scholarly journals P04.05 The Effects and Mechanism Involved in Paclitaxel combined with Shikonin on U87 Glioma Cells

2019 ◽  
Vol 21 (Supplement_3) ◽  
pp. iii29-iii29
Author(s):  
J SHI
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Cell Migration ◽  
Inhibitory Effect ◽  
Transwell Assay ◽  
Significant Inhibitory Effect ◽  
Rna Content ◽  
Paclitaxel Group ◽  
U87 Cells

Abstract BACKGROUND Glioma is the most common intracranial primary tumor,chemotherapy seems to be the main treatment for brain glioma.Chemotherapy is currently widely used in clinical treatment of gliomas,however,drug-resistant always appears,so combination of chemotherapy has became an inevitable choice. Paclitaxel and shikonin are stable and efficient chemotherapy drugs which can promote apoptosis and ininhibite the growth of tumor.Recent research reports,paclitaxel and shikonin can respectively activate and inhibit the expression of p-ERK.The purpose of this study was to investigate whether we can inhibite the activation of p-ERK caused by paclitaxel to enhance the effect of paclitaxel,furthermore use the combination of the two drugs to reach a better effect of chemotherapy. MATERIAL AND METHODS 1.Cell Culture2. Drugs Administration.3. Growth Inhibition Assays (CCK-8) .4.Scratch test to observe cell migration 5.Transwell assay for the determination of cell migration 6.Transwell assay for the determination of cell invasion 7. Annexin V-FITC and PI Assay for Flow Cytometry to Detect Apoptosis 8. PI Assay for Flow Cytometry to Detect Cell Cycle 9. Western Blot to Detect the Expression of P-ERK/ERK 10. Statistical Analysis. RESULTS 1.The results tested by CCK-8 showed these two drugs have synergic effects with a CDI equals to 0.72 which means these two drugs are significant synergistic.2.Compared with paclitaxel group or shikonin group,the 1/2paclitaxel+1/2shikonin group had a significant inhibitory effect upon the migration of U87 cells.3.Compared with paclitaxel group and shikonin group,the 1/2paclitaxel+1/2shikonin group had a significant inhibitory effect upon the invation of U87 cells.4.Compared with paclitaxel group and shikonin group,the 1/2paclitaxel+1/2shikonin group had a significant promoting effect upon inducing apoptosis of U87 cells.5. The 1/2paclitaxel+1/2shikonin group significantly improved the apoptosis rate of U87 cells,the RNA content of the G1-phase was significantly increased in shikonin group, the RNA content of the G2- phase significantly increased in paclitaxel group and 1/2paclitaxel+1/2shikonin group.6.Compared with control group,the rate of p-ERK/ERK was increased in paclitaxel group(P<0.05),while the rate of p-ERK/ERK was significantly decreased in the shikonin group and 1/2paclitaxel+1/2shikonin group(P<0.05). CONCLUSION 1. Compared with paclitaxel group and shikonin group,the combination group with half dosage can make synergic effects and significantly inhibit the proliferation,migration and invation ability of U87 cells,besides promote apoptosis of U87 cells and block U87 cell cycle.2. Paclitaxel combined with shikonin could decrease the expression of p-ERK/ERK,which may be involved in the synergic effects of these two drugs.

Targeted Modulation of FAK/PI3K/PDK1/AKT and FAK/p53 Pathways by Cucurbitacin B for the Antiproliferation Effect Against Human Cholangiocarcinoma Cells

2020 ◽  
Vol 48 (06) ◽  
pp. 1475-1489
Author(s):  
Sirinapha Klungsaeng ◽  
Veerapol Kukongviriyapan ◽  
Auemduan Prawan ◽  
Sarinya Kongpetch ◽  
Laddawan Senggunprai
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Tyrosine Kinases ◽  
Molecular Mechanisms ◽  
Inhibitory Effect ◽  
Chemotherapeutic Agents ◽  
Cucurbitacin B ◽  
M Phase ◽  
Cell Cycle Disruption

