胃癌患者癌组织及血清中lncRNA-GC1的表达及其对化疗耐药性的影响

Expression of lncRNA-GC1 in cancer tissue and serum of patients with gastric cancer and its effect on chemotherapy resistance

目的探讨长链非编码RNA-GC1(lncRNA-GC1)在胃癌筛查中的价值,以及对胃癌患者化疗耐药性的影响。方法选取2019年1月至2021年1月于该院行手术治疗的86例胃癌患者作为研究对象,另纳入同期进行体检的86例健康体检者作为健康对照组。收集胃癌患者的血液及组织标本,收集健康对照者的血液标本。采用实时荧光定量PCR测定组织和血清lncRNA-GC1表达水平。选取胃癌细胞MKN-45进行培养,建立稳定转染胃癌细胞系MKN-45 GC1,对照细胞为MKN-45 Vecc。应用噻唑蓝(MTT)实验检测细胞活力,3-D细胞培养实验检测细胞增殖和生长能力。结果胃癌患者癌组织中lncRNA-GC1表达水平明显高于癌旁组织(1.45±0.16 vs.0.82±0.07,t=15.362,P<0.001);血清lncRNA-GC1表达水平明显高于健康对照组(0.78±0.13 vs.0.54±0.05,t=6.887,P<0.001)。胃癌患者癌组织中lncRNA-GC1表达水平与肿瘤最大径、神经侵犯、淋巴结转移有关(均P<0.05),胃癌患者血清中lncRNA-GC1表达水平与肿瘤最大径、淋巴结转移和临床分期有关(均P<0.05)。受试者工作特征(ROC)曲线分析结果显示,血清lncRNA-GC1水平诊断胃癌的曲线下面积为0.849[95%CI(0.790,0.907),P<0.001]。转染后的MKN-45 GC1细胞中lncRNA-GC1表达水平明显高于MKN-45 Vec细胞。MTT实验结果表明,MKN-45 GC1细胞的细胞活力较MKN-45 Vec细胞明显升高;加入顺铂后,MKN-45 GC1和MKN-45 Vec细胞的细胞活力均下降,但MKN-45 GC1细胞的细胞活力仍明显高于MKN-45 Vec细胞。3-D细胞培养实验显示,MKN-45 GC1细胞的增殖能力明显高于MKN-45 Vec细胞;加入6μg/mL顺铂后,两种细胞的增殖能力均受到抑制,但MKN-45 GC1细胞增殖能力明显高于MKN-45 Vec细胞。结论lncRNA-GC1在胃癌患者组织和血清中表达上调,可作为胃癌筛查及预后判断的分子标志物,且lncRNA-GC1可促进胃癌细胞MKN-45的增殖生长,并促进细胞对顺铂的耐药。

Objective To explore the value of long non-coding RNA-GC1(lncRNA-GC1)in gastric cancer screening and its effect on chemotherapy resistance in patients with gastric cancer.Methods A total of 86 patients with gastric cancer who underwent surgical treatment in the hospital from January 2019 to January 2021 were selected as the research subjects,and a total of 86 healthy people who underwent physical examination in this hospital during the same period were enrolled as the control group.Blood and tissue samples were collected from patients with gastric cancer,and blood samples from healthy controls were collected,as well.Real-time fluorescent quantitative PCR was used to determine the expression levels of lncRNA-GC1 in tissues and serum.The gastric cancer cell line MKN-45 was selected for culture.The stable transfected gastric cancer cell line MKN-45 GC1 was established,and the control cell was MKN-45 Vecc.Methyl thiazolyl tetrazolium(MTT)experiment was used to detect cell viability,and 3-D cell culture experiment was used to detect cell proliferation and growth ability.Results The expression level of lncRNA-GC1 in cancer tissues of patients withgastric cancer was significantly higher than that in adjacent tissues(1.45±0.16 vs.0.82±0.07,t=15.362,P<0.001).The expression level of serum lncRNA-GC1 of patients with gastric cancer was significantly higher than that of the healthy control group(0.78±0.13 vs.0.54±0.05,t=6.887,P<0.001).The expression level of lncRNA-GC1 in cancer tissue of patients with gastric cancer was related to the maximum diameter of tumor,nerve invasion,and lymph node metastasis(all P<0.05),and the expression level of lncRNA-GC1 in serum of patients with gastric cancer was related to the maximum diameter of tumor,lymph node metastasis and clinical stage(all P<0.05).Receiver operating characteristic(ROC)curve analysis results showed that the area under the curve for the diagnosis of gastric cancer by serum lncRNA-GC1 level was 0.849[95%CI(0.790,0.907),P<0.001].The expression level of lncRNA-GC1 in MKN-45 GC1 cells after transfection was significantly higher than that in MKN-45 Vec cells.The results of MTT assay showed that the viability of MKN-45 GC1 cells was significantly higher than that of MKN-45 Vec cells.After adding cisplatin,the viability of MKN-45 GC1 and MKN-45 Vec cells decreased,but the viability of MKN-45 GC1 cells was still significantly higher than that of MKN-45 Vec cells.3-D cell culture experiments showed that the proliferation ability of MKN-45 GC1 cells was significantly higher than that of MKN-45 Vec cells.After adding 6μg/mL cisplatin,the proliferation of both cells was inhibited,but the proliferation of MKN-45 GC1 cells was significantly higher than that of MKN-45 Vec cells.Conclusion The expression of lncRNA-GC1 is up-regulated in the tissues and serum of patients with gastric cancer,which can be used as a molecular marker for gastric cancer screening and prognostic judgment.And lncRNA-GC1 can promote the proliferation and growth of gastric cancer cells MKN-45,and promote the resistance of cells to cisplatin.