Effects of knocking out Bcl-2 gene on proliferation and apoptosis of human pancreatic cancer cells SW1990

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 significantly 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. Key words: Pancreatic neoplasms; Genes, bcl-2; CRISPR-Cas9; Cell proliferation; Apoptosis