2015•Zhonghua shenjing waike zazhiRequires access

The repressive effect of antisense miR-106b on human glioma cell growth and invasive ability

Minhua Ye, Anling Zhang, Lei Wu, Kun Wang, Guangxiu Wang, Zhifan Jia, Qiang Huang

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Abstract

Objective To study the effect of miR-106b inhibitor on glioma cell line growth and invasive ability in vitro. Methods The miR-106b inhibitor was transfected by liposome to repress the expression of miR-106b. qRT-PCR was taken to measure miR-106b expression after tansfection. The cell cycle kinetics and cell proliferation rate were detected by flowcytometry and MTT assay, the cell malignant proliferative ability was evaluated by soft agar assay, and the invasive ability was detected by transwell assay. Results The antisense oligonucleotide of miR-106b effectively down-regulated miR-106b expression in glioma cells. The progression of cell cycle was blocked. The rate of S phase decreased, and the rate of G0/G1 phase increased. The distribution of cell cycle was significantly different in U251 and TJ905 (P<0.05), but not in LN229. After transfection for 48h, the proliferation activity of three cell lines was obviously inhibited. Also, the cell clone formation was repressed[The rate of clone efficiency: U251: (10.6 ± 0.9)%、(10.5 ± 1.2)% vs.(3.3 ± 0.4)%; LN229: (12.5 ± 1.7)%、(11.0 ± 2.4)% vs.(4.1 ± 0.3)%; TJ905: (9.9 ± 1.2)%、(10.2 ± 0.6)% vs.(3.5 ± 0.4)%. P< 0.01]. The invasive ability was also significantly repressed in cells transfected with miR-106b inhibitor as compared to those of the cells transfected with negative control oligonucleotide and control cells[The cell number per view was U251: (90.6±5.7), (85.2±6.1) vs .(27.6±4.0); LN229: (22.6±3.7), (21.2±3.7) vs. (2.4±1.5); TJ905: (79.4±4.4), (76.8±5.9) vs.( 11.8±3.2). P<0.01]. Conclusions Suppression of miR-106b expression could inhibit the proliferative and invasive ability of glioma cells. Key words: MicroRNAs; RNA, antisense; Glioma; Cell proliferation; Neoplasm invasiveness; In vitro

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Objective To study the effect of miR-106b inhibitor on glioma cell line growth and invasive ability in vitro. Methods The miR-106b inhibitor was transfected by liposome to repress the expression of miR-106b. qRT-PCR was taken to measure miR-106b expression after tansfection. The cell cycle kinetics and cell proliferation rate were detected by flowcytometry and MTT assay, the cell malignant proliferative ability was evaluated by soft agar assay, and the invasive ability was detected by transwell assay. Results The antisense oligonucleotide of miR-106b effectively down-regulated miR-106b expression in glioma cells. The progression of cell cycle was blocked. The rate of S phase decreased, and the rate of G0/G1 phase increased. The distribution of cell cycle was significantly different in U251 and TJ905 (P<0.05), but not in LN229. After transfection for 48h, the proliferation activity of three cell lines was obviously inhibited. Also, the cell clone formation was repressed[The rate of clone efficiency: U251: (10.6 ± 0.9)%、(10.5 ± 1.2)% vs.(3.3 ± 0.4)%; LN229: (12.5 ± 1.7)%、(11.0 ± 2.4)% vs.(4.1 ± 0.3)%; TJ905: (9.9 ± 1.2)%、(10.2 ± 0.6)% vs.(3.5 ± 0.4)%. P< 0.01]. The invasive ability was also significantly repressed in cells transfected with miR-106b inhibitor as compared to those of the cells transfected with negative control oligonucleotide and control cells[The cell number per view was U251: (90.6±5.7), (85.2±6.1) vs .(27.6±4.0); LN229: (22.6±3.7), (21.2±3.7) vs. (2.4±1.5); TJ905: (79.4±4.4), (76.8±5.9) vs.( 11.8±3.2). P<0.01]. Conclusions Suppression of miR-106b expression could inhibit the proliferative and invasive ability of glioma cells. Key words: MicroRNAs; RNA, antisense; Glioma; Cell proliferation; Neoplasm invasiveness; In vitro

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Available abstract

Objective To study the effect of miR-106b inhibitor on glioma cell line growth and invasive ability in vitro. Methods The miR-106b inhibitor was transfected by liposome to repress the expression of miR-106b. qRT-PCR was taken to measure miR-106b expression after tansfection. The cell cycle kinetics and cell proliferation rate were detected by flowcytometry and MTT assay, the cell malignant proliferative ability was evaluated by soft agar assay, and the invasive ability was detected by transwell assay. Results The antisense oligonucleotide of miR-106b effectively down-regulated miR-106b expression in glioma cells. The progression of cell cycle was blocked. The rate of S phase decreased, and the rate of G0/G1 phase increased. The distribution of cell cycle was significantly different in U251 and TJ905 (P<0.05), but not in LN229. After transfection for 48h, the proliferation activity of three cell lines was obviously inhibited. Also, the cell clone formation was repressed[The rate of clone efficiency: U251: (10.6 ± 0.9)%、(10.5 ± 1.2)% vs.(3.3 ± 0.4)%; LN229: (12.5 ± 1.7)%、(11.0 ± 2.4)% vs.(4.1 ± 0.3)%; TJ905: (9.9 ± 1.2)%、(10.2 ± 0.6)% vs.(3.5 ± 0.4)%. P< 0.01]. The invasive ability was also significantly repressed in cells transfected with miR-106b inhibitor as compared to those of the cells transfected with negative control oligonucleotide and control cells[The cell number per view was U251: (90.6±5.7), (85.2±6.1) vs .(27.6±4.0); LN229: (22.6±3.7), (21.2±3.7) vs. (2.4±1.5); TJ905: (79.4±4.4), (76.8±5.9) vs.( 11.8±3.2). P<0.01]. Conclusions Suppression of miR-106b expression could inhibit the proliferative and invasive ability of glioma cells. Key words: MicroRNAs; RNA, antisense; Glioma; Cell proliferation; Neoplasm invasiveness; In vitro

Key concepts: Transfection, Cell cycle, Cell growth, Molecular biology, Glioma, clone (Java method), Cell culture, Cell

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