Effects Of GM130 Gene Suppression On Proliferation, Invasion And Migration Of Human Gastric Carcinoma Cell

Objective:To explore the expression of GM30 in the differentiation of gastric carcinoma cell lines and investigate the effect of small interference RNA-mediated silcencing of the GM130 gene on cell proliferation, invasion and migration of human gastric carcinoma MKN-45 cells in vitro.Methods:Three different differentiated gastric carcinoma cell lines (MKN-28, SGC-7901 and MKN-45) were cultured in vitro. Western blot and RT-PCR analyses were carried out in MKN-45 cells and SGC-7901 cells, which had high GM130 expression levels. The designed and chemically synthesized siRNA targeting the GM130 gene was transfected into MKN-45 cells. The effect of GM130 knockdown on MKN-45 cells proliferation was determined by MTT assay, cells migration was analyzed by Transwell migration assay, the invasion of cells was determined by Matrigel invasion assay. The expression of MMP-2 and -9 was also detected at the protein level.Results:MKN-45 and SGC-7901 cells showed the higher expression of GM130 at the protein and mRNA levels, so Western blot was carried out in MKN-45cells, which had invasive and metastatic potential. The GM130-siRNA-519 significantly decreased GM130 protein level compared to two other sequences of the siRNA targeting GM130 and scrambled. The distinct decrease of GM130 mRNA expression was further confirmed by quantative Real-time PCR in MKN-45 cells (P< 0.05). Compared with the control group, the inhibition of GM130 expression by siRNA which had lower levels of invasion and migration capacity suppressed the proliferation in MKN-45 cells. The cells transfected with GM130 siRNA showed significantly suppressed migration ability with less cells moved from the upper chamber into the lower, and obviously decreased expressed of MMP-2 and -9 at the protein level (P< 0.05)Conclusion:SiRNA-mediated silencing of the GM130 gene could significantly inhibited cell proliferation, migration and invasion in human gastric cancer cell line MKN-45.