scholarly journals Myrtus Communis Leaf Extract - A Source of Secondary Metabolites Exhibiting Anticancer and Antimycobacterial Activities

2021 ◽  
Author(s):  
Mushtaq A. Mir ◽  
Serag Eldin Elbehairi ◽  
Lamis Ahmad Memish ◽  
Faris Saif ◽  
Nasreena Bashir ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Secondary Metabolites ◽  
Cancer Cells ◽  
Biofilm Formation ◽  
Diffusion Method ◽  
Dna Flow Cytometry ◽  
Myrtus Communis ◽  
Zone Of Inhibition ◽  
Mycobacterial Infections

Abstract Background: Plant-derived products or extracts are widely used in folk/traditional medicine to treat several, infections, ailments, or disorders. A notable therapeutic herb Myrtus communis, worldwide utilized in the traditional medication for centuries, is an evergreen aromatic and medicinal plant of the Mediterranean region. Materials and methods: The SulphoRhodamine-B assay and DNA flow cytometry were used to investigate the proliferation and subsequent distribution of cells among different phases of the cell cycle. Annexin V-FITC/PI staining coupled with flow cytometry was used to analyze apoptosis and necrosis of the cancer cells. Western blotting detected the expression of pro- and anti-apoptotic proteins. Zone of inhibition and MIC were determined by well diffusion method and microplate alamar blue assay, respectively. Biofilm formation was studied by crystal violet method. For statistical analysis, a two-tailed Student’s t-test of GraphPad Prism 6.0 was used.Results: In this study, the secondary metabolites of M. communis leaves extracted in ethanol showed the highest cytotoxicity and thus the greatest anticancer effects against diverse cancer cell lines of the breast (MCF-7), liver (HepG2), cervix (HeLa), and colon (HCT116) (IC50; ranging from 33 to 83 mg/ml). The cancer cells arrested in the G1 phase of the cell cycle undergo apoptosis. The induction of the latter is mediated by activation of the intrinsic mitochondrial pathway. Furthermore, the extract showed a strong growth inhibitory effect (zone of inhibition; 20.3±1.1 - 26.3±2.5 mm, MIC; 4.88 - 312.5 µg/ml, and MBC; 39.07 - 1250 μg/ml) against several rapid and slow-growing mycobacterial strains that cause tuberculosis and several other mycobacterial infections. The biofilm formation in BSL2 microorganisms, M. smegmatis and S. aureus, is strongly inhibited by the extract. Conclusion: These results suggest that M. communis leaf extract is a potential source of secondary metabolites, which could be developed further as potential anti-cancer and anti-mycobacterial agents to treat diverse types of cancers and mycobacterial infections.

scholarly journals Costunolide Induces Apoptosis via the Reactive Oxygen Species and Protein Kinase B Pathway in Oral Cancer Cells

2021 ◽  
Vol 22 (14) ◽  
pp. 7509
Author(s):  
Hai Huang ◽  
Jun-Koo Yi ◽  
Su-Geun Lim ◽  
Sijun Park ◽  
Haibo Zhang ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Cell Cycle ◽  
Flow Cytometry ◽  
Oral Cancer ◽  
Cancer Cells ◽  
Reactive Oxygen ◽  
Migration And Invasion ◽  
Oxygen Species ◽  
Oral Cancer Cells

Oral cancer (OC) has been attracted research attention in recent years as result of its high morbidity and mortality. Costunolide (CTD) possesses potential anticancer and bioactive abilities that have been confirmed in several types of cancers. However, its effects on oral cancer remain unclear. This study investigated the potential anticancer ability and underlying mechanisms of CTD in OC in vivo and in vitro. Cell viability and anchorage-independent colony formation assays were performed to examine the antigrowth effects of CTD on OC cells; assessments for migration and invasion of OC cells were conducted by transwell; Cell cycle and apoptosis were investigated by flow cytometry and verified by immunoblotting. The results revealed that CTD suppressed the proliferation, migration and invasion of oral cancer cells effectively and induced cell cycle arrest and apoptosis; regarding the mechanism, CTD bound to AKT directly by binding assay and repressed AKT activities through kinase assay, which thereby downregulating the downstream of AKT. Furthermore, CTD remarkably promotes the generation of reactive oxygen species by flow cytometry assay, leading to cell apoptosis. Notably, CTD strongly suppresses cell-derived xenograft OC tumor growth in an in vivo mouse model. In conclusion, our results suggested that costunolide might prevent progression of OC and promise to be a novel AKT inhibitor.

