scholarly journals Efek Sinergis Kombinasi Ekstrak Air Akar Batu (Gerrardanthus Macrorhizus) dengan Doxorubicin pada Sel Kanker Payudara T47D

2018 ◽  
Vol 46 (3) ◽  
pp. 183-190
Author(s):  
Sari Haryanti
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Cytotoxic Effect ◽  
Clinical Results ◽  
Morphological Alteration ◽  
T47d Cells ◽  
Natural Agents ◽  
Chemotherapy Agent ◽  
M Phase ◽  
Apoptotic Induction

Doxorubicin is one of the most widely used chemotherapy agents in cancer therapy. The clinical results of doxorubicin are limited by cardiotoxic side effects which correlates with dose. Combination of doxorubicin with natural agents is a promising strategy for reducing its therapeutic doses as well as decreasing cardiotoxic effects. This study was conducted to investigate cytotoxic activity of caudex Gerrardanthus macrorhizus aqueous extract and its combination with doxorubicin on T47D cells. Dried caudex powder was extracted by infusion method, evaporated in oven 400C to get dried extract (GM). The MTT assay was performed to determine cytotoxic effect, either alone or in combination. Flow cytometry is used to observe cell cycle profile and apoptotic induction. GM alone did not exhibit cytotoxic effects but caused morphological alteration in T47D cells. Nevertheless, GM 40 μg/mL was able to improve cytotoxic effect of doxorubicin to 51%. Combination of doxorubicin 3 nM and 40 μg/mL GMA inhibited cell cycle in G2/M phase. This combination also resulted apoptotic induction, compared to untreated cell and each single treatment. Based on the results, caudex G. macrorhizus is potential to be further investigated as a co-chemotherapy agent with doxorubicin. Abstrak Doxorubicin adalah salah satu agen kemoterapi yang digunakan secara luas dalam terapi kanker. Hasil klinis doxorubicin dibatasi oleh kardiotoksisitas yang berkorelasi dengan besaran dosis. Kombinasi doxorubicin dengan bahan alam merupakan strategi yang menjanjikan untuk mengurangi dosis terapetik doxorubicin sehingga dapat menurunkan efek samping kardiotoksik. Penelitian ini dilakukan untuk mengkaji aktivitas sitotoksik ekstrak air  caudex Gerrardanthus macrorhizus dan kombinasinya dengan doxorubicin pada sel T47D. Serbuk caudex G. macrorhizus (GM) kering diekstraksi dengan metode infusa, kemudian diuapkan dalam oven 400C. Uji MTT dilakukan untuk menguji efek sitotoksik ekstrak, baik tunggal maupun kombinasi dengan doxorubicin. Flow cytometry digunakan untuk mengetahui profil siklus sel dan induksi apoptosis. Hasil penelitian menunjukkan ekstrak tunggal tidak memberikan efek sitotoksik namun mengakibatkan perubahan morfologi pada sel T47D. Ekstrak GM 40 μg/mL mampu meningkatkan efek sitotoksik doxorubicin 3 nM hingga 51%. Kombinasi doxorubicin 3 nM dan ekstrak 40 μg/mL menghambat siklus sel pada fase G2/M dan meningkatkan induksi apoptosis, dibandingkan kontrol sel dan masing-masing perlakuan tunggalnya. Berdasarkan hasil penelitian ini, caudex G. macrorhizus berpotensi untuk diteliti lebih lanjut sebagai agen ko-kemoterapi dengan doxorubicin.  

