspase-3 and -8 in HL-60 cells.

}, keywords = {Apoptosis, Caspase, Kayeassamin A, leukemia, Mammea siamensis}, doi = {10.4103/0974-8490.188884}, author = {Takuhiro Uto and Nguyen Huu Tung and Pinjutha Thongjankaew and Sorasak Lhieochaiphant and Yukihiro Shoyama} } @article {574, title = {Induction of rat hepatic mitochondrial membrane permeability transition pore opening by leaf extract of Olax subscorpioidea}, journal = {Pharmacognosy Research}, volume = {7}, year = {2015}, month = {June 2015}, pages = {s63-s68}, type = {Original Article}, chapter = {63}, abstract = {

Background: The induction of the mitochondrial membrane permeability transition (MMPT) pore has been implicated in the cascade of events involved in apoptosis (programmed cell death). Olax subscorpioidea is traditionally used for the treatment of several diseases and infection. However, its role on MMPT is not yet established. This study was aimed at evaluating the effects of varying concentrations of the methanol leaf extract of O. subscorpioidea (MEOS) on MMPT pore opening, mitochondrial adenosine triphosphatase (ATPase), and mitochondrial lipid peroxidation. Materials and Methods: Opening of the pore was spectrophotometrically assayed under succinate-energized conditions. Results: In the absence of triggering agent (calcium), MEOS induced MMPT pore opening by 350, 612, 827, 845\% at 36, 60, 86 and 112 μg/ml, respectively. MEOS further induced MMPT pore opening in the presence of a triggering agent by 866, 905, 831, 840, 949\% at 12, 36, 60, 86 and 112 μg/ml, respectively. The extract significantly induced mitochondrial membrane lipid peroxidation in all the concentration used. MEOS also significantly increased mitochondrial ATP hydrolysis by mitochondrial ATPase in all concentration of the extract used. Conclusion: It may be deduced from this results, that MEOS contains certain bioactive components that may find use in pathological conditions that require an enhanced rate of apoptosis.

}, keywords = {Apoptosis, Mitochondrial membrane permeability transition pore, Olax subscorpioidea}, doi = {10.4103/0974-8490.157998}, author = {Oluwatobi Samuel Adegbite and Yetunde Ifeoma Akinsanya and Ayobami Jahdahunsi Kukoyi and Wisdom O Iyanda-Joel and Oluwatoyin O Daniel and Abiodun Humphrey Adebayo} } @article {665, title = {Cellular Responses with Thymoquinone Treatment in Human Breast Cancer cell line MCF-7}, journal = {Pharmacognosy Research}, volume = {5}, year = {2013}, month = {May 2013}, pages = {200-206}, type = {Original Article }, chapter = {200}, abstract = {

Background:Nigella sativa\ or black seed extract has been reported to show various medicinal benefits. Thymoquinone which is an active compound of its seed has been reported to contain anti-cancer properties.\ Objective:\ The study addressed the anti-cancer efficiency of long-term\ in vitro\ treatment with thymoquinone towards human breast cancer cell lines MCF-7.\ Materials and Methods:\ Cell proliferation was determined with CellTiter 96 Aqueous. Non-Radioactive Cell Proliferation Assay Kit. It was followed with trypan blue exclusion test to determine the percentage of viable cells. The study incorporated cell cycle assay to distinguish cell distribution at various cell cycle phases using Cycletest Plus DNA Reagent Kit. The apoptosis detection kit was used to determine the percentage of apoptotic and necrotic cells using flow cytometry.\ Results:\ The 50\% inhibitory concentration (IC\ 50\ ) value determined using the proliferation assay was 25 μM thymoquinone. Late apoptotic cell percentage increased rapidly when treatment duration was increased to 24 h with 25 and 100 μM thymoquinone. Further analysis using cell cycle assay showed thymoquinone inhibition of breast cancer cell proliferation at minimal dose 25 μM and led to S phase arrest significantly at 72 h treatment (P\ = 0.009). It was also noted elevation sub-G\ 1\ peak following treatment with 25 μM thymoquinone for 12 h. Increase in thymoquinone to 50 μM caused G\ 2\ phase arrest at each time-point studied.\ Conclusion:\ In general thymoquinone showed sustained inhibition of breast cancer cell proliferation with long-term treatment. Specificity of phase arrest was determined by thymoquinone dose.

}, keywords = {Apoptosis, cytotoxicity, Long-term Exposure, MCF-7 cell line, Thymoquinone}, doi = {10.4103/0974-8490.112428}, author = {Marjaneh Motaghed and Faisal Muti Al-Hassan and Shahrul Sahul Hamid} }