Inadequate responses to traditional chemotherapeutic agents in cholangiocarcinoma (CCA) emphasize a requirement for new effective compounds for the treatment of this malignancy. This study aimed to investigate the antiproliferative property of cucurbitacin B on KKU-100 CCA cells. The determination of underlying molecular mechanisms was also carried out. The results revealed that cucurbitacin B suppressed growth and replicative ability to form colonies of CCA cells, suggesting the antiproliferative effect of this compound against the cells. Flow cytometry analysis demonstrated that the interfering effect of cucurbitacin B on the CCA cell cycle at the G2/M phase was accountable for its antiproliferation property. Accompanied with cell cycle disruption, cucurbitacin B altered the expression of proteins involved in the G2/M phase transition including downregulation of cyclin A, cyclin D1, and cdc25A, and upregulation of p21. Additional molecular studies demonstrated that cucurbitacin B suppressed the activation of focal adhesion kinase (FAK) which consequently resulted in inhibition of its kinase-dependent and kinase-independent downstream targets contributing to the regulation of cell proliferation including PI3K/PDK1/AKT and p53 proteins. In this study, the transient knockdown of FAK using siRNA was employed to ascertain the role of FAK in CCA cell proliferation. Finally, the effect of cucurbitacin B on upstream receptor tyrosine kinases regulating FAK activation was elucidated. The results showed that the inhibitory effect of cucurbitacin B on FAK activation in CCA cells is mediated via interference of EGFR and HER2 expression. Collectively, cucurbitacin B might be a promising drug for CCA treatment by targeting FAK protein.

scholarly journals Growth Effects of Some Platinum(II) Complexes with Sulfur-Containing Carrier Ligands on MCF7 Human Breast Cancer Cell Line upon Simultaneous Administration with Taxol

2002 ◽  
Vol 9 (1-2) ◽  
pp. 33-43 ◽  
Author(s):  
Gordana Bogdanović ◽  
Vesna Kojić ◽  
Tatjana Srdić ◽  
Dimitar Jakimov ◽  
Miloš I. Djuran ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Recovery Period ◽  
Cancer Cell Line ◽  
Inhibitory Effect ◽  
Mcf7 Cells ◽  
Cisplatin Analogues ◽  
Ic50 Values ◽  
G2m Phase ◽  
Induced Apoptosis

The platinum (II)complexes, cis-[PtCl2(CH3SCH2CH2SCH3)] (Pt1), cis-[PtCl2(dmso)2] (dmso is dimethylsulfoxide; Pt2) and cis-[PtCl2(NH3)2] (cisplatin), and taxol (T) have been tested at different equimolar concentrations. Cells were exposed to complexes for 2 h and left to recover in fresh medium for 24, 48 or 72 h. Growth inhibition was measured by tetrazolium WST1 assay Analyses of the cell cycle, and apoptosis were performed by flow cytometry, at the same exposure times. The IC50 value of each platinum(II) complex as well as combination index (CI; platinum(II) complex + taxol) for various cytotoxicity levels were determined by median effects analysis.MCF7 cells were found to be sensitive to both Pt1 and Pt2 complexe These cisplatin analogues influenced the cell growth more effectively as compared to cisplatin. Cytotoxic effect was concentration and time-dependent. Profound growth inhibitory effect was observed for Pt1 complex, across all its concentrations at all recovery periods. A plateau effect was achieved three days after treatment at Pt1 concentrations ≤ 1 μM. Pt2, however, decreased MCF7 cells survival only for the first 24 h ranging between 50-55%. Pt2 cytotoxicity sharply decreased thereafter, approaching 2 h - treatment cytotoxicity level. The median IC50 values for Pt1 and Pt2 were similar (0.337 and 0.3051 μM, respectively) but only for the first 24 h. The IC50 values for Pt1 strongly depend on the recovery period. On simultaneos exposure of cells to taxol and platinum(II) complexes no consistent effect was found. The Cls for combinations of taxol with Pt1 or Pt2 revealed cytotoxic effects that were in most Cases synergistic (Pt1) or less than addtiive (Pt2). Flow cytometry analysis has shown that each platinum(II) complex induced apoptosis in MCF7 cells. The level of apoptosis correlated with cytotoxicity level for the range concentrations. Both cisplatin analogues, at IC50 concentrations, increased the number of MCF7 cells in G0G1 phase of cell cycle. Pt2-treated cells remained arrested in G0G1 phase up to 72 h after treatment. Combination of Pt2 and taxol caused further arrest of cells in G0G1 phase (24 h) in parallel with strong decrement of G2M phase cells.

