长链非编码RNA SLC25A25-AS1对胃癌细胞及对5-氟尿嘧啶耐药的作用
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摘要
目的观察长链非编码RNA SLC25A25-AS1对胃癌(GC)细胞增殖、凋亡能力以及对5-氟尿嘧啶(5-Fu)化疗耐药的影响,并探讨其机制。方法采用实时荧光聚合酶链反应法(RT-PCR)检测正常胃黏膜(GSE-1)和GC细胞(SGC-7901,BGC-823)中SLC25A25-AS1和β-catenin的表达水平;在胃癌细胞SGC-7901和BGC-823中过表达SLC25A25-AS1后,通过RT-PCR检测β-catenin的表达水平;在胃癌细胞SGC-7901细胞中过表达SLC25A25-AS1后,再过表达β-catenin,应用CCK-8法检测转染后各组吸光值以评价增殖率,应用Annexin VAPC单染色流式细胞术检测各组转染48 h后的细胞凋亡率,在培养基中加入不同浓度的5-Fu检测各组细胞存活率。结果与正常胃黏膜细胞相比, GC细胞中SLC25A25-AS1低表达,β-catenin过表达(P<0.05);过表达SLC25A25-AS1后胃癌细胞β-catenin表达下调(P<0.05);胃癌SGC-7901细胞过表达SLC25A25-AS1后细胞增殖转移能力明显减弱(P<0.05),凋亡增加(P<0.05),化疗耐药减弱(P<0.05);而过表达β-catenin后恶性表型明显恢复(P<0.05)。结论 SLC25A25-AS1调节胃癌细胞增殖、凋亡和化疗耐药,参与GC的发生发展,与抑制β-catenin的表达相关。
Objective To study the role and mechanism of lncRNA SLC25 A25-AS1 in the proliferation,apoptosis and 5-Fu chemotherapy-resistant of gastric cancer(GC) cells.Methods The expression of β-catenin and SLC25 A25-AS1 in normal gastric mucosa cells(GSE-1) and GC cells(SGC-7901 and BGC-823) was detected by real-time fluorescent quantitative PCR. The proliferation of GC cells after overexpression of SLC25 A25-AS1 and overexpressed β-catenin was examined by CCK-8. Annexin V-APC staining was used to detect the apoptotic rates and viability of the cells.Results SLC25 A25-AS1 was weak-expressed in GC cells and the expression levels of β-catenin were significantly increased in GC cells than in normal gastric mucosa cells(P<0.05). Transfection with SLC25 A25-AS1 vector downregulated β-catenin expression(P<0.05). The abilities of proliferation, apoptosis and 5-Fu chemotherapy-resistant were significantly decreased after overexpression of SLC25 A25-AS1 in GC cells(P<0.05), which was reversed by the overexpression of β-catenin. Conclusion SLC25 A25-AS1 is involved in the occurrence and development of GC, its regulation effect on the proliferation, apoptosis and 5-Fu chemotherapy-resistant in GC cells is related to down-regulating the expression of β-catenin.
Objective To study the role and mechanism of lncRNA SLC25 A25-AS1 in the proliferation,apoptosis and 5-Fu chemotherapy-resistant of gastric cancer(GC) cells.Methods The expression of β-catenin and SLC25 A25-AS1 in normal gastric mucosa cells(GSE-1) and GC cells(SGC-7901 and BGC-823) was detected by real-time fluorescent quantitative PCR. The proliferation of GC cells after overexpression of SLC25 A25-AS1 and overexpressed β-catenin was examined by CCK-8. Annexin V-APC staining was used to detect the apoptotic rates and viability of the cells.Results SLC25 A25-AS1 was weak-expressed in GC cells and the expression levels of β-catenin were significantly increased in GC cells than in normal gastric mucosa cells(P<0.05). Transfection with SLC25 A25-AS1 vector downregulated β-catenin expression(P<0.05). The abilities of proliferation, apoptosis and 5-Fu chemotherapy-resistant were significantly decreased after overexpression of SLC25 A25-AS1 in GC cells(P<0.05), which was reversed by the overexpression of β-catenin. Conclusion SLC25 A25-AS1 is involved in the occurrence and development of GC, its regulation effect on the proliferation, apoptosis and 5-Fu chemotherapy-resistant in GC cells is related to down-regulating the expression of β-catenin.
引文
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