Effect and mechanism of long non-coding RNA MMP12-001 on the biological behavior of gallbladder cancer cells
Objective To investigate the expression of long non-coding RNA(lncRNA)MMP12-001 in gallbladder cancer(GBC)cells and its impact on the biological characteristics of GBC.Methods The expression of lncRNA MMP12-001 in GBC cancer tissues,adjacent tissues,GBC cell lines,and gallbladder epithelial cells was verified using qRT-PCR.The subcellular localization of lncRNA MMP12-001 was determined using RNA nuclear-cytoplasmic separation experiments.shRNA and overexpression lentiviral vectors specifically targeting lncRNA MMP12-001 were constructed and transfected into multiple GBC cell lines.The changes in cell proliferation ability were analyzed by cell survival rate detection and colony formation experiments.The changes in cell migration and invasion ability were observed using Transwell experiments.The interaction proteins of lncRNA MMP12-001 in GBC were initially screened using RNA pull down experiments combined with protein profiling and RNA binding protein immunoprecipitation(RIP).Results qRT-PCR verification showed that the expression level of lncRNA MMP12-001 in cancer tissues and GBC cell lines was significantly higher than that in adjacent tissues and gallbladder epithelial cells.RNA nuclear-cytoplasmic separation experiments showed that lncRNA MMP12-001 was mainly located in the cytoplasm.Overexpression of lncRNA MMP12-001 significantly promoted the proliferation,colony formation,migration,and invasion of GBC cells,with statistically significant differences(all P＜0.05).Silencing the expression of lncRNA MMP12-001 resulted in the opposite effect(all P＜0.05).RNA pull down combined with protein profiling identification showed that lncRNA MMP12-001 could directly bind to Sirtuin 3(SIRT3)in GBC.SIRT3 RIP experiments also found that SIRT3 IP could enrich lncRNA MMP12-001.Conclusion lncRNA MMP12-001 can promote the proliferation,invasion,and migration of GBC cells.
Long non-coding RNA MMP 12-001Gallbladder cancerProliferationInvasionMetastasis
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