G蛋白偶联受体相关分选蛋白1在非小细胞肺癌中表达的临床意义及其与顺铂化疗抵抗的关系

Clinical significance of G-protein coupled receptor-associated sorting protein 1 expression in non-small cell lung cancer and its relationship with cisplatin chemotherapy resistance

目的探究G蛋白偶联受体相关分选蛋白1(GPRASP1)的表达与非小细胞肺癌(NSCLC)细胞的顺铂(DDP)敏感性的关系。方法收集癌旁组织、原发NSCLC组织及复发NSCLC组织,用荧光定量聚合酶链反应(RT-PCR)法检测GPRASP1和腺苷三磷酸结合盒式亚家族A成员5(ABCA5)mRNA的表达情况。空白组和过表达组分别用慢病毒转染空白载体和GPRASP1过表达载体于A549细胞中;对照组和抵抗组分别用慢病毒转染对照小干扰RNA(si-control)于A549细胞及DDP耐药细胞中;干扰组用慢病毒转染GPRASP1小干扰RNA(si-GPRASP1)于DDP耐药细胞中。用CCK-8实验法检测细胞对DDP的半抑制浓度(IC50),用RT-PCR法检测各组细胞中GPRASP1与ABCA5的表达情况。结果癌旁组织、NSCLC组织与复发NSCLC组织中GPRASP1 mRNA表达量分别为(1.27±0.16)×10^(-2)、(2.68±0.30)×10^(-2)和(3.89±0.72)×10^(-2),ABCA5 mRNA表达量分别为(1.03±0.13)×10^(-1)、(1.85±0.21)×10^(-1)和(2.67±0.61)×10^(-1);复发NSCLC组织中GPRASP1和ABCA5 mRNA表达量均显著高于癌旁组织及NSCLC组织,差异均有统计学意义(均P<0.01)。空白组和过表达组的GPRASP1 mRNA表达量分别为1.00±0.16和5.98±0.66,ABCA5 mRNA表达量分别为1.00±0.09和2.81±0.33,DDP IC50分别为(2.10±0.13)和(3.50±0.11)μmol·L^(-1),差异均有统计学意义(均P<0.01)。对照组、抵抗组和干扰组的GPRASP1 mRNA表达量分别为1.00±0.11、3.37±0.31和0.81±0.13,ABCA5 mRNA表达量分别为1.00±0.10、4.06±0.43和2.30±0.25,DDP IC50分别为(2.08±0.13)、(7.87±0.61)和(5.77±0.55)μmol·L^(-1);抵抗组的上述指标与对照组和干扰组比较,差异均有统计学意义(均P<0.01)。结论GPRASP1可显著促进ABCA5的表达,并介导NSCLC DDP抵抗。

Objective To explore the relationship between the expression of G-protein coupled receptor associated sorting protein 1(CPRASP1)and the cisplatin(DDP)sensitivity of non-small cell lung cancer(NSCLC)cells.Methods Normal paracancerous tissues,primary NSCLC tissues and recurrent NSCLC tissues after chemotherapy were collected,and the mRNA expression levels of GPRASP1 and adenosine triphosphate binding cassette subfamily A member 5(ABCA5)were detected by quantitative real-time polymerase chain reaction(RT-PCR).Blank and overexpression groups were transfected with the blank plasmid and GPRASP1 overexpression plasmid in A549 cells by lentivirus transfection technology,respectively.Control and resistance groups were transfected by the small interference control(si-control)plasmid in A549 and DDP resistance cells by lentivirus transfection technology,respectively.Interference group was transfected by small interference CPRASP1(si-GPRASPI)in DDP resistance cells.CCK-8 assay was used to detect the 50%inhibition concentration(ICso)of DDP,and RT-PCR was used to detect the mRNA expression levels of GPRASP1 and ABCA5.Results The expression levels of GPRASP1 mRNA in normal adjacent tissues,NSCLC tissues and recurrent NSCLC tissues were(1.27±0.16)×10^(-2),(2.68±0.30)×10^(-2)and(3.89±0.72)×10^(-2);the expression levels of ABCA5 mRNA were(1.03±0.13)×10^(-1),(1.85±0.21)×10^(-1)and(2.67±0.61)×10^(-1);the expression levels of GPRASP1 and ABCA5 mRNA in recurrent NSCLC tissues were significantly higher than those in normal adjacent tissues and NSCLC tissues,the differences were statistically significant(all P<0.O1).The expression levels of CPRASPI mRNA in blank and overexpression groups were 1.00±0.16 and 5.98±0.66;the expression levels of ABCA5 mRNA were 1.00±0.09 and 2.81±0.33;the DDP ICso were(2.10±0.13)and(3.50±0.11)μmol·L^(-1);the differences were statistically significant(all P<0.01).The expression levels of CPRASP1 mRNA in the control,resistance and interference groups were 1.00±0.11,3.37±0.31 and 0.81±0.13;the expression levels of ABCA5 mRNA were 1.00±0.10,4.06±0.43 and 2.30±0.25;the DDP ICso were(2.08±0.13),(7.87±0.61)and(5.77±0.55)μmol·L^(-1).Compared with the control and interference groups,the dfferences of above indexes in resistance group were statistically significant(all P<O.01).Conclusion GPRASP1 can significantly promote the expression of ABCA5 and mediate the DDP resistance of NSCLC.