To investigate the expression and clinical significance of microRNA-218(miR-218)in human cervical cancer and the effects ofmiR-218 on proliferation, cell apoptosis and invasion of HeLa cells.QRT-PCR was used to detect the expression ofmiR-218 in 23 cases of normal cervical tissues and 114 cases of cervical cancer, and the relationship between the expression and the clinicopathological features was analyzed; HeLa cells were devided into three groups: non transfection (control group), transfected with empty liposomes negative control goup (NC group), transfected with miR-218 mimic (miR-218M group). The cell growth inhibiting ratio of HeLa cells was assessed by MTT assay. Fluorescence activated cell sorting was used to measure cell apoptosis. Changes of cell migration ability were detected by wound healing test and Transwell assay. QRT-PCR and Western blot were used to detect the expression of Bcl-2, Bax, NF-κB and E-cadherin, respectively.The expression ofmiR-218 in cervical cancer was down regulated, and there were significant differences in the different pathological types, stages, lymph node metastasis and interstitial infiltration in cervical cancer tissues ( P<0.01); After being transfected with miR-218 mimic, the proliferation of HeLa cells was significantly inhibited. The ability of invasion was decreased. QRT-PCR and Western blot showed that after being transfected with miR-218 mimic, the expression levels of Bcl-2 mRNA and protein were down-regulated and Bax mRNA and protein expression levels were increased, E-cadherin mRNA and protein expression were up-regulated, but NF -κB mRNA and protein expression were down-regulated.he low-expression of miR-218 is correlated with the poor clinicopathological features in human cervical cancer. MiR-218 overexpression reduces cancer cell proliferation and induces apoptosis and inhibits cell migration, suggesting that miR-218 may play a key role in the progression of human cervical cancer.
Abstract: Gastric cancer is the second leading cause of cancer-related death worldwide, with a poor response to current chemotherapy. Danusertib is a pan-inhibitor of the Aurora kinases and a third-generation Bcr-Abl tyrosine kinase inhibitor with potent anticancer effects, but its antitumor effect and underlying mechanisms in the treatment of human gastric cancer are unknown. This study aimed to investigate the effects of danusertib on cell growth, apoptosis, autophagy, and epithelial to mesenchymal transition and the molecular mechanisms involved in human gastric cancer AGS and NCI-N78 cells. The results showed that danusertib had potent growth-inhibitory, apoptosis-inducing, and autophagy-inducing effects on AGS and NCI-N78 cells. Danusertib arrested AGS and NCI-N78 cells in G 2 /M phase, with downregulation of expression of cyclin B1 and cyclin-dependent kinase 1 and upregulation of expression of p21 Waf1/Cip1, p27 Kip1, and p53. Danusertib induced mitochondria-mediated apoptosis, with an increase in expression of proapoptotic protein and a decrease in antiapoptotic proteins in both cell lines. Danusertib induced release of cytochrome c from the mitochondria to the cytosol and triggered activation of caspase 9 and caspase 3 in AGS and NCI-N78 cells. Further, danusertib induced autophagy, with an increase in expression of beclin 1 and conversion of microtubule-associated protein 1A/1B-light chain 3 (LC3-I) to LC3-II in both cell lines. Inhibition of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) and p38 mitogen-activated protein kinase pathways as well as activation of 5' AMP-activated protein kinase contributed to the proautophagic effect of danusertib in AGS and NCI-N78 cells. SB202191 and wortmannin enhanced the autophagy-inducing effect of danusertib in AGS and NCI-N78 cells. In addition, danusertib inhibited epithelial to mesenchymal transition with an increase in expression of E-cadherin and a decrease in expression of N-cadherin in both cell lines. Taken together, danusertib has potent inducing effects on cell cycle arrest, apoptosis, and autophagy, but has an inhibitory effect on epithelial to mesenchymal transition, with involvement of signaling pathways mediated by PI3K/Akt/mTOR, p38 mitogen-activated protein kinase, and 5' AMP-activated protein kinase in AGS and NCI-N78 cells. Keywords: danusertib, gastric cancer, Aurora kinase, apoptosis, autophagy, epithelial to mesenchymal transition