Myricanol-9-acetate, a novel naturally occurring derivative of myricanol, induces ROS-dependent mitochondrial-mediated Apoptosis in MCF-7 cancer cells

2021 ◽  
Vol 21 ◽  
Author(s):  
Gazanfar Ahmad ◽  
Sameer Ahmad Mir ◽  
Loveleena Kour Anand ◽  
Faheem Hyder Pottoo ◽  
Neerupma Dhiman ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Cell Line ◽  
Ros Generation ◽  
Cycle Arrest ◽  
Naturally Occurring ◽  
Induced Apoptosis ◽  
Dose Dependent ◽  
Mcf 7

Background: Low therapeutic efficacy and drug-induced systemic toxicity of currently used anti-cancerous chemotherapeutic agents are major compelling factors for finding out clinically efficient molecules with high efficiency and less toxicity. Objective: The current research work was undertaken to evaluate the anticancer potential of Myricanol-9-acetate (MA), a novel naturally occurring derivative of myricanol. Methods: MCF-7, MiaPaCa-2, and HCT 116 were used for cytotoxicity determination of the MA and ML (Myricanol) by MTT assay. The mechanistic study involved the determination of cell cycle arrest, ΔΨm loss, ROS generation, western blot assay, and flow cytometry by reported methods on MCF-7 cells. Results: MA exhibited anticancer activity against all three cell lines; however, the molecule was found most active against the MCF-7 cell line. We observed IC50 20μM with MA treatment as compared to the IC50 of 42 μM for myricanol treatment. Detailed mechanistic studies revealed that MA induced apoptosis on MCF-7 cell line through ROS generation; a dose-dependent drop in mitochondrial membrane potential was found to be associated with cell cycle arrest at G0/G1 phase. Our results further demonstrated down-regulation of Bcl2 and activation of the caspase cascade as the events involved in the MA-induced apoptosis. Flow cytometry results indicated an increase in early and late apoptotic population in a dose-dependent manner with an apoptotic population of about 20% at 30 μM of MA, thus supporting our results. Conclusion: Present findings thus suggest that MA might serve as a promising novel drug candidate having high scope for further evaluation in preclinical and clinical studies.

scholarly journals ST09, A Novel Curcumin Derivative, Blocks Cell Migration by Inhibiting Matrix Metalloproteases in Breast Cancer Cells and Inhibits Tumor Progression in EAC Mouse Tumor Models

Molecules ◽  
2020 ◽  
Vol 25 (19) ◽  
pp. 4499
Author(s):  
Snehal Nirgude ◽  
Raghunandan Mahadeva ◽  
Jinsha Koroth ◽  
Sujeet Kumar ◽  
Kothanahally S. Sharath Kumar ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Cycle ◽  
Flow Cytometry ◽  
Cell Death ◽  
Cell Migration ◽  
Drug Toxicity ◽  
Mouse Tumor ◽  
Curcumin Derivative

Purpose: Curcumin is known for its anticancer and migrastatic activity in various cancers, including breast cancer. Newer curcumin derivatives are being explored to overcome limitations of curcumin like low bioavailability, stability, and side effects due to its higher dose. In this study, the synthesis of ST09, a novel curcumin derivative, and its antiproliferative, cytotoxic, and migrastatic properties have been explored both in vitro and in vivo. Methods: After ST09 synthesis, anticancer activity was studied by performing standard cytotoxicity assays namely, lactate dehydrogenase (LDH) release assay, 3-(4, 5-dimethylthiazol-2-yl)-2–5-diphenyletrazolium bromide (MTT), and trypan blue exclusion assay. Annexin-FITC, cell cycle analysis using flow cytometry, and Western blotting were performed to elucidate cell death mechanisms. The effect on the inhibition of cell migration was studied by transwell migration assay. An EAC (Ehrlich Ascites carcinoma) induced mouse tumor model was used to study the effect of ST09 on tumor regression. Drug toxicity was measured using aspartate aminotransferase (AST), alanine aminotransferase (ALT), blood urea nitrogen (BUN), and flow-cytometry based lymphocyte count. Histological analysis was performed for assessment of any tissue injury post ST09 treatment. Results: ST09 shows an approximate 100-fold higher potency than curcumin, its parent compound, on breast tumor cell lines MCF-7 and MDA-MB231. ST09 arrests the cell cycle in a cell type-specific manner and induces an intrinsic apoptotic pathway both in vitro and in vivo. ST09 inhibits migration by downregulating matrix metalloprotease 1,2 (MMP1,2) and Vimentin. In vivo, ST09 administration led to decreased tumor volume in a mouse allograft model by boosting immunity with no significant drug toxicity. Conclusion: ST09 exhibits antiproliferative and cytotoxic activity at nanomolar concentrations. It induces cell death by activation of the intrinsic pathway of apoptosis both in vitro and in vivo. It also inhibits migration and invasion. This study provides evidence that ST09 can potentially be developed as a novel antitumor drug candidate for highly metastatic and aggressive breast cancer.