scholarly journals αO-Conotoxin GeXIVA Inhibits the Growth of Breast Cancer Cells via Interaction with α9 Nicotine Acetylcholine Receptors

Marine Drugs ◽  
2020 ◽  
Vol 18 (4) ◽  
pp. 195 ◽  
Author(s):  
Zhihua Sun ◽  
Jiaolin Bao ◽  
Manqi Zhangsun ◽  
Shuai Dong ◽  
Dongting Zhangsun ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Cycle ◽  
Flow Cytometry ◽  
Cell Cycle Arrest ◽  
Cancer Cells ◽  
Cell Line ◽  
Cancer Cell ◽  
Breast Cancer Cell ◽  
Breast Cancer Cells ◽  
Cycle Arrest

The α9-containing nicotinic acetylcholine receptor (nAChR) is increasingly emerging as a new tumor target owing to its high expression specificity in breast cancer. αO-Conotoxin GeXIVA is a potent antagonist of α9α10 nAChR. Nevertheless, the anti-tumor effect of GeXIVA on breast cancer cells remains unclear. Cell Counting Kit-8 assay was used to study the cell viability of breast cancer MDA-MD-157 cells and human normal breast epithelial cells, which were exposed to different doses of GeXIVA. Flow cytometry was adopted to detect the cell cycle arrest and apoptosis of GeXIVA in breast cancer cells. Migration ability was analyzed by wound healing assay. Western blot (WB), quantitative real-time PCR (QRT-PCR) and flow cytometry were used to determine expression of α9-nAChR. Stable MDA-MB-157 breast cancer cell line, with the α9-nAChR subunit knocked out (KO), was established using the CRISPR/Cas9 technique. GeXIVA was able to significantly inhibit the proliferation and promote apoptosis of breast cancer MDA-MB-157 cells. Furthermore, the proliferation of breast cancer MDA-MB-157 cells was inhibited by GeXIVA, which caused cell cycle arrest through downregulating α9-nAChR. GeXIVA could suppress MDA-MB-157 cell migration as well. This demonstrates that GeXIVA induced a downregulation of α9-nAChR expression, and the growth of MDA-MB-157 α9-nAChR KO cell line was inhibited as well, due to α9-nAChR deletion. GeXIVA inhibits the growth of breast cancer cell MDA-MB-157 cells in vitro and may occur in a mechanism abolishing α9-nAChR.

DNA flow cytometry: A new technique for rapid analysis of cell-cycle kinetics in human anagen hairs

1985 ◽  
Vol 277 (4) ◽  
pp. 337-339 ◽  
Author(s):  
E. Deinlein ◽  
H. Schell ◽  
M. Winter ◽  
O. P. Hornstein ◽  
K. Katsuoka
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Rapid Analysis ◽  
New Technique ◽  
Dna Flow Cytometry ◽  
Cell Cycle Kinetics ◽  
A New Technique

Histone Deacetylase Inhibitor Trichostatin A Suppresses Cell Proliferation and Induces Apoptosis by Regulating the PI3K/AKT Signalling Pathway in Gastric Cancer Cells

2020 ◽  
Vol 20 (17) ◽  
pp. 2114-2124
Author(s):  
Xinli An ◽  
Zekun Wei ◽  
Botian Ran ◽  
Hao Tian ◽  
Hongyu Gu ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Cell Cycle ◽  
Flow Cytometry ◽  
Cell Proliferation ◽  
Cancer Cells ◽  
Histone Deacetylase ◽  
Trichostatin A ◽  
Gastric Cancer Cells ◽  
Growth Modulation ◽  
Regulatory Pathways

Background: Gastric cancer, a common malignant tumour worldwide, has a relatively poor prognosis and is a serious threat to human health. Histone Deacetylase Inhibitors (HDACi) are anticancer agents that are known to affect the cell growth of different cancer types. Trichostatin A (TSA) selectively inhibits the class I and II mammalian Histone Deacetylase (HDAC) family enzymes and regulates many cell processes. Still, the underlying mechanisms of HDACs are not fully understood in gastric cancer. Objective: This study aims to investigate the antitumor effect and the mechanism of growth modulation of gastric cancer cells by TSA. Methods: The cell proliferation of gastric cancer cells was measured by MTT and BrdU immunofluorescence assays. Soft agar assay was used to detect the colony formation ability of gastric cancer cells. Flow cytometry was used to examine cell cycle and apoptosis. Western blot was employed to detect protein expression of target factors. Results: TSA inhibits the proliferation of MKN-45 and SGC-7901 cells and leads to significant repression of colony number and size. Flow cytometry assays show TSA induces cell cycle arrest at G1 phase and apoptosis, and TSA effects the expression of related factors in the mitochondrial apoptotic signalling and cell cycle-related regulatory pathways. Furthermore, TSA increased histone H3K27 acetylation and downregulated the expression of PI3K and p-AKT. Conclusion: Downregulating PI3K/AKT pathway activation is involved in TSA-mediated proliferation inhibition of gastric cancer.

