Circ_RPL23A acts as a miR-1233 sponge to suppress the progression of clear cell renal cell carcinoma by promoting ACAT2
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Abstract Background Clear cell renal cell carcinoma (CCRCC) is a prevalent urological carcinoma with high metastatic risk. Circular RNAs (circRNAs) have been identified as effective diagnostic and therapeutic biomarkers for CCRCC. This research aims to disclose the involvement of circRNA ribosomal protein L23a (circ_RPL23A) in CCRCC, and how it regulates carcinogenesis. Methods We performed quantitative real-time polymerase chain reaction (qRT-PCR) to examine circ_RPL23A, microRNA-1233 (miR-1233) and acetyl-coenzyme A acetyltransferase 2 (ACAT2). Cellular behavior detection included cell cycle, proliferation, apoptosis and metastasis, which were severally analyzed using propidium iodide (PI)-flow cytometry, 3-(4, 5-dimethylthiazol-2-y1)-2, 5-diphenyl tetrazolium bromide (MTT), Annexin V/PI-flow cytometry and transwell migration/invasion assays. ACAT2 and related proteins of cell cycle, epithelial-mesenchymal transition (EMT) were measured via western blot. Target relationship was analyzed via dual-luciferase reporter assay. A xenograft model was used to study circ_RPL23A action in mice. Results Both in CCRCC tissues and cells, circ_RPL23A had a down-regulated tendency. Explicitly, circ_RPL23A overexpression inhibited cell cycle, proliferation and metastasis but enhanced apoptosis of CCRCC cells, whereas these effects of circ_RPL23A were dependent on the suppression of its target miR-1233. Besides, miR-1233 could target ACAT2 and circ_RPL23A regulated ACAT2 by sponging miR-1233. Also importantly, miR-1233 was a pro-cancer gene in CCRCC via targeting ACAT2. CCRCC tumor growth was also decreased in vivo by circ_RPL23A through miR-1233/ACAT2 axis. Conclusion Altogether, the circ_RPL23A/miR-1233/ACAT2 axis in this report provides an in-depth cognition for CCRCC pathogenesis and circ_RPL23A may has pivotal value in diagnosing and treating CCRCC.Keywords:
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Objective To investigate the mechanism underlying the anticancer activity of astragalus on human laryngeal cancer.Methods Hep-2 cells were treated with different concentrations of astragalus for 24 h.MTT assay was used to evaluate cell proliferation.Flow cytometry with PI staining and fluorescent microscopy with hoechst 33258 staining were used to estimate cell cycle distribution and cell apoptosis.Results Astragalus inhibited cellular proliferation in a dose dependent manner(P0.05).Flow cytometry analysis showed that treatment with astragalus resulted in accumulation of cells at the G2/M phase of the cell cycle and cell apoptosis in a dose dependent manner(P0.05).Marked apoptotic changes were observed by 33258 staining.Conclusion Astragalus inhibited cell proliferation and induced apoptosis of Hep-2 cells by regulating cell cycle,forming G2/M phase inhibition.
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Objective To explore the effect of Tanshinone ⅡA on cell proliferation,apoptosis and cell cycle of fibroblasts derived from keloid.Methods Fibroblasts derived from keloid were cultured with different concentration of Tanshinone ⅡA(0 μg/mL,50 μg/mL,100 μg/mL and 200 μg/mL) in vitro.CCK-8 was used to detect cell proliferation,and flow cytometry was used to analyze cell apoptosis,cell cycle at the different time.Results Cell proliferation of fibroblasts derived from keloid was decreased,apoptosis at the early phase was increased,and cell cycle in G0/G1 phase was arrested by different concentration of Tanshinone ⅡA with different intervention time(P0.001),especially in 200 μg/mL group.Variance analysis showed that those differences have statistical significance.Conclusion Tanshinon ⅡA can suppress cell proliferation,induce cell apoptosis and arrest cell cycle of fibroblasts derived from keloid.The effect of Tanshinone ⅡA was affected by different concentration and different intervention time.
