Role and mechanism of oncogene SKA3 in proliferation regulation of hepatocellular carcinoma cells
10.3877/cma.j.issn.2095-3232.2018.03.016
- VernacularTitle:癌基因SKA3在肝癌细胞增殖调控中的作用及机制
- Author:
Jianxin TANG
1
;
Jinjun LI
;
Ziming LIANG
;
Nan JIANG
Author Information
1. 中山大学附属第三医院肝脏外科肝移植中心
- Keywords:
Carcinoma;
hepatocellular;
SKA3;
Cell proliferation;
Cell cycle
- From:
Chinese Journal of Hepatic Surgery（Electronic Edition）
2018;7(3):236-240
- CountryChina
- Language:Chinese
-
Abstract:
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 byt 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.
