Anti-proliferation and apoptosis-inducing effects of sodium aescinate on retinoblastoma Y79 cells

AIM: To explore the effect of the Andrographis paniculata (A. paniculata) polysaccharide on the proliferation and apoptosis of human retinoblastoma (RB) Y79 cells and its mechanism. METHODS: The refined A. paniculata polysaccharide was obtained using techniques such as water extraction, ethanol precipitation, and decompression concentration. The inhibition effect of the A. paniculata polysaccharide on the proliferation of Y79 cells was detected by cell proliferation assay. Flow cytometry was used for the detection of cell apoptosis rate and cycle change. Real-time qunatitative polymerase chain reaction (RT qPCR)and Western blotting were used to detect the expression of cell apoptosis signal pathway-related factors (caspase-3, caspase-8, and caspase-9) and cell cycle signal pathway-related factors (CDK1 and cyclinB1) at the transcriptional and translational levels. RESULTS: Infrared and ultraviolet spectrum scanning showed that the extracted drug was a polysaccharide with high purity. After being treated with different concentrations of A. paniculata polysaccharide for different periods of time, the Y79 cells showed different degrees of proliferation inhibition. Flow cytometric observations showed that the cell apoptosis rate and the proportion of cells blocked in the G2/M phase were significantly increased after A. paniculata polysaccharide treatment. Further analysis revealed that the mRNA and protein expression of caspase-3, caspase-8, and caspase-9 in the A. paniculata polysaccharide treatment groups increased significantly compared with that in the control groups, while the expression of CDK1 and cyclinB1 decreased significantly. CONCLUSION: The A. paniculata polysaccharide could inhibit the proliferation and induce apoptosis of Y79 cells. Its possible mechanism is via the upregulation of caspase-3, caspase-8, and caspase-9 expression in the cell apoptotic signaling pathway and the downregulation of CDK1 and cyclinB1 expression in the cell cycle signaling pathway.

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Knockdown of lncRNA HOTTIP Inhibits Retinoblastoma Progression by Modulating the miR-101-3p/STC1 Axis

Technology in Cancer Research & Treatment ◽

10.1177/1533033821997831 ◽

2021 ◽

Vol 20 ◽

pp. 153303382199783

Author(s):

XiangWen Yuan ◽

Zhaoyan Sun ◽

Congxian Cui

Keyword(s):

Cell Proliferation ◽

Western Blotting ◽

Underlying Mechanism ◽

Luciferase Reporter ◽

Luciferase Reporter Gene ◽

Qrt Pcr ◽

Gene Assay ◽

Proliferation And Apoptosis ◽

Y79 Cells

Objective: Retinoblastoma (RB) is a frequent eye cancer in children. Long non-coding RNA (LncRNA) HOXA transcript at the distal tip (HOTTIP) is aberrantly expressed in cancer tissues. This study explores the underlying mechanism of lncRNA HOTTIP in RB. Methods: HOTTIP expression in normal retinal cells and RB cell lines was detected using qRT-PCR. The proliferation of RB cells was measured using CCK-8 and EdU assays, and apoptosis was detected using flow cytometry and Western blotting after the transfection of si-HOTTIP into Y79 cells and pc-HOTTIP into HXO-RB-44 cells. The target relationships between HOTTIP and miR-101-3p, and miR-101-3p and STC1 were predicted by bioinformatics website and verified using dual-luciferase reporter gene assay. The binding of HOTTIP and miR-101-3p was verified using RNA pull-down assay. STC1 mRNA and protein in RB cells were measured using qRT-PCR and Western blotting. Moreover, si-HOTTIP and in-miR-101-3p/in-NC, and si-HOTTIP and pc-STC1/pcDNA were co-transfected into Y79 cells respectively to evaluate cell proliferation and apoptosis. Xenograft study was conducted, and Ki67-positive expression was detected using immunohistochemical staining. Results: HOTTIP expression was promoted in RB tissues and cells. Downregulation of HOTTIP inhibited proliferation and promoted apoptosis of Y79 cells, while upregulation of HOTTIP promoted proliferation and inhibited apoptosis of HXO-RB-44 cells. There were target relationships between HOTTIP and miR-101-3p, and miR-101-3p and STC1. Inhibition of miR-101-3p or overexpression of STC1 reversed the effect of si-HOTTIP on the proliferation and apoptosis of RB cells. Xenograft study showed that knockdown of HOTTIP suppressed the growth of RB in vitro. Conclusion: It could be concluded that HOTTIP sponged miR-101-3p to upregulate STC1 expression, thereby promoting RB cell proliferation and inhibiting apoptosis.