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To study the mechanism of the effects of microcystin-LR on hepatocytes proliferation,dedifferentiation,cell cycle and apoptosis. In this study,MTT test,image analysis and flow cytometry were applied to assess the cell proliferation,cell spreading,cell cycle and apoptosis of primary cultured hepatocytes exposed to 0,0.1,1.0 or 10.0 μg/L MC-LR. Cell proliferation and cell spreading were promoted in a time and dose dependent manner following exposure to microcystin-LR. The accelerate rates of cell proliferation in the three tested groups were 102.47%,107.88% and 111.52%,respectively,compared with that of control group. Cell spreading ahead of schedule was detected in the tested groups and the means of cell surface in the tested groups increased as well. Synchronously,typical apoptotic changes as well as synchronization at the G 2/M-phase were both detected in the tested groups,the apoptosis rates of control group and the tested groups were 6.47%,22.20%,37.73% and 49.7%,respectively. [Conclusion] The results suggested that microcystin could affect hepatocyte cycles,and induce cell dedifferentiation and proliferation as well as cycle block,which might lead to cell apoptosis.
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Objective To investigate the effects of a novel ruthenium(Ⅱ) polypyridyl complex △-[Ru (phen)2MCMIP]2 + (△-1) on proliferation and apoptosis of human osteosarcoma cell line MG-63 in vitro.Methods Cell counting of kit-8 (CCK-8) assay was used to detect the proliferation of MG-63 cells after 24,48,72h treatment with △-1 under the concentrations of 0.00,12.50,25.00,50.00,100.00,150.00 μmol/L;Changes of apoptosis and cell cycle in MG-63 cells after 24 h treatment of 0.00,25.00,50.00,100.00 μmol/L A-1 were determined and analyzed by flow cytometry (FCM).Results Ruthenium (Ⅱ) polypyridyl complex A-1 could significantly inhibit the growth of MG-63 in a dose and time dependent manner; the IC50 of 24 h,48 h,72 h was 57.80μmol/L,45.27μmol/L,32.51μmol/L respectively; flow cytometry detection showed that A-1 induced 37.10％ of apoptosis,while only 1.06％ in control group,and arrested cell cycle at G0/G1 phase.Conclusion Ruthenium(Ⅱ) polypyridyl complex A-1 is able to inhibit proliferation and induce apoptosis in MG-63 cells and arrest cell cycle at G0/G1 phase.
Key words:
Ruthenium(Ⅱ) complex; MG-63 cells; Proliferation; Apoptosis; Cell cycle
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Objective
To investigate the effect of microRNA(miR)-487b on the proliferation and apoptosis of glioma cells by observing the changes of cell proliferation, cell cycle and apoptosis.
Methods
The expression of miR-487b in 6 samples of glioma tissues and 6 samples of non-tumor control brain tissues was detected by using real-time PCR (qRT-PCR). After artificially synthesized miR-487b mimics was transiently transfected into LN229 glioma cells, the expression of miRNA-487b was tested by qRT-PCR. MTT assay was applied to detect the cell proliferation. The cell cycle and apoptosis were measured by flow cytometry.
Results
MiR-487b was down-regulated in glioma samples(2.67±0.90)×10-2 compared with non-tumor samples(1.23±0.22). The transient transfection of miR-487b mimics into LN229 glioma cells significantly increased the expression of miR-487b (compared with normal control, P<0.05). The result of MTT assay showed that miR-487b might inhibit LN229 cells proliferation significantly (compared to normal control P<0.05). The cell cycle analysis detected by flow cytometry assay showed that miR-487b raised the cell proportion in G0/G1 phase (compared to normal control, P<0.05). The apoptosis rate detected by flow cytometry assay showed that miR-487b might promote the cell apoptosis (compared with normal control, P<0.05).
Conclusion
MiR-487b might decrease glioma cells proliferation, block cell cycle, and promote cell apoptosis, which indicated that miR-487b was a new target for the diagnosis and treatment of glioma.
Key words:
MicroRNA-487b; Glioma; Proliferation; Apoptosis
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Objective:To explore the effects of Ambrolic acid(AmbA) on proliferation and apoptosis of human gastric cancer SGC-7901 cell line.Methods:MTT assay was used to observe the anti-proliferation effect of AmbA on SGC-7901 cells with morphological changes observed under optical microscopy at the same time.The apoptosis rates and cell cycle status of SGC-7901 cells treated by AmbA were determined by flow cytometry(FCM).Results:AmbA inhibited the proliferation of SGC-7901 cells in a dose and time-dependent manner.FCM analysis showed that cells were arrested in S phase and induced apoptosis dose-dependently.Conclusion:AmbA has significant antitumor activity in vitro.The potential mechanism of anti-proliferation effect caused by AmbA may be cell-cycle arrest in S phase and cell apoptosis.