To observe the effect of autophagy on paclitaxel-induced CaSki cell death through the regulation of the expression of autophagy gene Beclin1, and to explore the interaction and relationship between autophagy and apoptosis.Eukaryotic expression vector pcDNA3.1-Beclin1 and RNA interference vector pSUPER-Beclin1 were transfected into human cervical cancer CaSki cells in vitro and screened for stable expression cell lines. The formation of autophagic vacuoles was observed with an electronic microscope. The expression of Beclin1 and LC3 was measured by Western blot. After being treated with paclitaxel, the change of cell proliferation was assessed by MTT assay, the percentage of apoptotic cells and autophagic cells were analyzed by flow cytometry.A lot of autophagic vacuoles were observed in pcDNA3.1-Beclin1 cells by electronic microscopy. Beclin1 and LC3 protein expression was up-regulated in CaSki cells transfected with pcDNA3.1-Beclin1, and was inhibited in cells transfected with pSUPER-Beclin1. MTT assay revealed the survival rate of CaSki cells was significantly decreased after being transfected with pcDNA3.1-Beclin1. After being treated with paclitaxel, the percentages of apoptotic cells and autophagic cells were both increased in pcDNA3.1-Beclin1 group compared with that of the blank control group especially the increase of apoptosis was particularly evident.Autophagy and apoptosis have different roles in the process of paclitaxel-induced cervical cancer CaSki cell line death. Overexpression of Beclin1 in CaSki cells may enhance the apoptosis induced by paclitaxel.
Glioblastoma (GBM) is the most common brain tumor with poor response to current therapeutics. Alisertib (ALS), a second-generation selective Aurora kinase A (AURKA) inhibitor, has shown potent anticancer effects on solid tumors in animal studies. This study aimed to investigate the killing effect of ALS on GBM cell line DAOY and the possible underlying mechanisms using both bioinformatic and cell-based approaches. Our molecular docking showed that ALS preferentially bound AURKA over AURKB via hydrogen bond formation, charge interaction, and π-π stacking. ALS also bound key regulating proteins of cell cycle, apoptosis and autophagy, such as cyclin-dependent kinase 1 (CDK1/CDC2), CDK2, cyclin B1, p27 Kip1, p53, cytochrome C, cleaved caspase 3, Bax, Bcl-2, Bcl-xl, phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), mammalian target of rapamycin (mTOR), 5'-adenosine monophosphate-activated protein kinase (AMPK), p38 mitogen-activated protein kinase (MAPK), beclin 1, phosphatase and tensin homolog (PTEN), and microtubule-associated protein light chain 3 (LC3). ALS exhibited potent growth-inhibitory, pro-apoptotic, and pro-autophagic effects on DAOY cells in a concentration-dependent manner. Notably, ALS remarkably induced G2/M arrest mainlyvia regulating the expression of CDK1/CDC2, CDK2, cyclin B1, p27 Kip1, and p53 in DAOY cells. ALS significantly induced the expression of mitochondria-mediated pro-apoptotic proteins such as Baxbut inhibited the expression of anti-apoptotic proteins such as Bcl-2 and Bcl-xl, with a significant increase in the release of cytochrome C and the activation of caspases 3 and 9. ALS also induced PI3K/Akt/mTOR and p38 MAPK signaling pathways while activating the AMPK signaling pathway. Taken together, these findings indicate that ALS exerts a potent inhibitory effect on cell proliferation and induces mitochondria-dependent apoptosis and autophagy with the involvement of PI3K/Akt/mTOR- and p38 MAPK-mediated signaling pathways in DAOY cells. ALS is a promising anticancer agent for GBM treatment.
Google Earth (GE) provides accurate and reliable global high-resolution images and can obtain the coordinates of any point on Earth's surface.A digital elevation model (DEM) is a dataset that quantitatively reflects the elevation of Earth's surface.Although GE and DEMs can be used to obtain the coordinates of any position on Earth, their open-access data can be affected by various factors, thereby inducing undesirable precision variability.Therefore, it is essential to estimate the accuracy of GE and open-source DEMs.In this study, 325 high-precision GPS survey points in 16 regions in China were used to evaluate the horizontal and vertical accuracies of GE and the elevation accuracy of two open-source DEMs.GE had a high horizontal accuracy with a root mean square error (RMSE) of 2.495 m and an error range of 1.090-4.844m.The elevation accuracy of GE (RMSE = 2.610 m) was lower than those of TanDEM-X (RMSE = 2.055 m) and AW3D30 (RMSE = 2.373 m) DEMs.At the same time, the impacts of slope, aspect, and feature type on the accuracy of these data are studied and analyzed.The results show that the accuracy of control data are closely related to the characteristics of the study area.Overall, these findings indicate that for future studies in China, GE can be used to acquire horizontal data, whereas TanDEM-X and AW3D30 are more suitable for elevation data that have higher precision and provide a reference for relevant research on geographic information.
