NOLC1基因在胃癌组织中的表达及其对顺铂耐药性的影响

Expression of NOLC1 gene in gastric cancer tissues and its effect on cisplatin resistance

目的:研究NOLC1在胃癌中的表达量以及与顺铂耐药的相关性。方法:使用肿瘤基因组图谱(TCGA)数据库对NOLC1在胃癌及正常胃组织表达水平进行评估,应用KaplanMeier生存分析预测NOLC1在胃癌患者中的预后价值。通过免疫组化检测临床胃癌及其癌旁正常标本中NOLC1的表达量。通过持续添加低浓度顺铂诱导构建顺铂耐药株细胞。使用免疫荧光和Westernblot检测耐药型和敏感型细胞中NOLC1表达水平。使用慢病毒构建NOLC1敲低的稳转株胃癌细胞。使用不同浓度的顺铂处理胃癌细胞24h后,通过CCK-8、克隆形成和流式细胞术检测NOLC1对胃癌对顺铂耐药的影响,使用Westernblot检测NOLC1敲低后顺铂诱导的细胞凋亡相关蛋白表达水平。结果:NOLC1基因在胃癌组织中高表达(P<0.01)并且其高表达预示不良预后(P<0.01)。NOLC1在对顺铂耐药的胃癌细胞株中高表达(P<0.01)。敲低NOLC1后,顺铂对胃癌细胞的杀伤能力和增殖抑制效果增强(P<0.05),并且细胞中Cl-Caspase3、Cl-PARP蛋白等凋亡相关蛋白表达上调(P<0.01)。结论:NOLC1蛋白在胃癌中高表达并可以促进胃癌对顺铂的耐受。

Objective:To investigate the expression of NOLC1 in gastric cancer and its correlation with cisplatin resistance.Methods:The Cancer Genome Atlas(TCGA)database was used to evaluate the expression level of NOLC1,and Kaplan-Meier survival analysis was used to predict the prognostic value of NOLC1 in gastric cancer patients.The expression of NOLC1 in gastric cancer tissues was detected by immunohistochemistry.The cisplatin-resistant cells were induced by continuous addition of cisplatin at low concentration.The expression of NOLC1 in drug-resistant and sensitive cells was detected by immunofluorescence and Western blot.A stable NOLC1 knockdown gastric cancer cell line was constructed by lentivirus.Gastric cancer cells were treated with different concentrations of cisplatin for 24 h.CCK-8 assay,colony formation assay,and flow cytometry were used to detect the effect of NOLC1 on cisplatin resistance of gastric cancer cells.Western blot was used to detect the expression level of apoptosis-related proteins induced by cisplatin after NOLC1 knockdown.Results:NOLC1 was highly expressed in gastric cancer tissues(P<0.05),which predicted poor prognosis(P<0.01).NOLC1 was highly expressed in cisplatin-resistant gastric cancer cell lines(P<0.01).After NOLC1 knockdown,cisplatin enhanced the killing ability and proliferation inhibition of gastric cancer cells(P<0.05),and the expression of apoptosis-related proteins such as Cl-Caspase3 and Cl-PARP was up-regulated(P<0.01).Conclusion:NOLC1 is highly expressed in gastric cancer and can promote the resistance of gastric cancer to cisplatin.