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Knockdown of LncRNA SNHG14 regulates proliferation and apoptosis via activating miR-204-5p/HMGA1 axis in retinoblastoma

10.21203/rs.2.24066/v1 ◽

2020 ◽

Author(s):

Xuedong Chen ◽

Xianling Tang ◽

Shiyong Zhao ◽

Yongbin Yu ◽

Qingjun Li ◽

...

Keyword(s):

Cell Proliferation ◽

Treatment Strategies ◽

Xenograft Model ◽

High Mobility ◽

Luciferase Reporter ◽

Reaction Cell ◽

High Mobility Group Protein ◽

Hmga1 Expression ◽

Proliferation And Apoptosis ◽

Y79 Cells

Abstract Background: Retinoblastoma (RB) is an aggressive intraocular malignancy of infant and childhood, which seriously endangers the vision and life of children. Long non-coding RNA Small Nucleolar RNA Host Gene 14 (lncRNA SNHG14) as a novel oncogene is involved in the control of cancer cell progression. However, the effects and molecular mechanism of SNHG14 on retinoblastoma remain confusing.Methods: Levels of SNHG14, high mobility group protein A1 (HMGA1) mRNA and microRNA (miR)-204-5p were detected by quantitative real-time polymerase chain reaction. Cell proliferation and apoptosis were measured by CKK-8 assay or flow cytometry, respectively. Western blot was used to detect HMGA1 and apoptosis-related protein. The interaction between miR-204-5p and SNHG14 or HMGA1 was explored by luciferase reporter assay. Murine xenograft model was established using Y79 cells stably transfected with sh-SNHG14.Results: SNHG14 and HMGA1 were up-regulated in retinoblastoma tissues and cell lines, and knockdown of SNHG14 or HMGA1 suppressed cell proliferation and induced cell apoptosis in retinoblastoma. MiR-204-5p was confirmed to directly bind to SNHG14 and HMGA1, SNHG14 positively regulated HMGA1 expression via miR-204-5p. Importantly, the anti-tumor effects mediated by SNHG14 knockdown could be reversed by miR-204-5p inhibition or HMGA1 overexpression in retinoblastoma cells. Furthermore, xenograft model showed SNHG14 silence impeded tumor growth in vivo.Conclusion: Knockdown of SNHG14 suppressed retinoblastoma progression by regulating miR-204-5p/HMGA1 axis, revealing a potential target to develop appropriate treatment strategies for retinoblastoma.

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Effect of Bruceine D on MicroRNA-155 Expression and Proliferation and Apoptosis of Human Retinoblastoma Cells

Nanoscience and Nanotechnology Letters ◽

10.1166/nnl.2020.3094 ◽

2020 ◽

Vol 12 (2) ◽

pp. 178-183

Author(s):

Dongju Qin ◽

Xiamuxiya Ainiwaer ◽

Huling Pan ◽

Qian Sha ◽

Xiaojuan Zhou

Keyword(s):

Caspase 3 ◽

Control Group ◽

Blank Control Group ◽

Cell Hyperplasia ◽

Apoptosis Rate ◽

Retinoblastoma Cells ◽

Proliferation And Apoptosis ◽

Human Retinoblastoma ◽

Y79 Cells

This study aimed to investigate the effect of bruceine D on the proliferation and apoptosis of human retinoblastoma cells and its effect on miR-155 expression. Y79 human retinoblastoma cells were cultured in vitro and randomly divided into the blank control group, DMSO (Dimethyl Sulphoxide) group, bruceine D 10 μmol/L group, bruceine D 20 μmol/L group, and bruceine D 40 μmol/L group. MTT assay was used to detect cell hyperplasia, flow cytometry to detect the apoptosis rate, and qRT-PCR to detect the expression level of miR-155. Anti-miR-155 or miR-155 mimics were transfected into Y79 cells, and hyperplasia and apoptosis rates were detected by the above methods. Western blotting was used to detect Bcl-2, Bax, and caspase-3 expression. Bruceine D can reduce the viability of Y79 cells (P < 0.05), significantly increase the apoptosis rate (P < 0.05), promote the expression of Bax and caspase-3 (P < 0.05), and inhibit miR-155 and Bcl-2 (P < 0.05). Compared with the anti-miR-con group, in the anti-miR-155 group, cell vigor was evidently reduced (P < 0.05), the apoptosis rate was evidently increased (P < 0.05), and the Bcl-2 level was reduced (P < 0.05). Bax and caspase-3 levels were evidently increased (P < 0.05). Transfection with miR-155 mimics can reduce the influence of caspase D-3 in hyperplasia and apoptosis of Y79 cells. Bruceine D may reduce the hyperplasia of human retinoblastoma cells and regulate cell apoptosis by downregulating miR-155 expression.

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