人核糖体蛋白S19在非小细胞肺癌中的表达及与顺铂耐药的关系

Expression of human ribosomal protein S19 in non-small cell lung cancer and its association with cisplatin resistance

目的分析人核糖体蛋白S19(RPS19)在非小细胞肺癌(non-small cell lung cancer,NSCLC)中的表达,及与顺铂(DDP)耐药的关系。方法根据DDP化疗的疗效,将50例NSCLC患者分为有效组与无效组,采用实时荧光定量PCR(RT-qPCR)法和蛋白免疫印迹(WB)法检测患者癌组织与癌旁组织中RPS19表达情况。将si-RPS19质粒、si-阴性对照(NC)序列分别转染至人NSLCL耐顺铂株A549/DDP细胞,并设为si-RPS19组、si-NC组、空白对照组,采用RT-qPCR、WB法检测各组RPS19的表达,CCK-8法和流式细胞术分别检测DDP对细胞的半抑制浓度(IC_(50))和细胞凋亡率,采用WB检测凋亡相关蛋白Bax、Bcl-2、caspase-3、caspase-9的表达。结果NSCLC患者癌组织中RPS19 mRNA及蛋白相对表达量(0.61±0.11、0.53±0.09)均较癌旁组织(0.38±0.06、0.21±0.05)升高,差异均有统计学意义(t=12.980、21.978,均P<0.001)。各组A549/DDP细胞的存活率随DDP浓度的增加而下降,凋亡率随DDP浓度的增加而升高,差异均有统计学意义(P<0.05);其中,si-RPS19组A549/DDP细胞存活率最低,凋亡率最高。与空白对照组、si-NC组比较,si-RPS19组的Bax、caspase-3、caspase-9蛋白相对表达量升高,Bcl-2蛋白相对表达量降低;差异均有统计学意义(P<0.05)。结论RPS19在NSCLC中呈高表达,敲低RPS19可抑制A549/DDP细胞的活性,促进癌细胞凋亡,增强癌细胞对DDP的敏感性。

Objective To analyse the expression of human ribosomal protein S19(RPS19)in non-small cell lung cancer(NSCLC)and its association with cisplatin(DDP)resistance.Methods According to the efficacy of DDP chemotherapy,50 NSCLC patients were divided into effective group and ineffective group,real-time fluorescence quantitative PCR(RT-qPCR)and protein Western blot(WB)were used to detect the expression of RPS19 in cancer tissues and adjacent tissues of patients.si-RPS19 plasmid and si-negative control(NC)sequence were transfected into human NSLCL cisplatin-resistant A549/DDP cells and divided into si-RPS19 group,si-NC group and blank control group,respectively,RT-qPCR and WB methods were used to detect the expression of RPS19 in each group,CCK-8 method and flow cytometry were used to detect the semi-inhibitory concentration(IC_(50))and apoptosis rate of DDP cells,respectively,and WB was used to detect the expression of apoptosis-related proteins Bax,Bcl-2,caspase-3 and caspase-9.Results The relative expression levels of RPS19 mRNA and protein in cancer tissues of NSCLC patients(0.61±0.11,0.53±0.09)were higher than those in adjacent tissues(0.38±0.06,0.21±0.05),with statistical significance(t=12.980,21.978,P<0.001).The survival rate of A549/DDP cells exhibited a significant decrease with increasing DDP concentration in all experimental groups,while the apoptosis rate demonstrated a statistically significant increase(P<0.05).Notably,the si-RPS19 group displayed the lowest survival rate and the highest apoptosis rate among A549/DDP cells.Compared with the blank control group and the si-NC group,the relative expression of Bax,caspase-3 and caspase-9 protein was increased in the si-RPS19 group,while the relative expression of Bcl-2 protein was decreased;the differences were statistically significant(P<0.05).Conclusions RPS19 is highly expressed in NSCLC and knockdown of RPS19 can inhibit the activity of A549/DDP cells,promote apoptosis of cancer cells and increase the sensitivity of cancer cells to DDP.