Apoptosis Was Induced by Casticin in Acute Myelocytic Leukemia Cells

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 3991-3991
Author(s):  
Jie Jin ◽  
Jia-Kun Shen ◽  
Hua-ping Du ◽  
Min Yang ◽  
Yun-Gui Wang
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Cell Death ◽  
Leukemic Cell ◽  
K562 Cells ◽  
Acute Myelocytic Leukemia ◽  
Myelocytic Leukemia ◽  
Cycle Arrest ◽  
M Phase ◽  
Ic50 Values

Abstract Casticin, a component from Vitex rotundifolia wich was widely used as an anti-inflammatory agent in Chinese traditional medicine, was reported to have anti-tumor activities in lung cancer and breast cancer. There are yet no reports on roles against acute myelocytic leukemia (AML). This study aims to elucidate the anti-leukemic activity of casticin on AML cells. We investigated the efficient efficacy and the mechanisms by which casticin triggers cell death in AML cells by analyzing cell cycle perturbations, apoptosis-related marker expression. Cell viability was measured by MTT method; apoptosis and cell cycle arrest were determined by flow cytometry and AV-PI assay. Western blot was performed to measure the apoptosis-related marker. Concentration-dependant cell deaths were observed in AML cell lines including K562, U937 and THP-1, with IC50 values of 24h (hours) being 47.4μM, 67.8μM and 61.7μM, respectively. Time-dependant cell deaths were also observed. At the concentration of 20μM casticin, 45.7%, 76.1% and 80.9% of K562 cells were inhibited at 24h, 48h and 72h, respectively; 24.7%, 30% and 61% of U937 cells were inhibited at 24h, 48h and 72h, respectively; while for THP-1, 29%, 41.8% and 53.9% were inhibited at 24h, 48h and 72h, respectively. Apoptosis was found using AV-PI staining by flow cytometry analysis. We observed an obvious G2/M phase increase prolongation in casticin treated K562 cells. BThe distribitions of G2/M phase were 2.9%, 33.6%, 75.3%, 54.9%, 29.7% and 27.0% in K562 cells after treated by 20μM casticin for 0h, 6h, 12h, 24h, 36h and 48h, respectively. Furthermore, apoptosis-related proteins, PARP and caspase 3, were cleaved in casticin treated K562 cells. Taken together, these results demonstrated that casticin can induce leukemic cell death through apoptosis, suggesting that casticin could be a promising therapeutic agent against acute myeloid leukemia.

Aptamer TY04 Inhibits Multiple Myeloma Cell Growth Via Cell Cycle Arrest

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 5390-5390
Author(s):  
Jing Liu ◽  
Hong-Juan Dai ◽  
Bian-Ying Ma ◽  
Jian-Hui Song ◽  
Hui-yong Chen ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Multiple Myeloma ◽  
Flow Cytometry ◽  
Cell Growth ◽  
Cell Lines ◽  
Myeloma Cell ◽  
Multiple Myeloma Cell ◽  
Cycle Arrest ◽  
M Phase ◽  
Target Molecules

Abstract Multiple myeloma (MM), also known as plasma cell myeloma, is characterized by accumulation of clonal plasma cells in the bone marrow and overproduction of monoclonal immunoglobulin (Ig) in the blood or urine. MM accounts for approximately 10% of all hematologic malignancies. Despite recent advances in the understanding and treatment of this disease, MM remains an incurable disease in the vast majority. With conventional chemotherapy, the 5-year median survival rate for MM patients is approximately 25%. Aptamers are single-stranded RNA or DNA sequences that bind to target molecules with high affinity and specificity. Compared with antibodies, aptamers have unique advantages including easy chemical synthesis and modification, low toxicity, lack of immunogenicity, and rapid tissue penetration, Based on these advantages, aptamers show great potential for therapeutic application. The aptamer TY04 is a single-stranded DNA (ssDNA) generated by a method named cell-based systematic evolution of ligands by exponential enrichment (cell-SELEX), We found TY04 strongly inhibited the growth of multiple myeloma cell lines including MM1.S, NCI-H929, KM3 and OPM2,The concentration of TY04 to inhibit 50% cell growth (IC50) on MM1.S was 3.89 μM. In contrast, TY04 had no effect on the growth of non-tumor cell lines — immortal B lymphoblastoid cell lines. Next, we used MM1.S cell line as the model to study the mechanism of TY04 anti- multiple myeloma. Flow cytometry analysis showed that TY04 with the sequence specifically bind to MM1.S cells when compared with unselected ssDNA library control. To investigate whether the target molecules of TY04 are membrane proteins on cell surface, MM1.S cells were treated with trypsin and proteinase k for 2 or 10 minutes before incubation with TY04. The result revealed that TY04 lost partly recognition ability on treated cells, indicating that the target molecules were most likely membrane proteins. Furthermore, we evaluated the cell cycle distribution of MM1.S after TY04 treatment. We found that TY04 significantly caused cell-cycle arrest in G2/M phase. The percentage of G2/M phase cells increased from 30.1±1.56 to 53.2±6.36. To identify the underlying molecular mechanism, G2/M-related proteins were assayed by flow cytometry. Following TY04 treatment, a concomitant inhibition of ERK1/2, cyclin B, CDK1 and γ-tubulin expression occurred. Meanwhile, human cell cycle PCR array was used to analyze the expression of 84 genes key to cell cycle regulation in TY04-treated MM1.S cells. Our results indicated that aptamer TY04 decreased the genes expression of CCNB1, CCNB2, BIRC5, BRCA1 and CCNH, which were involved in the progress of G2/M phase. All these results are significant in that they provide a framework for further exploring the use of TY04 as a novel anti-multiple myeloma agent. Disclosures: No relevant conflicts of interest to declare.