Mechanism Underlying the Inhibitory Effect of Bio-synthesized Silver Nanoparticle on TNF-α induced NF- κB Nuclear Translocation in Prostate Cancer Cells

2021 ◽  
Vol 14 (2) ◽  
pp. 5396-5405
Author(s):  
Rajathi K ◽  
Leneeygreen K.B ◽  
Suja S
Keyword(s):  
Prostate Cancer ◽  
Cell Cycle ◽  
Flow Cytometry ◽  
Cell Death ◽  
Silver Nanoparticle ◽  
Nuclear Translocation ◽  
Inhibitory Effect ◽  
Human Prostate Cancer ◽  
Tnf Α ◽  
M Phase

Apoptosis, a physiological mechanism of highly orchestrated cell death, can be initiated by extracellular and intracellular mechanisms that trigger a complex machinery of proapoptotic proteases and mitochondrial changes, leading to the activation of specific endonucleases and DNA fragmentation. The present study was undertaken to elucidate a mechanism underlying the inhibitory effect of biosynthesised silver nanoparticle on TNF-α induced NF-κB nuclear translocation in prostate cancer PC- 3 cells. The  cell cycle analysis of Prostate cancer PC-3 cells was examined by flow cytometry by using annexin V-FITC/PI staining. Effect of silver nanoparticles in oxidative stress ROS, Effect of biosynthesized silver nanoparticle on apoptosis in human prostate cancer cell line and apoptotic induction of TNF-α and NF-κB expression was studied by Flow cytometry in Prostate cancer PC-3 cell line. From the results it was observed that biosynthesized silver nanoparticle inhibits the cellular growth of human prostate cancer PC-3 cells and induces apoptosis. The ROS levels generated in response to silver nanoparticles were significantly higher in treated PC-3 cells than the control. The result indicates that cell death is mediated by ROS production, which might alter the cellular redox status, and it is a potential reason for cell death. Apoptosis of the silver nanoparticle treated PC-3 cells was accompanied by a reduction in the percentage of cells in G0/G1 phase and an increase in the percentage of G2/M phase cells, indicating cell cycle arrest at G2/M phase, and transcription factor NF-κB plays an essential role in inflammation and cancer. The activation of NF-κB in response to inflammatory cytokine such as TNF-α promotes nuclear migration to enable DNA-binding activity and facilitate target genes expression.

scholarly journals The Inhibitory Effect of Curcumin on Hypoxia Inducer Factors (Hifs) as a Regulatory Factor in the Growth of Tumor Cells in Breast Cancer Stem-Like Cells

Drug Research ◽  
2020 ◽  
Vol 70 (11) ◽  
pp. 512-518
Author(s):  
Mehrnaz Asadi Sarighieh ◽  
Vahideh Montazeri ◽  
Amir Shadboorestan ◽  
Mohammad Hossein Ghahremani ◽  
Seyed Nasser Ostad
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Chemotherapy Resistance ◽  
Antitumor Agent ◽  
Inhibitory Effect ◽  
Inhibitory Effects ◽  
Hypoxic Conditions ◽  
Western Blot Method ◽  
Cancer Agent ◽  
Mcf 7

AbstractHypoxia in the microenvironment is related to chemotherapy resistance, tumor progression, and metastasis. Curcumin, as a phenolic compound extracted from the turmeric, has been used as an anti-cancer agent with low toxicity in recent years. Since curcumin has inhibitory activities against hypoxia-inducible factors (HIFs) in several cancers, this study was conducted to examine the effect of curcumin on MCF-7 cells and cancer stem-like cells (CS-LCs) under hypoxic and normoxic conditions. CS-LCs were isolated from MCF-7 cells using the magnet activated cell sorting (MACS) method based on CD44 +/ CD24 - surface markers. The effects of curcumin on the viability of MCF-7 cells and CS-LCs were examined in hypoxic and normoxic conditions using the MTT test. The effects of curcumin on apoptosis and cell cycle of CS-LCs and MCF-7 cells were analyzed using flow cytometry. Moreover, the inhibitory effects of curcumin on the levels of HIF-1 and HIF-2α protein in CS-LCs were investigated using the western blot method. Early apoptosis occurred in CSC-LCs more than MCF-7 cells under hypoxic conditions. Flow cytometry assay showed that curcumin caused cell cycle arrest of CSC-LCs and MCF-7 at the G2/M phase under hypoxic conditions while under normoxic conditions, arrest occurred at the G0/G1 phase in MCF-7 cells and at S and G2/M phases in CS-LCs. Based on the results, the curcumin inhibited the expression of HIF-1 by degrading ARNT in CS-LCs.In conclusion, curcumin has inhibitory effects on MCF- 7 cells and CS- LCs and thus may be used as an antitumor agent.

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