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中华肝脏外科手术学电子杂志 ›› 2018, Vol. 07 ›› Issue (03) : 236 -240. doi: 10.3877/cma.j.issn.2095-3232.2018.03.016

所属专题: 文献

基础研究

癌基因SKA3在肝癌细胞增殖调控中的作用及机制
唐建信1, 李进军1, 梁梓明1, 姜楠1,()   
  1. 1. 510630 广州,中山大学附属第三医院肝脏外科 肝移植中心
  • 收稿日期:2018-03-16 出版日期:2018-06-10
  • 通信作者: 姜楠
  • 基金资助:
    国家自然科学基金(81572368); 广东省自然科学基金(2016A030313278)

Role and mechanism of oncogene SKA3 in proliferation regulation of hepatocellular carcinoma cells

Jianxin Tang1, Jinjun Li1, Ziming Liang1, Nan Jiang1,()   

  1. 1. Liver Transplantation Center, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China
  • Received:2018-03-16 Published:2018-06-10
  • Corresponding author: Nan Jiang
  • About author:
    Corresponding author: Jiang Nan, Email:
引用本文:

唐建信, 李进军, 梁梓明, 姜楠. 癌基因SKA3在肝癌细胞增殖调控中的作用及机制[J/OL]. 中华肝脏外科手术学电子杂志, 2018, 07(03): 236-240.

Jianxin Tang, Jinjun Li, Ziming Liang, Nan Jiang. Role and mechanism of oncogene SKA3 in proliferation regulation of hepatocellular carcinoma cells[J/OL]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2018, 07(03): 236-240.

目的

探讨癌基因纺锤体与着丝粒相关蛋白3(SKA3)在肝癌细胞增殖调控中的作用及机制。

方法

采用Western blot和RT-PCR检测肝癌组织和细胞SKA3表达水平。采用慢病毒感染构建SKA3过表达组(Huh7-SKA3)和对照组(Huh7-Vector),构建SKA3干扰组(Huh7-shSKA3)和对照组(Huh7-Scramble)细胞模型。采用MTT法和细胞克隆形成实验检测细胞增殖能力,流式细胞仪检测细胞周期,Western blot和RT-PCR检测细胞周期相关蛋白的表达。实验数据比较采用t检验。

结果

肝癌组织和Huh7肝癌细胞SKA3蛋白表达明显增强,mRNA表达量分别为15.46±2.10、9.26±0.80,明显高于正常肝组织和LO2正常肝细胞的1.48±0.17、1.11±0.07(t=6.58,10.08;P<0.05)。细胞生长曲线显示过表达组细胞的增殖能力明显强于对照组,干扰组细胞的增殖能力低于其对照组。过表达组细胞克隆数为(259±20)个,明显高于对照组的(97±11)个(t=9.12,P<0.05),干扰组细胞克隆数为(11±2)个,明显少于对照组的(97±11)个(t=-8.13,P<0.05)。过表达组G2/M期和S期细胞比率分别为(5.5±0.6)%和(41.9±3.6)%,对照组相应为(15.8±1.5)%和(31.9±3.3)%,差异有统计学意义(t=-10.89,6.39;P<0.05),而干扰组G1/G0期和S期细胞比率分别为(71.7±6.1)%、(20.2±2.9)%,对照组相应为(53.4±5.4)%、(31.3±4.2)%,差异有统计学意义(t=8.59,-5.17;P<0.05)。

结论

SKA3在肝癌中高表达,可促进肝癌细胞增殖,可能与促进细胞周期G1期进入S期有关。

Objective

To investigate the role and mechanism of oncogene spindle and kinetochore associated complex subunit 3 (SKA3) in the proliferation regulation of hepatocellular carcinoma (HCC) cells.

Methods

The expression of SKA3 in HCC tissues and cells was detected by Western blot and RT-PCR. SKA3 overexpression group (Huh7-SKA3) and control group (Huh7-Vector) were constructed using lentivirus transfection. Cell models of SKA3 interference group (Huh7-shSKA3) and control group (Huh7-Scramble) were constructed as well. The ability of cell proliferation was detected by MTT assay and cell clony formation assay. The cell cycle was detected by flow cytometry, and the expression of cyclin related proteins was detected by Western blot and RT-PCR. The experimental data were compared by t test.

Results

The expression of SKA3 protein in HCC tissues and Huh7 HCC cells increased significantly, and the expression level of mRNA was respectively 15.46±2.10 and 9.26±0.80, significantly higher than 1.48±0.17 and 1.11±0.07 of normal liver tissues and normal liver cells LO2 (t=6.58, 10.08; P<0.05). The cell growth curve showed that the proliferation ability of cells in overexpression group was significantly better than those in control group, and the proliferation ability of cells in interference group was lower than those in control group. The number of cell clones in the overexpression group was 259±20, significantly higher than 97±11 in control group (t=9.12, P<0.05), and the number of cell clones in interference group was 11±2, significantly lower than 97±11 in control group (t=-8.13, P<0.05). The ratio of phase G2/M and phase S in overexpression group was respectively (5.5±0.6)% and (41.9±3.6)%, and was correspondingly (15.8±1.5)% and (31.9±3.3)% in control group, and significant differences were observed (t=-10.89, 6.39; P<0.05). The ratio of phase G1/G0 and phase S in interference group was respectively (71.7±6.1)% and (20.2±2.9)%, and was correspondingly (53.4±5.4)% and (31.3±4.2)% in control group, and significant differences were observed (t=8.59, -5.17; P<0.05).

Conclusions

SKA3 is highly expressed in HCC and can promote the proliferation of HCC cells, which may be related to its promotion of cell cycle from phase G1 to phase S.

图1 Western blot检测SKA3在肝癌组织和细胞中高表达
图2 MTT法检测肝癌细胞的生长曲线
图3 RT-PCR检测肝癌细胞周期相关蛋白表达
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