Protein kinase D inhibitor CRT0066101 suppresses bladder cancer growth in vitro and in vivo through induction of cell cycle arrest at the G2-M phase.

2017 ◽  
Vol 35 (15_suppl) ◽  
pp. e16024-e16024
Author(s):  
Qingdi Quentin Li ◽  
Iawen Hsu ◽  
Thomas Sanford ◽  
Reema S. Railkar ◽  
Piyush K. Agarwal
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Bladder Cancer ◽  
Cyclin B1 ◽  
Cancer Growth ◽  
Protein Kinase D ◽  
Bladder Cancer Cells ◽  
M Phase

e16024 Background: Protein Kinase D (PKD) is implicated in tumor growth, death, invasion, and progression. CRT0066101 is an inhibitor of PKD and has antitumor activity in several types of carcinomas. However, the effect and mechanism of CRT0066101 in bladder cancer remain unknown. Methods: The MTS assay was used to evaluate the ability of CRT0066101 to inhibit cellular proliferation in bladder cancer cells. Cell cycle was analyzed by flow cytometry. Protein expression and phosphorylation were assessed by western blotting. Results: We showed that CRT0066101 suppressed the proliferation and migration of 4 bladder cancer cell lines in vitro. We also demonstrated that CRT0066101 inhibited tumor growth in an in vivo mouse model of bladder cancer. To verify the role of PKD in bladder tumor, we found that PKD2 was highly expressed in 8 bladder cancer lines and that RNA interference-mediated silencing of the PKD2 gene dramatically reduced bladder cancer growth in vitro and in vivo, suggesting that the effect of the compound in bladder cancer is mediated through inhibition of PKD2. This notion was confirmed by demonstrating that the levels of PKD2 and phospho-PKD2 (Ser-876) were markedly decreased in CRT0066101-treated bladder cancer. In addition, our cell cycle analysis by flow cytometry revealed that CRT0066101 arrested bladder cancer cells at the G2-M phase. We further validated these data by immunoblotting showing that treatment of bladder carcinoma cells with CRT0066101 downregulated the expression of cyclin B1, cdc2 and cdc25C, but elevated the levels of p27kip1, gadd45a, chk1/2, and wee1. Finally, CRT0066101 was found to increase the phosphorylation of cdc2 and cdc25C, which lead to reduction in cdc2-cyclin B1 activity. Conclusions: These novel findings suggest that CRT0066101 inhibits bladder cancer growth through modulating the cell cycle G2 checkpoint and inducing cell cycle G2-M arrest, which lead to blockade of cell cycle progression. QQL and IH contributed equally to this work.

