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400042 重庆, 陆军军医大学(第三军医大学)第三附属医院(野战外科研究所)肝胆外科
[陈平, 电话:(023)68757966, E-mail:chenpingsyd@126.com CHEN Ping,E-mail:chenpingsyd@126.com
收稿:2017-12-11,
修回:2018-01-04,
纸质出版:2018-05-15
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林夏, 朱元鑫, 付航玮, 等. Hsa_circ_0000711在人肝癌细胞系中的功能研究[J]. 第三军医大学学报, 2018,40(9):788-794.
Xia LIN, Yuanxin ZHU, Hangwei FU, et al. Function of circular RNA hsa_circ_0000711 in hepatocellular carcinoma cell lines[J]. Journal of Third Military Medical University, 2018, 40(9): 788-794.
林夏, 朱元鑫, 付航玮, 等. Hsa_circ_0000711在人肝癌细胞系中的功能研究[J]. 第三军医大学学报, 2018,40(9):788-794. DOI: 10.16016/j.1000-5404.201712097.
Xia LIN, Yuanxin ZHU, Hangwei FU, et al. Function of circular RNA hsa_circ_0000711 in hepatocellular carcinoma cell lines[J]. Journal of Third Military Medical University, 2018, 40(9): 788-794. DOI: 10.16016/j.1000-5404.201712097.
目的
2
研究环状RNA hsa_circ_0000711在人肝癌细胞系中的表达以及对肝癌细胞生物学功能的影响。
方法
2
利用RT-qPCR检测环状RNA hsa_circ_0000711在肝癌细胞系的表达; 通过小干扰RNA特异性敲低hsa_circ_0000711的表达; 通过CCK-8实验检测肝癌细胞系的增殖情况; 利用流式细胞技术检测细胞周期及细胞凋亡。Western blot检测下调hsa_circ_0000711后Cyclin D1、CDK4、cleavaed Caspase 3及Bcl-2蛋白的表达水平。
结果
2
Hsa_circ_0000711在肝癌细胞HepG2和SMMC-7721中明显高表达(
P
<
0.05)。与对照组相比
hsa_circ_0000711干扰组的肝癌细胞增殖能力显著降低(
P
<
0.05)
处于G
1
期细胞增多(
P
<
0.05)
且凋亡细胞显著增加(
P
<
0.05)。干扰hsa_circ_0000711的表达后
肝癌细胞中Cyclin D1、CDK4和Bcl-2的蛋白表达量显著降低(
P
<
0.05)
cleavaed Caspase 3蛋白表达量显著升高(
P
<
0.05)。
结论
2
Hsa_circ_0000711可能通过调控细胞周期及凋亡影响肝癌细胞的增殖。
Objective
2
To explore the expression profile of circular RNA (circRNA)
hsa_circ_0000711
in hepatocellular carcinoma (HCC) cell lines and determine the effect of hsa_circ_0000711 on the biological behaviors of HCC cells.
Methods
2
The expression level of Hsa_circ_0000711 were detected by real-time polymerase chain reaction assay (RT-qPCR) in HCC cell lines HepG2 and SMMC-7721 and human live cell line L02.Small RNA interference was used to knock down the expression of hsa_circ_0000711.Then CCK-8 assay was employed to detect the proliferation
and flow cytometry for cell cycle and apoptosis.The protein levels of Cyclin D1
CDK4
cleaved Caspase 3 and Bcl-2 were measured by Western blotting.
Results
2
Hsa_circ_0000711 was more strongly expressed in the HepG2 and SMMC-7721 cells than the L02 cells (
P
<
0.05).Down-regulation of hsa_circ_0000711 significantly suppressed the growth rate of HepG2 and SMMC-7721 cells (
P
<
0.05)
arrested the cells at cycle G
1
(
P
<
0.05)
and promoted cell apoptosis (
P
<
0.05).After hsa_circ_0000711 interference
the protein expression levels of Cyclin D1
CDK4 and Bcl-2 were decreased notably (
P
<
0.05)
but that of cleaved Caspase 3 was increased (
P
<
0.05).
Conclusion
2
Hsa_circ_0000711 may affect HCC cell proliferation by regulating cell cycle and cell apoptosis.
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