Geometric calibration is an essential technique that improves the absolute positioning accuracy of synthetic aperture radar (SAR) imagery. Geometric calibration of SAR has recently been moving toward a field-free approach. Field-free geometric calibration leverages the consistency in positioning conjugate image points across multiple images to determine ground coordinates and geometric calibration parameters. Notably, when the number of available images is limited, field-free geometric calibration may present instability in determining calibration parameters. We conduct a detailed analysis of the various sources of error that can affect SAR geometric positioning. In addition, we introduce the concept of dilution of precision (DOP) to assess the accuracy of the obtained calibration parameters. When the DOP is less than 5, we believe the geometric calibration parameters to be accurate and reliable. We analyze the impact of satellite geometry distribution and calibrated image quantity on DOP. Out of the 18 YG-13 satellite images captured in the Songshan area of Henan province, 3 and 4 images were chosen for geometric calibration, resulting in 816 and 3060 potential combinations, respectively. Calibrated solutions are obtained for each combination, and combinations with inadequate assessment accuracy are filtered out based on the DOP. After getting the geometric calibration parameters from the solution, we compensate them in the positioning equation and compare the accuracy of the two checkpoints before and after screening. The findings demonstrate that the post-screening combination surpasses the pre-screening combination and achieves comparable performance to field-free calibration and cross calibration concerning image localization accuracy. The effectiveness of DOP in evaluating the accuracy of geometric calibration parameter estimation has been corroborated.
To investigate the role of Beclin 1 expression on the cisplatin-induced apoptosis in cervical cancer CaSki cells and to explore the potential mechanism underlying this effect.After overexpression or partial silencing of Beclin 1 in cervical cancer CaSki cells, the transfected group and the control group were treated with cisplatin for 24 hours. The percentage of apoptotic cells were assessed by flow cytometry. The mitochondrial membrane potential and activities of caspase-8/9/3 were detected by JC-1 fluorescence staining and colorimetry. The expression of cytochrome c was measured using a Western blot. The messenger RNA expression of Bax and Bcl-2 were detected by real-time quantitative reverse transcription polymerase chain reaction.Expression of Beclin 1 protein was up-regulated in overexpressed transfectants of CaSki cells. After treatment with cisplatin, the Beclin 1 overexpression group led to the decrease of mitochondrial membrane potential and increase of activities of caspase-9 and caspase-3, and showed a greater increase in apoptosis than did the nontransfected group. Furthermore, Beclin 1 overexpression resulted in increased cytoplasmic cytochrome c and Bax expression and decreased mitochondrial cytochrome c and Bcl-2 expression.Overexpression of Beclin 1 in CaSki cells may influence cisplatin-induced apoptosis by mitochondrial dependent pathway.
To investigate the effect of Beclin 1, an autophagy gene, on the expression of angiopoietin (Ang) protein and the Tie-2 receptor in CaSki human cervical cancer cells.Beclin 1 overexpression (pcDNA3.1-Beclin1) and knockdown (pSUPER-Beclin1) plasmids were independently transfected into CaSki cells, and stably transfected cells were selected. Real-time fluorescence quantitative PCR and Western blot analyses were performed to detect the mRNA and protein expression of Beclin 1, Ang-1, Ang-2, and Tie-2. MTT assays were employed to determine cell proliferation rates.In the cells transfected with pcDNA3.1-Beclin1, the expression of Beclin 1, Ang-2, and Tie-2 was markedly increased, but expression of Ang-1 was dramatically reduced. MTT assays revealed that the proliferation of these cells was also significantly suppressed. In the CaSki cells transfected with pSUPER-Beclin1, the expression of Beclin 1, Ang-2, and Tie-2 was inhibited.Overexpression of Beclin 1 can inhibit the proliferation of CaSki cells, which may be attributed to an imbalance among the expression of Ang-1, Ang-2 and Tie-2.
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