Effects of knocking out Bcl-2 gene on proliferation and apoptosis of human pancreatic cancer cells SW1990
10.3760/cma.j.issn.1674-1935.2015.04.005
- VernacularTitle:Bcl-2基因敲除对人胰腺癌细胞增殖及凋亡的影响
- Author:
Li WEI
;
Haiwen ZHANG
;
Qianqian TU
;
Bin LIU
;
Hongjian CAI
;
Chunliang SUN
;
Haitao CHEN
- Publication Type:Journal Article
- Keywords:
Pancreatic neoplasms;
Genes,bcl-2;
CRISPR-Cas9;
Cell proliferation;
Apoptosis
- From:
Chinese Journal of Pancreatology
2015;15(4):237-241
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of Bcl-2 gene expression on the proliferation and apoptosis of human pancreatic cancer SW1990 cells.Methods Bcl-2 short guide RNA (Bcl-2-sgRNA) was designed and synthesized,and it was combined with CRISPR-Cas 9.After confirmation by gene sequencing,it was transfected into human pancreatic cancer cell line SW1990,then the cells with stable Bcl-2 gene knock-out were selected,and wild type SW1990 cells were used as control.The cell growth curve was determined by CCK-8 method.The number of clone formation was measured.Flow cytometry was used to measure cell cycle and apoptosis.Results Human pancreatic cancer cell line SW1990 with Bcl-2 gene knock-out was successful constructed.Compared with wild type SW1990 cells,the growth of SW1990 cells with Bcl-2 gene knock-out was inhibited,the number of clone formation was significantly decreased [(160.7 ± 10.0) vs (285.3 ± 14.2)],the proportion of G1 cells was significandy increased [(84.51 ± 0.97) ％ vs (57.49 ± 1.08) ％],the proportion of S phase cells significantly decreased [(12.82 ± 0.99) ％ vs (27.56 ± 1.65) ％],and apoptosis rate was remarkably increased [(12.67 ± 0.59) ％ vs (0.37 ± 0.35) ％],and the difference between the two groups was statistically significant (P ＜ 0.01).Conclusions Knock-out of Bcl-2 gene can inhibit the growth of human pancreatic cancer cell line SW1990,decrease the ability of clone formation,block the cell in G1 phase and greatly increase cell apoptosis rate.
