摘要
Abstract
Objective To explore the mechanism of long non-coding RNA prostate cancer-associated transcript 19(LncRNA PCAT19)knockdown on oxaliplatin(OXA)sensitivity of human gastric cancer cells by regulating microRNA(miR)-200c-3p mediated ataxia-telangiectasia mutated proteins(ATM)and Rad3-related kinase-related kinase(ATR)/checkpoint kinase1(CHK1)signaling pathway.Methods Human gastric adenocarcinoma cells(AGS)were randomly divided into OXA group(8 μg·L-1 OXA),OXA+si-NC+inh-NC group(si-NC and inh-NC co-transfected with+8 μg·L-1 OXA),OXA+si-PCAT19+inh-NC group(si-PCAT19 and inh-NC co-transfected+8 μg·L-1 OXA),OXA+si-PCAT19+miR-200c-3p inh group(si-PCAT19 and miR-200c-3p inh co-transfected+8 μg·L-1 OXA).Cell proliferation in each group was detected by 5-acetylidene-2′-deoxyuridine assay;cell apoptosis was detected by in situ terminal transferase labeling technique;cell migration was detected by cell scratch assay;the expression levels of phosphorylated histone H2A.X(γH2A.X)and radiation-sensitive protein 51(RAD51)were detected by immunofluorescence.Results In OXA group,OXA+si-NC+inh-NC group,OXA+si-PCAT19+inh-NC group,OXA+si-PCAT19+miR-200c-3p group,the cell proliferation rates were(58.83±6.52)％,(59.42±6.86)％,(45.59±6.09)％and(55.71±7.69)％,respectively;the apoptosis rates were(20.06±2.44)％,(20.32±2.66)％,(31.74±4.43)％and(24.60±3.29)％,respectively;the migration rates were(54.50±6.14)％,(56.11±6.72)％,(31.51±4.13)％and(48.59±7.26)％,respectively;the relative fluorescence intensities of γH2A.X were 1.00±0.08,0.97±0.12,4.84±0.82 and 2.11±0.41,respectively;the relative fluorescence intensities of RAD51 were 1.00±0.06,1.08±0.15,0.22±0.04 and 0.55±0.08,respectively.There were no significant differences between OXA+si-NC+inh-NC group and OXA group(all P＞0.05).The above indexes of OXA+si-PCAT19+inh-NC group were compared with those of OXA+si-NC+inh-NC group,the above indexes in OXA+si-PCAT19+miR-200c-3p inh group were compared with those of OXA+si-PCAT19+inh-NC group,the differences were statistically significant(all P＜0.05).Conclusion Knockdown of LncRNA PCAT19 can affect the proliferation,migration,apoptosis and deoxyribonucleic acid repair of gastric cancer cells by targeting and negatively regulating miR-200c-3p,thereby enhancing the sensitivity of gastric cancer cells to OXA,which may be related to the ATR/CHK1 signaling pathway.关键词
奥沙利铂/胃癌/耐药性/DNA修复/长链非编码RNAKey words
oxaliplatin/stomach neoplasms/drug resistance/DNA repair/long non-coding RNA分类
医药卫生
