Targeting Pyk2 gene on the proliferation, invasion and migration induction of hepatocelluar cancer Hep3B cells

Abstract

Objective To investigate the influence of proline-rich tyrosine kinase 2 (Pyk2) gene RNA interference on proliferation, invasion and migration of Hep3B hepatocellular carcinoma cells. Methods The Pyk2 gene RNA interference vector was transfected in Hep3B hepatocellular carcinoma cells by lipofectamine. The Hep3B cells divided into three groups: siRNA group (the vector with Pyk2 RNAi gene was transfected), negative control group (the vector without Pyk2 RNAi gene was transfected), and blank control group (no vectors was transfected). Pyk2 mRNA and protein were detected using reverse transcription reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. The biological behavior including cell proliferation, invasion and migration were detected by 3-(4, 5-dimethyl-2-thiazoly)-2, 5-diphenyl-2H-tetrazolium bromide (MTT), transwell and wound healing assay, respectively. Results The expression of Pyk2 mRNA of Hep3B cell line in siRNA group (0.16±0.03) was significantly decreased than those in negative group (0.74±0.13) and blank control group (0.77±0.16), with statistically significant differences (t=51.46, P=0.000; t=53.21, P=0.000). The expression of Pyk2 protein of Hep3B cell line in siRNA group (0.24±0.06) was significantly decreased than those in negative group (0.83±0.05) and blank control group (0.91±0.06), with statisti-cally significant differences (t=57.29, P=0.000; t=68.53, P=0.000). The cell proliferation inhibition rate at 48 hours in siRNA group (26.17%±0.28%) was significantly raised than those in negative group (9.28%±0.22%) and blank control group (6.47%±0.31%), with statistically significant differences (t=31.45, P=0.004; t=34.64, P=0.002). The number of transmembrane cells in siRNA group (32.5±8.5)/10 HP was significantly declined than those in negative group (98.4±12.3)/10 HP and blank control group (112.6±11.3)/10 HP, with statistically significant differences (t=95.64, P=0.000; t=105.17, P=0.000). The wound healing assay in siRNA group (28.17%±1.46%) was significantly lower than those in negative group (77.38%±2.24%) and blank control group (79.41%±3.17%), with statistically significant (t=85.86, P=0.000; t=89.37, P=0.000). Conclusion Pyk2 gene involves the proliferation, invasion and migration of Hep3B cells, which has close correction with development and metastasis of hepatocellular carcinoma. Pyk2 gene is very helpful to become a molecular target for the diagnosis and treatment of hepatocellular carcinoma. Key words: Liver neoplasms; Cell proliferation; Neoplasm invasiveness; RNA interference; Proline-rich tyrosine kinase 2