scholarly journals Paris polyphylla 26 triggers G2/M phase arrest and induces apoptosis in HepG2 cells via inhibition of the Akt signaling pathway

2019 ◽  
Vol 47 (4) ◽  
pp. 1685-1695
Author(s):  
Qiang Li ◽  
Zifan He ◽  
Jiming Liu ◽  
Jianlong Wu ◽  
Guixiang Tan ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Signaling Pathway ◽  
Hepg2 Cells ◽  
Akt Signaling ◽  
Akt Signaling Pathway ◽  
Phase Arrest ◽  
Paris Polyphylla ◽  
M Phase ◽  
Related Proteins

Objectives Paris polyphylla 26 (PP-26) is a monomer purified from Paris polyphylla, which has traditionally been used as an antimicrobial, hemostatic, and anticancer agent in China. The anti-proliferation effect and underlying molecular mechanism of PP-26 were investigated in vitro. Methods The effects of PP-26 on various tumor cells were detected by MTT assay. PP-26-affected cell cycle and cell cycle-related proteins in HepG2 cells were detected by flow cytometry and western blotting, respectively. Apoptosis in response to PP-26 was assessed by Hoechst 33258 staining and flow cytometry. PP-26-affected apoptosis-related proteins and Akt signaling were detected by western blotting. The inhibitory effect of PP-26 on HepG2 cells, when combined with 5-fluorouracil (5-FU), was also assessed. Results PP-26 inhibited proliferation of HepG2 cells in a dose-dependent manner by triggering G2/M-phase arrest. Moreover, PP-26 induced apoptosis of HepG2 cells. Expression levels of apoptosis proteins caspase 9, caspase 3, PARP, Bcl-2, Bcl-xL, and Mcl-1 were downregulated, while the expression level of apoptosis protein Bax was upregulated. Expression levels of p-Akt, p-GSK-3β, and p-Foxo3 were downregulated. Combination with PP-26 enhanced 5-FU inhibition of HepG2 cell proliferation. Conclusions PP-26 triggers G2/M-phase arrest and induces apoptosis in HepG2 cells via inhibition of the Akt signaling pathway.

Cell cycle delay and apoptosis are induced by high salt and urea in renal medullary cells

2000 ◽  
Vol 278 (2) ◽  
pp. F209-F218 ◽  
Author(s):  
L. Michea ◽  
D. R. Ferguson ◽  
E. M. Peters ◽  
P. M. Andrews ◽  
M. R. Kirby ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Morphological Changes ◽  
Collecting Duct ◽  
S Phase ◽  
Electron Microscopic ◽  
Cell Cycle Delay ◽  
M Phase

We investigated the effects of hyperosmolality on survival and proliferation of subconfluent cultures of mIMCD3 mouse renal collecting duct cells. High NaCl and/or urea (but not glycerol) reduces the number of viable cells, as measured with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT). Raising osmolality from a normal level (300 mosmol/kg) to 550–1,000 mosmol/kg by adding NaCl and/or urea greatly increases the proportion of cells in the G2M phase of the cell cycle within 8 h, as measured by flow cytometry. Up to 600 mosmol/kg the effect is only transient, and by 12 h at 550 mosmol/kg the effect reverses and most cells are in G1. Flow cytometry with 5-bromodeoxyuridine (BrdU) pulse-chase demonstrates that movement through the S phase of the cell cycle slows, depending on the concentrations of NaCl and/or urea, and that the duration of G2M increases greatly (from 2.5 h at 300 mosmol/kg to more than 16 h at the higher osmolalities). Addition of NaCl and/or urea to total osmolality of 550 mosmol/kg or more also induces apoptosis, as demonstrated by characteristic electron microscopic morphological changes, appearance of a subdiploid peak in flow cytometry, and caspase-3 activation. The number of cells with subdiploid DNA and activated caspase-3 peaks at 8–12 h. Caspase-3 activation occurs in all phases of the cell cycle, but to a disproportionate degree in G0/G1 and S phases. We conclude that elevated NaCl and/or urea reduces the number of proliferating mIMCD3 cells by slowing the transit through the S phase, by cell cycle delay in the G2M and G1, and by inducing apoptotic cell death.

