The Functional Analysis And Clinical Significance Of Methylase ASH2L And KDM4B In Pancreatic Cancer

ObjectiveBased on constructing histone methylation related enzymes virus library, we usedthe functional high throughput screening technology to seek out pancreatic cancer proliferationrelatedhistone methylation genes.MethodsUse of nonspecific lentivirus and cell lines, determine the infection condition and MTT conditions. Based on the high throughput lentiviral vector functional epigenome RNA library, to screen effective cell proliferation epigenome genes and small interference RNA in pancreatic cancer cell lines.ResultsThe best cell density is10%, useing the50ul virus infect cells96h later. Based on the high throughput lentiviral vector functional epigenome RNA library, to screen effective cell proliferation realted epigenome genes:PRMT8, ASH2L, PRDM9, JMJD8, KDM4B.ConclusionFunctional high throughput screening technology can screen out proliferation related epigenome genesin pancreatic cancer cells, and the potential role of genes in pancreatic cancer will be investigate in the part2. ObjectiveTo investigate the ASH2L and KDM4B in human pancreatic carcinoma cell lines and explore the potential role of them in pancreatic carcinoma.MethodsBased on the high throughput lentiviral vector functional epigenome RNA library, we screened effective cell proliferation epigenome genes andsmall interference RNA. Regulate genes expression andconduct the migration tests, FACS analysisand Western blot tests. Effect of the positive genes on the cell proliferation, invasiveness cell cycle distribution and apoptosis were investigated. The potential signaling pathways were detected by western blot.ResultsBased on the high throughput lentiviral vector functional epigenome RNA library, to screen effective cell proliferation epigenome genes andsmall interference RNA:ASH2L:0044-08D, KDM4B:0029-08G In vitro experiment, upregulation of ASH2L and KDM4B in pancreatic carcinoma cell line PANC-1reduce cell proliferation and invasiveness. FACS analysis showed ASH2Land KDM4B can block cell cycle in G2/M phase. Western blotindicatedASH2L and KDM4B influenced pancreatic cancer cell migration ability via the Src and FAK pathway.ConclusionASH2L and KDM4B gene inhibits cell proliferation and invasion ability, and it may be through the Akt, ERK,Src and FAK signal pathway influence pancreatic cancer cell proliferation andmigration ability. ObjectiveTo investigate the expression of KDM4B in pancreatic ductal adenocarcinomasamples and analyze their correlation with patients’s clinicopathological characteristics.MethodsWe collected the paraffin-embedded samples,as well as medical records and follow-up data of50patients with pancreatic cancer, who were treated in Zhongshan Hospital interventional department in2007～2012, The KDM4B expressions in pancreatic cancer tissues were evaluated according to the immunohistochemical staining area and intensity. Statistical analysis KDM4B expression level and the correlation between the patients’clinical and pathological characteristics.ResultsThe overall expression rate of KDM4B was54.0%in pancreatic cancer tissues, and was18.0%in the adjacent tissues.The KDM4B expression was no significantly associated with sex, age, tumor gradeand tumor location(P>0.05). TNM stage was related to the expression. In multivariate analysis, KDM4B was the independent factors of tumor progression.ConclusionsKDM4B was highly expressed in pancreatic cancer and correlated with tumor progression. ObjectiveDiscuss the relationship of KDM4B with gemcitabine drug sensitivity.MethodsPANC-1cells stably knockdowning KDM4B were screen by puromycin and verified by qPCR. The concentration of gemcitabine to treat PANC-1cells was determined by MTT assay to make cell viability decrease by30%or so. The effect of KDM4B suppression on chemosensitivity to gemcitabine ofPANC-1was observed through viability of cells before and after gemcitabine treatmentResultsqPCR results showed that KDM4B expression was suppressed by%in PANC-1cells stably knockdowning KDM4B. Cell viability of the stable cell line was not significantly altered before and after treatment of100nM gemcitabine.ConclusionSilence KDM4B genes cannot increase gemcitabine in pancreatic cancer cell proliferation inhibition, not enough to improve pancreatic cancer to gemcitabine chemotherapy sensitivity.