Carvacrol Exhibits Chemopreventive Potential against Cervical Cancer Cells via Caspase-Dependent Apoptosis and Abrogation of Cell Cycle Progression

2020 ◽  
Vol 21 ◽  
Author(s):  
Afza Ahmad ◽  
Irfan A. Ansari
Keyword(s):  
Cell Cycle ◽  
Cervical Cancer ◽  
Flow Cytometry ◽  
Cancer Cells ◽  
Cell Cycle Progression ◽  
Natural Compounds ◽  
Cell Cycle Distribution ◽  
Chemotherapeutic Drugs ◽  
Cycle Progression ◽  
Cervical Cancer Cells

Background:: The carcinogenesis of uterine cervix is predominantly initiated with the consistent infection of human papilloma virus (HPV). Owing to the adverse side effects of standard chemotherapeutics in the treatment of recurrent and metastatic cervical cancer, there is a need for better and effective treatment modality. In this lieu of concern, natural compounds have proven their worthwhile potential against treatment of various carcinomas. Carvacrol is a phenolic monoterpenoid and several reports have suggested its different biological properties including antioxidant, anti-inflammatory and anticancer activity. Objective:: The objective of our present study was to investigate the effect of carvacrol on HPV18+ HeLa cervical cancer cells. Methods:: HeLa cervical cancer cells were cultured and subsequently treated with various doses of carvacrol. Cell viability was assessed via MTT assay. DAPI and Hoechst3342 staining was used to qualitatively analyzed the induced apoptosis. Reactive Oxygen Species (ROS) was estimated by DCFDA staining protocol and quantitatively estimated by flow cytometry. The cell cycle distribution and apoptosis (FITC-Annexin V assay) were analyzed by flow cytometry. Results:: The results of the present study have established that carvacrol strongly suppresses proliferation of cervical cancer cells via caspase-dependent apoptosis and abrogation of cell cycle progression. Furthermore, our preliminary study also demonstrated that carvacrol exhibits synergistic effect with chemotherapeutic drugs (5-FU and carboplatin). These initial findings implicated that natural compounds could reduce the toxic effects of chemotherapeutic drugs. Conclusion:: Therefore, this investigation affirms the anti-cancer potential of carvacrol against cervical cancer cells which could be an appendage in the prevention and treatment of cervical cancer.

LY294002 and Metformin Cooperatively Enhance the Inhibition of Growth and the Induction of Apoptosis of Ovarian Cancer Cells

2012 ◽  
Vol 22 (1) ◽  
pp. 15-22 ◽  
Author(s):  
Cuilan Li ◽  
Vincent Wing Sun Liu ◽  
David Wai Chan ◽  
Kwok Ming Yao ◽  
Hextan Yuen Sheung Ngan
Keyword(s):  
Cell Cycle ◽  
Ovarian Cancer ◽  
Flow Cytometry ◽  
Cell Growth ◽  
Cancer Cells ◽  
Cancer Cell ◽  
Ovarian Cancer Cell ◽  
Mtor Pathway ◽  
Ovarian Cancer Cells ◽  
Cancer Cell Growth

BackgroundThe phosphoinositide 3 kinase (PI3K)/v-akt murine thymoma viral oncogene homolog (AKT)/mammalian target of rapamycin (mTOR) pathway is frequently aberrantly activated in ovarian cancer and confers the chemoresistant phenotype of ovarian cancer cells. LY294002 (PI3K inhibitor) and metformin (5′-adenosine monophosphate [AMP]-activated protein kinase [AMPK] activator) are 2 drugs that were known to inhibit mTOR expression through the AKT-dependent and AKT-independent pathways, respectively. In this study, we explored the effectiveness of LY294002 and metformin in combination on inhibition of ovarian cancer cell growth.MethodsWestern blotting was used to detect the changes of PI3K/AKT/mTOR and AMPK/acetyl-CoA carboxylase (ACC) signaling activities, cell cycle control, and apoptosis. Cell growth was evaluated by cell proliferation, colony formation, and soft agar assays. Flow cytometry was used to study cell cycle distribution and cell death upon drug treatment.ResultsOur study showed that LY294002 and metformin in combination could simultaneously enhance the repression of the PI3K/AKT/mTOR pathway and the activation of the AMPK/ACC pathway. The downstream target of AKT and AMPK, mTOR, was cooperatively repressed when the drugs were used together. The cell cycle regulatory factors, p53, p27, and p21, were up-regulated. On the other hand, caspase 3 and poly (ADP-ribose) polymerase activities involved in apoptosis were also activated. Cell growth assays indicated that LY294002 and metformin could effectively inhibit ovarian cancer cell growth. Flow cytometry analysis showed that the treatment of the 2 drugs mentioned above induced cell cycle arrest at G1 phase and increased sub-G1 apoptotic cells.ConclusionThe combinational use of LY294002 and metformin can enhance inhibition of the growth and induction of the apoptosis of ovarian cancer cells. Our results may provide significant insight into the future therapeutic regimens in ovarian cancer.