scholarly journals Implication of Lactucopicrin in Autophagy, Cell Cycle Arrest and Oxidative Stress to Inhibit U87Mg Glioblastoma Cell Growth

Molecules ◽  
2020 ◽  
Vol 25 (24) ◽  
pp. 5843
Author(s):  
Rossella Rotondo ◽  
Maria Antonietta Oliva ◽  
Sabrina Staffieri ◽  
Salvatore Castaldo ◽  
Felice Giangaspero ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cell Cycle ◽  
Cell Growth ◽  
Cell Cycle Arrest ◽  
Cytotoxic Effect ◽  
Cycle Arrest ◽  
M Phase ◽  
Clonogenic Potential ◽  
Pre Treatment ◽  
Cell Growth And Viability

In this study, we propose lactucopicrin (LCTP), a natural sesquiterpene lactone from Lactucavirosa, as a molecule able to control the growth of glioblastoma continuous cell line U87Mg. The IC50 of U87Mg against LCTP revealed a strong cytotoxic effect. Daily administration of LCTP showed a dose and time-dependent reduction of GBM cell growth and viability, also confirmed by inhibition of clonogenic potential and mobility of U87Mg cells. LCTP activated autophagy in U87Mg cells and decreased the phosphorylation of proliferative signals pAKT and pERK. LCTP also induced the cell cycle arrest in G2/M phase, confirmed by decrease of CDK2 protein and increase of p53 and p21. LCTP stimulated apoptosis as evidenced by reduction of procaspase 6 and the increase of the cleaved/full-length PARP ratio. The pre-treatment of U87Mg cells with ROS scavenger N-acetylcysteine (NAC), which reversed its cytotoxic effect, showed the involvement of LCTP in oxidative stress. Finally, LCTP strongly enhanced the sensitivity of U87Mg cells to canonical therapy Temozolomide (TMZ) and synergized with this drug. Altogether, the growth inhibition of U87Mg GBM cells induced by LCTP is the result of several synergic mechanisms, which makes LCTP a promising adjuvant therapy for this complex pathology.

Malignant Melanoma in Children and Congenital Melanocytic Nevi: DNA Content and Cell Cycle Analysis by Flow Cytometry

2001 ◽  
Vol 4 (1) ◽  
pp. 73-81 ◽  
Author(s):  
Antonio Alvarez-Mendoza ◽  
Jorge Reyes-Esparza ◽  
Ramon Ruiz-Maldonado ◽  
Eduardo Lopez-Corella ◽  
Norma C. Juarez-Herrera
Keyword(s):  
Risk Factors ◽  
Cell Cycle ◽  
Flow Cytometry ◽  
Malignant Melanoma ◽  
Dna Content ◽  
Melanocytic Nevus ◽  
S Phase ◽  
Congenital Melanocytic Nevus ◽  
M Phase ◽  
Diploid Cells

Malignant melanoma (MM) in children, although a rare neoplasm, can occur within a preexisting congenital melanocytic nevus (CMN). All the potential risk factors for this phenomenon are not well known, but increases in S phase and G2 + M phase of cell cycle, DNA aneuploidy, and cell cycle abnormalities in precursor lesions might be among the risk factors. Using paraffin-embedded tissue, we performed a retrospective analysis of DNA content, aneuploidy, and cell cycle by flow cytometry. Two groups of patients were analyzed: 28 children with CMN who did not developed MM, and 6 patients who further developed MM. In this second group, three patients had four biopsies done before the appearance of MM and in two patients biopsies were done after the appearance of MM. All CMN not associated with MM exhibited diploid cells only, their S phase was 11.5% (± 3.8), and their G2 + M phase was 2.5% (± 2.2). Among those patients who developed MM, 3/6 had an S phase > 15.5 and a G2 + M phase > 2.3 prior to the appearance of MM. Two out of six patients had a tetraploid DNA when MM developed and died with a disseminated MM. They had an S phase > 15.5 and their G2 + M phase was > 2.5. We propose that evaluation of DNA content and cell cycle by flow cytometry is a useful method to supplement biopsy findings in children with CMN who have lesions suspicious of developing a MM.

