PMCID string | Title string | Sentences string |
|---|---|---|
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | CENP-B analysis showed that there were twice as many chromosomes present with 1µM alisertib in average, with most treated cells showing 46 chromosomal pairs, 4n-aneuploid, compared 23 chromosomal pairs 2n-normalploid with DMSO. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Following the same PI staining flow cytometry method used for identifying polyploidy, the effectiveness of Aurkin A to reduce alisertib induced polyploidy was evaluated. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | U2932 cells were treated with DMSO, 1µM alisertib, 100µM Aurkin A, or a combination of 1µM alisertib plus 100µM Aurkin A. Samples were taken on day three and five to track polyploidy development over time (Fig. 2a and b). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Flow cytometry showed a significant decrease of both 8n and 16n aneuploid cells populations with combination therapy compared to alisertib alone. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The reduction of polyploidy in the combination therapy was seen during both the three- and five-day time points. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Interestingly, no polyploidy was observed with Aurkin A or DMSO on day three and five. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Treatment with 1µM alisertib showed a large increase in 16n aneuploidy between days three and five (4% to 21%), and relatively similar levels of 8n aneuploidy (28% and 27%). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Combination treatment maintained a low level of 16n aneuploid cells at both time points (4% each), and increased levels of 8n aneuploid cells between days three and five (2% and 12%). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The 1mM alisertib groups showed a higher proportion of G2 to G1 cell population at day three (G1 16% and G2 25%) and day five (G1 14% and G2 21%). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The increased G2 peak is the result of 4n tetraploid cells accompanying the normal 4n G2 cells in the G2 peak. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | For combination therapy on day three, a larger proportion of the population is in G1 (43%) than G2 (38%). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | However, the DMSO control population maintains a high G1 population at 81%, and a lower G2 population at 14%. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | On day five, combination therapy group showed a G1 to G2 proportion similar to the alisertib treated group. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Fig. 2Disruption of Alisertib induced polyploidy by Aurkin A. U2932 cells were treated for three or five days with DMSO, Alisertib, Aurkin A, or a combination Alisertib and Aurkin A. Cells were harvested, stained with a propidium iodide solution, and analyzed via flow cytometry (A, B). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | To assess cellular apoptosis, a PI and FITC Annexin V apoptosis kit was utilized. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The FITIC Annex V conjugate will bind to phosphatidylserine present on the plasma membrane of early apoptotic cells (Annexin V positive/PI negative). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Necrotic cells with increased cell membrane compromise, will allow PI to enter the cell (Annexin V negative/PI positive). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | At later stages of apoptosis, phosphatidylserine will be present, along with a high degree of cell membrane comprise, enabling the binding of both Annexin V and PI (double positive, C). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Cells that do not uptake the Annex V or PI are considered viable cells. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Apoptosis analysis was performed on U2932 cells treated with DMSO, Alisertib, Aurkin A, or a combination of Alisertib and Aurkin A for five days (D). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | To determine cell viability, MTS assays were performed on U2932 (E), VAL, MDA-MB-231, Hela, Mia PaCa-2, U-2 OS, and Granta-519 cells, treated with either DMSO, Alisertib, Aurkin A, or a combination Alisertib and Aurkin A for four days. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Assays were performed in replicates of four and normalized to the DMSO treated condition. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Using the Loewe additivity method, the combination index was calculated to determine if the addition of 100µM Aurkin A was synergistic at each inhibitory concentration (F).Fig 2 Disruption of Alisertib induced polyploidy by Aurkin A. U2932 cells were treated for three or five days with DMSO, Alisertib, Aurkin A, or a combination Alisertib and Aurkin A. Cells were harvested, stained with a propidium iodide solution, and analyzed via flow cytometry (A, B). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | To assess cellular apoptosis, a PI and FITC Annexin V apoptosis kit was utilized. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The FITIC Annex V conjugate will bind to phosphatidylserine present on the plasma membrane of early apoptotic cells (Annexin V positive/PI negative). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Necrotic cells with increased cell membrane compromise, will allow PI to enter the cell (Annexin V negative/PI positive). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | At later stages of apoptosis, phosphatidylserine will be present, along with a high degree of cell membrane comprise, enabling the binding of both Annexin V and PI (double positive, C). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Cells that do not uptake the Annex V or PI are considered viable cells. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Apoptosis analysis was performed on U2932 cells treated with DMSO, Alisertib, Aurkin A, or a combination of Alisertib and Aurkin A for five days (D). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | To determine cell viability, MTS assays were performed on U2932 (E), VAL, MDA-MB-231, Hela, Mia PaCa-2, U-2 OS, and Granta-519 cells, treated with either DMSO, Alisertib, Aurkin A, or a combination Alisertib and Aurkin A for four days. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Assays were performed in replicates of four and normalized to the DMSO treated condition. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Using the Loewe additivity method, the combination index was calculated to determine if the addition of 100µM Aurkin A was synergistic at each inhibitory concentration (F). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | PI flow cytometry in VAL cells treated for five days with alisertib, Aurkin A, the combination or DMSO showed results comparable to the five-day U2932 treatment (Sup Fig. 2a). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | In VAL cells treated with DMSO vehicle, the G1 peak contained 56.5% of cells and G2 23.1%. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The 8n polyploid population was minimal (< 1%). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | 85µM Aurkin A treatment was similar to the control, with a G1 population of 60%, G2 27.5%, and 8n < 1%. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | VAL cells treated with 50nM alisertib had a G1 population of 27.4%, G2 39.6%, and 8n of 13.6%. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The combination alisertib and Aurkin A treatment showed a reduction in 8n polyploidy to 1.4% and a G1 and G2 population of 36% and 49.1% respectively, indicating that Aurkin A can revert or prevent alisertib-induced polyploidy. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | To assess whether the reduction observed in alisertib-induced polyploid cells is linked to increased apoptosis of the polyploid population, we performed an apoptosis analysis utilizing PI-annexin V stain and flow cytometry. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Samples untreated or treated with alisertib, Aurkin A, or combination concentrations were collected after five-days of treatment. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | In parallel, we assessed survival of treated cells after drug clearance. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | For this, cells were treated for 5 days, washed, and cultured an additional four days without drug. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | At day nine, all cells were harvested to assess post-treatment survival. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Both day five and nine samples were stained using a PI and FITC Annexin V apoptosis kit (Biolegend). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The FITC Annexin V conjugate binds to phosphatidylserine present on the plasma membrane of early apoptotic cells (Annexin V positive/PI negative). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Necrotic cells with increased cell membrane compromise allows PI to enter cells (Annexin V negative/PI positive). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | At latter stages of apoptosis, cells will have phosphatidylserine present along with a high degree of damage in the cell membrane enabling the binding of both Annexin V and PI (double positive). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Cells that do not uptake the Annexin V or PI are considered viable cells (double negative, Fig. 2c). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Five-day alisertib treated cells had a viable population of greater than 90% below 100nM, equivalent to the DMSO-treated control group (Fig. 2d). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The 500nM alisertib treated group saw a sharp decline in viability to 67%, along with the 1000nM and 5000nM at 54% and 39% respectively. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Interestingly, at day nine the 500nM population recovered having a cell viability of 82% (Sup Fig. 2b). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The 1000nM alisertib maintained a viability of 54% while the 5000nM group declined to a viable population of 22%. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Similar to the low dose alisertib groups, both the 10µM and 50µM Aurkin A treatments had a high cell viability comparable to the control. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The 100µM group had a lower cell viability of 72% and the 200µM Aurkin A group had a drastic decline to a viability of 14%. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The 10, 50, and 100µM populations on day nine all had similar viability to the DMSO control. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The 200µM Aurkin A group saw a slight improvement in cell viability to 21%. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The addition of 100µM Aurkin A to alisertib decreased cell viability in the lower dose alisertib connections. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | At day five, the 10nM and 100nM alisertib treatments had a viability of 95% and 92%, with the addition of Aurkin A, viability decreased to 67% and 59% respectively. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The largest viability change was seen in the 500nM group, which declined from 67% to 12% with the addition of Aurkin A. At day nine, both the 10nM and 100nM alisertib combination groups had recovered close to the level of the DMSO control. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | However, viability of the 500nM alisertib combination group remained low at 19%. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Notably, this decreased viability in the 500nM alisertib plus 100µM Aurkin A combination group was lower than both the 5000nM alisertib (22%) and 200uM Aurkin A (21%) at day nine demonstrating synergy between the two drugs. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | PI Annexin V staining on VAL cells treated with shows a similar reduction in cell viability in the combination treatment (Sup Fig. 2c). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Five-day DMSO treatment in VAL cells resulted in a 94% viable population. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Interestingly there was a decrease in viability in both the 50nM alisertib treatment and 85µM Aurkin A treatment to 48% and 55% viability respectively. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | However, the combination alisertib Aurkin A treatment declined to a viable population of 12.7%. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | To further investigate the cell killing potential of Aurkin A on DLBCL and other cell lines, MTS cell proliferation assays were performed. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | U2932, VAL, MDA-MB-231, Hela, Mia PaCa-2, U-2 OS, Granta-519 cells were treated with increasing concentrations of alisertib or Aurkin A for four days. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Cells were then assessed for cell proliferation via CellTiter 96 AQueous One Solution Cell Proliferation Assay. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | All experiments were performed in four replicates and normalized to DMSO control group. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | In U2932, the assay showed an IC50 of alisertib of 983nM and Aurkin A of 112µM. Using the determined IC50 of Aurkin A, a combination group treated with 100µM Aurkin A and increasing concentrations of alisertib was conducted (Fig. 2e and Sup Fig. 2d). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The combination group showed a significant reduction in IC50 of alisertib to 34nM (Fig. 2e). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Val cells showed a similar Aurkin A IC50 of 110µM, and a reduction in alisertib IC50 from 60nM to 2.64nM with the addition of 100µM Aurkin A (Sup Fig. 2e and f). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | MDA-MB-231, Hela, Mia PaCa-2, U-2 OS, Granta-519 cell lines also saw a significant drop in IC50 with the addition of 100µM Aurkin A (Table 1, Sup Fig. 2g–n).Table 1Summary of cell viability testing with combination alisertib and Aurkin A therapy in multiple cell lines. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | To determine effect of the combination therapy on different other cell types, MTS assays were performed on U2932, VAL, MDA-MB-231, Hela, Mia PaCa-2, U-2 OS, and Granta-519 cells treated with DMSO, Alisertib, Aurkin A, or a combination Alisertib and Aurkin A for four days. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Assays were performed in replicates of four and normalized to the DMSO treated condition. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Table 1Cell lineCancer typeAlisertib IC50 (nM)Aurkin A IC50 (µM)Alisertib IC50 with 100µM Aurkin A (nM)U2932Diffuse Large B-Cell Lymphoma (double expressor)98311233.7VALDiffuse Large B-Cell Lymphoma (double hit)601102.64MDA-MB-231Triple negative breast adenocarcinoma1186120160HelaCervical carcinoma110515184Mia PaCa-2Pancreatic carcinoma56610948U-2 OSOsteosarcoma976135108Granta-519Mantel Cell Lymphoma27512749 Summary of cell viability testing with combination alisertib and Aurkin A therapy in multiple cell lines. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | To determine effect of the combination therapy on different other cell types, MTS assays were performed on U2932, VAL, MDA-MB-231, Hela, Mia PaCa-2, U-2 OS, and Granta-519 cells treated with DMSO, Alisertib, Aurkin A, or a combination Alisertib and Aurkin A for four days. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Assays were performed in replicates of four and normalized to the DMSO treated condition. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | To evaluate alisertib and Aurkin A drug synergy in each cell line, the IC50, IC75, and IC90 of alisertib with and without 100µM Aurkin A was evaluated using the Loewe additivity method. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The calculated combination indexes were then plotted on an isobologram to show any potential synergy (Fig. 2f). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Combination indexes less than one are considered synergistic and indexes over one are antagonistic, with indexes further from one showing higher degrees of synergy/antagonism. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | All cell lines were found to have synergy at the IC50, IC75, and IC90 of alisertib with 100µM Aurkin A. Increased levels of synergy were seen at the IC75, and IC90, showing that significantly less alisertib is required to inhibit larger proportions of the cell population when Aurkin A is added. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Resistance to apoptosis, uncontrolled proliferation, resistance to cell cycle checkpoint activation, or the ability to reenter the cell cycle after extended periods in G0 are all known hallmarks of cancer. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Importantly, over-expression of cell cycle effector kinases can cause any or all these phenotypes, and thus are important contributing factors to carcinogenesis. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | We have established FUCCI cells to monitor cell cycle progression in VAL and U2932 cells over time, using live imaging. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Fluorescent markers were introduced with lentiviruses to generate stable cell lines expressing Geminin-clover (S/G2/M) and cdt1-mKO (G1), which can be monitored overtime in the green and red channels respectively of an Incucyte S3 (Essen) imaging system. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | First, the normal cell cycle progression of FUCCI U2932 cells was established (Fig. 3a). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Before the start of all experiments, FUCCI U2932 cells were treated with a double thymidine block to synchronize them at the end of the G1 phase. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | To establish normal cell cycle progression, FUCCI U2932 cells treated with DMSO were imaged every 30 minutes for five days. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The DMSO control is a representation of a normal cell cycle showing regular mitotic division approximately every 22 to 24 hours (representative images shown in Fig. 3b and Sup Fig. 3). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Due to the double thymidine synchronization, time point zero initially shows a red fluorescent cell, which progresses to yellow and green within four hours of monitoring (transition from G1-S-G2). |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The cell continues to grow and divides into two cells after 16 hours and enters G1. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The daughter cells progress through a normal cell cycle and undergo another round of mitotic division at 37 and 41 hours. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | The cells progress to G2 phase but fail to divide a third time and begin apoptotic blebbing at 70 hours. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Fig. 3Analysis cell cycle progression using FUCCI U2932 cells. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | A depicts normal cell cycle progression of FUCCI cells, along with corresponding images of a DMSO control cell. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | During G1, red fluorescent cdt1-mKO is expressed which transitions to green as Geminin-clover is expressed during S-G2. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | U2932 cells were treated synchronized with a double thymidine block over a two-day period. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | Cells were then released from the block by washing with fresh media, supplemented with 1 % final volume DMSO (B).Fig 3 Analysis cell cycle progression using FUCCI U2932 cells. |
PMC11225860 | Aurkin-A, a TPX2-aurora a small molecule inhibitor disrupts Alisertib-induced polyploidy in aggressive diffuse large B cell lymphoma | A depicts normal cell cycle progression of FUCCI cells, along with corresponding images of a DMSO control cell. |
Subsets and Splits
No community queries yet
The top public SQL queries from the community will appear here once available.