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Changes of apoptosis and cell cycle progression of HeLaS 3 cells following irradiation with different doses of X rays were observed. The percentages of apoptotic cells were examined by flow cytometry (FCM) after the cells had been stained with propidium iodine(PI) and Hoechst33342(HO), and the cell cycle was detected by FCM stained with only PI. It was found that the percentages of apoptotic cells and the cells in S phase and G 2/M phase increased after irradiation with 0.5-4.0Gy X rays. So irradiation may induce apoptosis and cell cycle block for HelaS 3 cell lines.
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We studied the effects of caffeine on cell viability, cell cycle profiles, proliferation, and apoptosis in rat C6 and human U87MG glioblastoma cell lines.Cell viability was quantified by the methyl thiazolyl tetrazolium (MTT) assay. Flow cytometry was used to quantify the relative number of cells in different phases of the cell cycle, while cell proliferation was quantified using the Cell Counting Kit-8. The proportion of apoptotic cells was determined by flow cytometry, and expression of apoptosis-related proteins Caspase-3, Cyt-C, Bax and Bcl-2 by Western blot.Caffeine at doses of up to 0.5 mM did not affect cell viability in both rat C6 and human U87MG glioblastoma cells. Further studies were done using the dose of 0.5 mM. Percentage of cells in the G0/G1 phase was markedly increased, while percentage of cells in the S phase decreased, after cell treatment with caffeine. Cell proliferation was significantly inhibited by caffeine. Furthermore, caffeine induced cell apoptosis, decreased expression of Bcl-2, and increased expression of Cyt-C and Caspase-3.Caffeine inhibits proliferation and induces apoptosis in glioblastoma cells. Our results provide the experimental basis for further studies of potential role of caffeine in the treatment of glioblastomas.
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Objective
To study the effect of miR-1908 on cell proliferation,cell cycle,apoptosis of human preadipocytes-visceral.
Methods
human preadipocytes-visceral were cultured and divided into three treatment groups—inhibitor group (transfected with chemically modified miR-1908-5p inhibitor), empty vector group (transfected empty carrier) and control group (without any treatment).CCK8 assay was used to detect the proliferation of HPA-v, Tunnel method was used to detect the apoptosis, Flow cytometry was used to detect cell cycle.
Results
CCK8 assay showed that the absorbance values of miR-1908 inhibitor group (0.61±0.07) was significantly lower than that of the control group (1.01±0.05) (P<0.05). Tunel results showed that the apoptotic rate of miR-1908 inhibitor group (0.27±0.03)% was significantly higher than that of control group (0.13±0.05)% (P<0.05). The results of flow cytometry showed that miR-1908 inhibitor might induce the G0/G1 phase cell cycle arrest.
Conclusions
Inhibition of miR-1908 expression could significantly inhibit HPA-v cell proliferation, induced apoptosis and change the cell cycle distribution.
Key words:
MiR-1908; Adipocytes; Proliferation; Apoptosis; Cell cycle
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To studay the effect of Inner Mongolia Drug Shengyangshiweiwan on cell apoptosis and cell cycle in humand gastric cancer MGC—803 cell,MTT assay was used to examine the proliferation inhibitory effect of shengyangshiyiweiwan on MGC—803 cell,and calculate the inhibitory rate of growth.Morphologic changes of apoptosis were observed by reverse discrepancy and offluorescent light microscope;flow cytometry was used to analysis cell cycle.MTT assay showed that shengyang has proliferation inhibiting effect on MGC—803 cell in dose and time dependent manners.The cells treated with shengyang(2.5 mg/mL) for 48 hours showed characters of apoptosis under light microscope.An 'sub-G1 peak was detected by flow cytometry,the result analysis of cell cycle indicated that the number of cells of S phase increased,that of G0/ G1 and G2/M phase decreased after treatment on MGC—803,it was arrested S phase.It is conclused that shengyang has proliferation inhibiting effect and may induce apoptosis in gastric cancer MGC—803 cell in a certain dose range,which may be one of the antitumor mechansim.
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