scholarly journals Exposure of Murattal Al-Quran Audio Enhances Cisplatin Activity on Growth Inhibition and Cell Cycle Modulation on Hela Cells

2019 ◽  
Vol 10 (2) ◽  
pp. 71 ◽  
Author(s):  
Roihatul Mutiah ◽  
Muhammad Ragib Mustofa ◽  
Yen Yen Ari Indrawijaya ◽  
Abdul Hakim ◽  
Rahmi Annisa ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Music Therapy ◽  
Hela Cells ◽  
Supportive Therapy ◽  
Cancer Treatments ◽  
Mtt Method ◽  
Cell Cycle Modulation ◽  
M Phase ◽  
Induction Of Apoptosis

Cancer is a disease characterized by abnormal cell mechanisms. The development of alternative cancer treatments is still needed. One of them is the music therapy. The music therapy uses sound vibrations to improve healing. Al-Quran, the Holy book of muslims, recites with measured recitation produces beautiful tones. Reading and recitating of Al-Quran is an important form of worship for which a Muslim can expect reward and benefit in the Hereafter. Al-Fatihah is one of surahs in Al-Qur'an that is often read by Muslims and used as a prayer for healing. The purpose of this study was to determine the effect of cytotoxic activity and cell cycle modulation on Hela cells with exposure of murattal Al-Quran and cisplatin combination. Audio exposure murattal Al-Fatihah and its combination with cisplatin to HeLa cells were tested using the MTT method assay. Induction of apoptosis and modulation of cell cycle evaluated by flow cytometry method. Treatment used 30 minutes Audio Murattal (AM), Cisplatin 10 µg/mL (Cis), and the combination of AM + Cis caused a decrease in the viability of HeLa cells respectively 80.14%, 69.86%, and 64.32%. The results of flow cytometry explained that in treatment of AM there was inhibition in the G2-M phase and induction of apoptosis in the M5 phase. Whereas in treatment AM + Cis inhibition occurs in the S, G2-M phase, and induction of apoptosis in the M5 phase. Audio Murattal Al-Quran presents cytotoxic effects on HeLa cells and to provide a synergistic impact on cisplatin so that disclosure therapy murattal can be recommended for supporting therapy in the treatment of cancer (supportive therapy).Keywords: Al-Quran, Audio murottal, HeLa Cells, Cell Viability, Flow cytometry

Comparative analysis of cell cycle parameters in primary and recurrent soft tissue sarcomas.

2017 ◽  
Vol 35 (15_suppl) ◽  
pp. e22538-e22538
Author(s):  
Inna Arnoldovna Novikova ◽  
Evgeniya M. Nepomnyashchaya ◽  
Timur Aliev ◽  
Elena Yurievna Zlatnik ◽  
Olesya N. Selyutina ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Comparative Analysis ◽  
Soft Tissue ◽  
Dna Content ◽  
Soft Tissue Sarcomas ◽  
Disease Stage ◽  
Primary Tumors ◽  
Recurrent Tumors ◽  
M Phase