scholarly journals Antibacterial activity of aquatic extract of Myrtus communis leaves against Periodontitis isolated bacteria

2021 ◽  
Vol 880 (1) ◽  
pp. 012047
Author(s):  
Eman Mubdir Nayf ◽  
Hamzah Abdulrhaman Salman
Keyword(s):  
Susceptibility Testing ◽  
Morphological Characterization ◽  
Diffusion Method ◽  
Potential Zone ◽  
Myrtus Communis ◽  
Zone Of Inhibition ◽  
Biochemical Tests ◽  
Gram Positive ◽  
Gram Positive Bacteria ◽  
Antibacterial Susceptibility

Abstract Myrtus communis is an evergreen plant that can survive stressful environments and high-temperature seasons. Treatment using green plants was the most trended in recent years. The present study aimed to evaluate the antibacterial effects of Myrtus communis leaves against bacteria isolated from periodontitis. Fifty samples were collected from periodontitis subjects in both genders (female 32 % and male 68 %). The isolates were diagnosed by morphological characterization and biochemical tests. M. communis leaves were identified, collected, and prepared for extraction. The plant leaves were extracted using distilled water. The antibacterial susceptibility testing was performed by the well diffusion method. Antibiotics susceptibility patterns were executed using the disc diffusion method. All the isolates belonged to gram-positive bacteria. Among the isolated bacteria, 20, 18, and 12 were Lactobacillus spp., Streptococcus spp., and Staphylococcus aureus, respectively. The antibacterial susceptibility testing of M. communis extract showed a potential zone of inhibition against all the tested bacteria. Of the different concentrations, 30 mg/ml showed the highest zone of inhibition, 18.2 mm, 19.50 mm, and 30.66 mm against Streptococcus spp., Staphylococcus spp., and Lactobacillus spp. Among the tested antibiotics, ciprofloxacin and chloramphenicol exhibited the highest zone of inhibition against the tested bacteria. The aquatic extract of M. communis leaves was found to be effective against gram-positive bacteria. Further studies are warranted to investigate the active bio-compounds.

scholarly journals Anti-proliferative mechanism of Huaier extract in human thyroid carcinoma cells

2020 ◽  
Author(s):  
Jingni He ◽  
Ying Zhang ◽  
Lidong Wang ◽  
Yifang Yu ◽  
Baiyu Yao ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Thyroid Cancer ◽  
Cancer Cells ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
Treated Group ◽  
G1 Phase ◽  
Thyroid Cancer Cells ◽  
Number Of Cells

