Effect of silencing phosphatidylinositol 3-kinase regulatory subunit gamma gene by small interfering RNA on proliferation and migration capability of gallbladder carcinoma cells

Abstract

Objective To observe the effect of small interfering RNA (siRNA) silencing phosphatidylinositol 3-kinase regulatory subunit gamma (PIK3R3) gene on the proliferation and migration of gallbladder cancer cells NOZ and GBC-SD. Methods Small interfering RNA (si-PIK3R3) against PIK3R3 was constructed and real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting were used to detect PIK3R3 mRNA and protein in transfected gallbladder cancer cells NOZ and GBC-SD. The expression of cell counting kit-8 (CCK-8) proliferation assay was used to detect the proliferation of transfected 0-5 d cells. Transwell chamber assay was used to detect the migration of gallbladder cancer cells 48h after transfection. Results The si-PIK3R3 transfection group and the si-NC negative control group were successfully transfected into the NOZ and GBC-SD of gallbladder cancer cells. The expression of PIK3R3 mRNA (0.119±0.447) in the NOZ cell si-PIK3R3 group was significantly lower than that of NC group (1.000±0.398) (t=165.100, P<0.05), the PIK3R3 mRNA (0.085±0.593) in the si-PIK3R3 group of GBC-SD cells was significantly lower than that in the si-NC group (1.000±0.669) (t=500.200, P<0.05); the expression of PIK3R3 protein in the transfected group was significantly lower than that in the negative control group. The results of CCK-8 in NOZ cells showed that the absorbance values of 0-5 d si-PIK3R3 transfection group were 0.185±0.007, 0.163±0.013, 0.229±0.019, 0.240±0.017, 0.273±0.012, 0.350±0.069 (t=11.960, P<0.05); The results of 48 h Transwell showed that the number of cells migrated in the si-PIK3R3 transfection group [(69.0±4.5) cells] was significantly lower than that in the si-NC group (t=25.120, P<0.05). The results of CCK-8 in GBC-SD cells showed that the absorbance values of the 0-5 d si-PIK3R3 transfection group were 0.153±0.003, 0.158±0.002, 0.201±0.010, 0.240±0.012, 0.297±0.018, 0.483±0.024 (t=14.160, P<0.05); 48 h Transwell results showed that the number of cells migrated in the si-PIK3R3 transfection group [(91.0±3.5) cells] was significantly lower than that in the si-NC group (t=25.390, P<0.05). Conclusion si-PIK3R3 can inhibit the proliferation and migration of human gallbladder cancer cells, suggesting that PIK3R3 plays an important role in the development of gallbladder carcinoma. Key words: Gallbladder cancer; Small interfering RNA; Phosphatidylinositol 3-kinase regulatory subunit gamma