e22538 Background: The purpose of the study was to determine DNA content and distribution of cells in cell cycle phases by flow cytometry in patients with primary and recurrent soft tissue sarcomas. Methods: 60 patients with soft tissue sarcomas (STS) were recruited: 30 with primary tumors and 30 with recurrent ones. Mean age of patients with primary STS was 56±5.4 years, with recurrent STS – 55±6.7 years. DNA content was determined using the BD Facs Cantoo II flow cytometer with CycleTEST PLUS DNA Reagent Kit (Becton Dickinson). The data were processed using ModFit LT program. Results: Comparative analysis of the cell cycle kinetics showed an increase in the percentage of cells in G2+M phase by 2 times in diploid and by 2.1 times in aneuploid recurrent tumors in comparison with primary ones (1.8±0.5% vs. 0.9±0.1% for diploid tumors; 5.4±2.2% vs. 2.6±0.7% for aneuploid tumors). An increase in the percentage of aneuploid tumors was found in recurrent G2 and G3 tumors (from 50% in primary to 66.7% in recurrent G2 tumors and from 63.25% in primary to 85% in recurrent G3 tumors). Mean content of aneuploid cells in recurrent G2 tumors was 2.2 times higher (p≤0.05), while the differences in primary and recurrent G3 tumors were not significant. The percentage of aneuploid tumors depended on the disease stage and increased in stages IIb and III in recurrent tumors, compared to primary ones (from 37.5% to 71.4% in recurrent st. IIb; and from 65% in primary st. III to 72.7% in recurrences) (p≤0.05). Conclusions: DNA analysis by flow cytometry demonstrated a high biologic potential of both primary and recurrent tumors. Some values in the mitotic cycle in recurrent tumors were probably associated with adjuvant therapy, as well as influenced by the coefficient of two parameters – the percentage of cells in G2+M phase and the cell loss factor determining a high malignant potential of these tumors.

scholarly journals Extracellular Adenine Nucleotides and Adenosine Modulate the Growth and Survival of THP-1 Leukemia Cells

2020 ◽  
Vol 21 (12) ◽  
pp. 4425
Author(s):  
Kamila Puchałowicz ◽  
Maciej Tarnowski ◽  
Marta Tkacz ◽  
Dariusz Chlubek ◽  
Patrycja Kłos ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Cell Growth ◽  
Cytotoxic Effect ◽  
Purinergic Signaling ◽  
Adenine Nucleotides ◽  
Leukemic Cell ◽  
Surface Expression ◽  
Boyden Chamber ◽  
Growth And Survival

A new approach to improve the effectiveness of acute myeloid leukemia (AML) treatment is to use the properties of purinergic signaling molecules secreted into the bone marrow milieu in response to leukemic cell growth. Therefore, our study aimed to evaluate the effects of extracellular adenine nucleotides and adenosine on the growth and death parameters in the leukemic THP-1 cell line. Cells were exposed to ATP, ADP, AMP, adenosine and nonhydrolyzable analogues of ATP and ADP (ATPγS and ADPβS) in a 1–1000 μM broad concentration range. The basal mRNA expression of the P1 and P2 receptors was evaluated by real-time PCR. Changes in the processes of cell growth and death were assessed by flow cytometry analysis of proliferation, cell cycle and apoptosis. Chemotaxis toward stromal cell-derived factor-1 (SDF-1) was performed using the modified Boyden chamber assay, and chemokine receptor type 4 (CXCR4) surface expression was quantified by flow cytometry. We indicated several antileukemic actions. High micromolar concentrations (100–1000 μM) of extracellular adenine nucleotides and adenosine inhibit the growth of cells by arresting the cell cycle and/or inducing apoptosis. ATP is characterized by the highest potency and widest range of effects, and is responsible for the cell cycle arrest and the apoptosis induction. Compared to ATP, the effect of ADP is slightly weaker. Adenosine mostly has a cytotoxic effect, with the induction of apoptosis. The last studied nucleotide, AMP, demonstrated only a weak cytotoxic effect without affecting the cell cycle. In addition, cell migration towards SDF-1 was inhibited by low micromolar concentrations (10 μM). One of the reasons for this action of ATPγS and adenosine was a reduction in CXCR4 surface expression, but this only partially explains the mechanism of antimigratory action. In summary, extracellular adenine nucleotides and adenosine inhibit THP-1 cell growth, cause death of cells and modulate the functioning of the SDF-1/CXCR4 axis. Thus, they negatively affect the processes that are responsible for the progression of AML and the difficulties in AML treatment.

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