Abstract BackgroundThyroid cancer is the most common endocrine tumor and typically has a good prognosis; however, some patients still present with local or distant metastases. Huaier is a traditional Chinese medicine reported as effective in treating certain types of tumor, but the effect of Huaier on thyroid cancer has not yet been reported. MethodsThe thyroid cancer cell lines, B-CPAP and C643, were treated with increasing concentrations of Huaier extract and the therapeutic effect was measured using a cell counting kit 8 (CCK-8) and flow cytometry. High-throughput sequencing was further performed to identify differentially expressed genes (DEGs) in Huaier-treated B-CPAP cells. Moreover, quantitative real-time PCR (RT-qPCR) was carried out to verify the selected RNAs. Finally, the dual luciferase detection kit was used to detect gene activity.ResultsProliferation of B-CPAP and C643 cells was significantly suppressed by treatment with Huaier extract in a concentration- and time-dependent manner. Huaier extract also induced cell cycle arrest and apoptosis according to flow cytometry (p < 0.05).High-throughput sequencing observed 7,979 significantly altered transcripts. Gene Ontology (GO) analysis showed that 270 genes were enriched in upregulated terms, while 171 genes were enriched in downregulated terms (p < 0.05). Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis indicated that there were 47 enriched pathways associated with DEGs (p < 0.05). The expression levels of chosen lncRNAs (SNHG7, MIR181A2HG, ILF3-AS1, and CTA-29F11.1) and their corresponding mRNAs (BBC3, CTSL, GADD45A, and DDIT3) were verified to be overexpressed in Huaier-treated B-CPAP cells by RT-qPCR (p < 0.05).Following transduction, the CCK-8 results showed that the proliferative capacity was increased in the shRNA group as compared with that in the Ctrl and Scr groups. According to flow cytometry, the number of cells in the G0/G1 phase was decreased in the shRNA group (p < 0.01) and the apoptosis rate was lower (p < 0.05). The shRNA-treated group had significantly reduced Huaier-induced apoptosis as compared with the Scr-treated group (p < 0.05). Moreover, the number of cells in the G0/G1 phase in the shRNA-treated group was significantly lower than that in the Scr-treated group (p < 0.05). The results of the dual luciferase reporter gene experiment showed that the activity in the GADD45A WT + miR-301a-3p(+) group was significantly reduced as compared with that in the GADD45A WT + miR-301a-3p(+) NC group (p < 0.01). Further, the activity in the ILF3-AS1 WT + miR-301a-3p(+) group was significantly lower than that in the ILF3-AS1 WT + miR-301a-3p(+) NC group (p < 0.05).ConclusionsThe present study demonstrates that Huaier extract inhibits the proliferation of thyroid cancer cells via changes in the expression levels of a multitude of genes. In particular, a decrease in GADD45A expression enhances the proliferative ability of thyroid cancer cells, the levels of which can be increased by Huaier treatment, thus regulating the cell cycle and apoptosis. Huaier can inhibit the proliferation of thyroid cancer cells through the ILF3-AS1/hsa-miR-301a-3p/GADD45A ceRNA axis.

Evaluation of Cytotoxic Mechanisms of Clinacanthus Nutans Extracts in Cancer Cells

2020 ◽  
Vol 10 ◽  
Author(s):  
Nurul Atiqah Sulaiman ◽  
Rajan Rajabalaya ◽  
Shirley Huan Fang Lee ◽  
Ya Chee Lim ◽  
Wei Hon Lim ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Cycle ◽  
Flow Cytometry ◽  
Cell Death ◽  
Cancer Cells ◽  
Cancer Cell ◽  
Annexin V ◽  
Cell Cycle Profile ◽  
Clinacanthus Nutans ◽  
Mcf 7

Background: Commercially available Clinacanthus nutans (Burm.F) Lindau (Acanthaceae) (CN) leaf preparations are gaining attention as an alternative cancer treatment, particularly in South East Asia. Multiple studies have suggested that CN has potential anticancer activities; however, the mechanism of these activities has remained elusive. Objective: This study evaluated the cytotoxic mechanisms of CN extracts in cancer cells. Methods: CN extracts were prepared from either fresh or dried leaves, using different solvents. Cytotoxicity of CN extracts were tested on the A549 (lung cancer), HeLa (cervical cancer), MCF-7 (breast cancer) and MDA-MB-231 (breast cancer) cell lines using the MTT assay. Flow cytometry was used to assess changes in the cell cycle profile, while Western blotting was used to examine microtubule stability. Finally, the mode of cell death was investigated using the Annexin V-FITC Apoptosis Detection Kit. Results: Aqueous Fresh (AQF) extract was prepared to simulate the ethno-medicinal use of CN, and reduced cell viability of MCF-7 cells with IC50 = 1.71 mg/mL. Some CN extracts have the ability to inhibit the proliferation of four different cancer cell lines after a 24 hour treatment. Annexin V assay results shows that acetone extracts of CN induced increments in percentage of apoptotic cell death. However, flow cytometry results show that cancer cell cycle profile were not affected. Similarly, immunoblotting results also indicate that microtubule dynamics in MCF-7 cells were not altered. However, the aqueous extract, prepared to simulate the current ethnomedicinal use of CN leaves in cancer treatment, did not significantly inhibit cancer cell proliferation with IC50 = 1.71 mg/mL. Conclusions: This study was the first to show that microtubules in cancer cells remain dynamic after treatment with CN extracts, effectively ruling interference of microtubule dynamics as the mode of cell death. AMD extract showed the highest effects MCF-7 cell proliferation.

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